1

Replacement of yellow corn with mangrove seeds in diet

for in Bluespot mullet

Ibrahim E.H. Belal

Department of Aquatic Resources Development, College of Agricultural and Food Sciences,

King Faisal University, Al- Hassa, Saudi Arabia

Key words; Fish, Feed, Recirculating System, Mangrove Seeds, Yellow Corn Correspondence: Dr I. E. H. Belal Department of Aquatic Resources Development, College of Agriculture and Food Sciences, King Faisal University, PO Box 420, Al-Hassa 31982, Saudi Arabia ibelal@yahoo.com , Home (T. & Fax) – (966) 356-602109 Work (T.) - (966) 5800000/ 1412

2

Replacement of yellow corn with mangrove seeds in diet for in

Bluespot mullet1

Abstract

Four approximately isoenergetic isonitrogenous diets containing 0g

kg-1, 150 g kg-1, 300 g kg-1 and 510 g kg-1 mangrove seeds as a

replacement for dietary corn in bluespot mullet Valamugil seheli

(Valenciennes) commercial feed were fed to triplicate groups (100 fish

each) of fingerlings (0.5g) for ten weeks. The closed re-circulating system

consisted of 12 cubical tanks (2.25 m3 each). Fish were fed three times a

day to satiation for 12 weeks. Growth ranked the diets 300 > 150 > 0 >

500 g kg-1 mangrove seed substitution and in most cases differences in fish

weight or SGR were significant (P < 0.05). The proximate composition of

the fish bodies was affected (P < 0.05) by replacing dietary corn with

mangrove seeds in the test diets. As the level of the mangrove seed

incorporation increased, fish body moisture, ash, and protein were

1 This paper is written from research project # 1014 that was financially supported by the Deanship of Scientific Research, King Faisal University Al-Hassa, Eastern Province, Saudi Arabia.

3

increased and body fat reduced. Inclusion of 300g kg-1 mangrove seed

meal as a replacement for corn resulted in good performance.

4

Introduction

The gray mangrove plant Avicennia marina (Forsk.) Vierh. is an

annual salt-marsh tree that has great potential as a crop in arid regions. The

primary advantages of mangrove are that it can be irrigated entirely with sea

water, it is adaptable to arid conditions and it is highly productive (Gordon et

al. 1998) ). The average weight of mangrove seeds is 3.03 g, each shrub

produces an average of 580 seeds and calculated seed productivity is 11.6

ton hectare-1 year-1(Farah, 2002). Additionally, mangrove seeds are a good

source of iodine and provide approximatly 863 g kg-1 carbohydrate, 105 g

kg-1 crude protein, 9 g kg-1 crude fat, and 2.1 g kg-1 total ash. Mangrove

seeds are poor in trace elements such as copper, zinc and manganese (Faya

et al. 1992).

In the Middle East by the 13th century, , mangroves were established

sources of food, fuel, medicine, and tanning leather ( Saenger, 1985).

Presently, mangrove leaves are being used as a feed ingredient for dairy

cows, sheep, and poultry with some advantages over their common

commercial feeds (Jara, 1985).. Human as well, eats processed mangrove

seeds ( as sweetened stuffing for pastry) or un-processed(salted seeds).

5

Jara, (1985). However, the available information on use of mangrove

products as animal feed stuffs are very limited and poorly documented.

The aim of the present study was to evaluate the effect of partial and

totally replacment of yellow corn meal with mangrove seed meal in a

practical style feed for bluespot mullet Valamugil seheli fingerlings on

growth and body composition.

Bluespot mullet Valamugil seheli were selected in this study as an

experimental animal as it is a herbivorous fish ( uses carbohydrates

efficiently ) and has a high market value(Harrison, and Senou 1997 ).. It

inhabits coastal waters but enters estuaries and rivers where it feeds on

microalgae, filamentous algae, diatoms, and detritus associated with sand

and mud (Harrison, and Senou 1997 ).

6

Materials and Methods

Collection of fish and acclimation

Valamugil seheli fingerlings (more than 1200 0.5g fish) were

collected from the wild. One hundred fingerlings were stocked in each of

twelve tanks and gradually adapted on the new environment and artificial

feed for two weeks.

Mangrove seeds preparations:

Twenty kilograms of mangrove seeds were collected from trees in

the Annak site on the Arabian Gulf during the month of October. Seeds

were then air dried for four days followed by three days in a convection

oven at 60 oC. The dried seeds were then ground to powder form in a

commercial blender and analyzed for proximate composition, minerals,

carbohydrates (Table 1) and amino acids) (Table 2), and kept in a freezer

at -8oC until mixed into the test rations.

Experimental feed preparation:

Four approximately isoenergetic isonitrogenous rations with 0 g kg-1

, 150 g kg-1 , 300 g kg-1 and 510 g kg-1 mangrove seed powder (on dry

7

weight basis) were formulated (Table 3). The test diets contained the

minimum requirement of all essential nutrients to satisfy the needs of

marine warm water fishes as recommended by NRC (1993).The diets were

prepared as follows: all feed ingredients were ground in a commercial

blender and mixed in a kitchen mixer. Vitamin and mineral mixes were

gradually added with continuous mixing. Distilled water (60oC) was

slowly added while mixing until the mix began to clump. The diet was

then passed through a kitchen meat grinder and dried for 24 h at 60oC in a

convection oven. The dried diet was chopped into pellets in a blender and

then passed through laboratory test sieves (mesh 2 and 0.88 mm) to ensure

homogenous particle size of sinking pellets and stored at -8oC until used.

The amount of dust (under size material) as a result of the pelleting

process was recorded for each test feed as an indicator of friable or robust

pellets.

Feeding trial

Each feed was fed on a dry weight basis to satiation to triplicate

tanks of fish three times a day. Samples of twenty fish from each tank were

weighed every seven days and the trial continued for a period of ten weeks.

At the end of the experiment all the fish from each tank were separately

8

killed, weighed, ground in a commercial blender and stored at -8oC for

subsequent body composition analysis.

System description:

The system was a totally closed recirculating marine water system

consisting of twelve aerated cubic fiberglass tanks (1.5m W. x 1.5m L. x

1m H) with associated settling tanks, biofilteration and UV terilisation.

Seawater was obtained from the Arabian Gulf Half Moon Area,

disinfected using bleach (7 ml L-1) and salinity adjusted from 65 to 45 g L-1

using fresh water source. Water temperature was kept at 24± 2 oC.

Analyses

Mangrove seed and yellow corn analyses were performed according

to the methods described in AACC (1995). Total sugars and amino acids

analyses were carried out using HPLC (Model 1993, Shimadzu, Japan

according to manufacturer's methods. Minerals analyses were performed

according manufacturers methods using AAS – 680 systems for element

analysis. (Shimadzu, Japan).

Each diet and fish sample was analyzed for moisture, crude protein,

crude fat (ether extract), crude fibre (for feed samples only) and total ash

content in triplicate (AOAC (1980).

9

Water quality from all tanks were analyzed once every week for O2

using PSI DO meter, carbon dioxide using a CO2 meter, ammonia and

nitrite using the methods described by Boyed, (1978).

All data were analyzed using SAS ANOVA procedure (Statistical

Analysis 1995). One-way analysis of variance (ANOVA) and Duncan’s

multiple range tests were used to compare treatment means (Snedecor and

Cochran 1981). Statements of significant differences are based on P < 0.05.

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Results

Overall, no disease symptoms occurred throughout the experimental

period. Mortalities were small and not related to any treatment.(Table 4).

Water quality parameters (O2, CO2, pH, water temperature, and total

ammonia nitrogen) changed over the course of the experiment O2 decreased

from 7.9 to 6.2 mg L-1, CO2 increased from 3.55 to 6.25 mg L-1, pH was

slightly decreased from 7.7 to 7.23, water temperature gradually increased

from 24.1 to 29.2 0C and Total ammonia ranged from 0.01 to 0.72 mg L-1.

There were no significant differences between treatments as the system is

inter-connected.

Table 1 shows the proximate composition, minerals, starch and total

sugar content of both mangrove seeds and yellow corn. Mangrove seeds

contain slightly higher crude protein, higher crude fibre (double), much

higher total ash (four times), much higher total sugars (seven times),

slightly lower carbohydrates (nitrogen free extract), lower starch (two

third) and much lower (one forth) crude fat than those in yellow corn.

Additionally, mangrove seeds contain much higher levels of sodium,

potassium, copper, iron, and zinc.

Table 2 shows the amino acid content of mangrove seeds and yellow

corn. It was found that mangrove seeds contain lower levels of threonine,

11

glutamic acid, Leucine, lysine, and arginine than corn. Mangrove seed

composition indicates that it is mainly a carbohydrate source.

The proximate composition of the experimental diets (Table 3)

showed little variation in the nutrient levels of the various diets and agreed

with estimated values.

Good binding properties were noted with increasing levels of

mangrove seed in the experimental diets. The level of fines during the

pelleting process deceased (450 g kg-1, 310 g kg-1, 230 g kg-1 and 150 g

kg-1) for diets containing 0 g kg-1, 150 g kg-1, 300 g kg-1 and 510 g kg-1

mangrove seed respectively, with a very high correlation (r2 = 0.94, P

<0.05).

Growth and feed efficiency parameters are shown in Table 3. There

was no significant difference among the initial weights of fish (P > 0.05).

However, final weights and weight gain for all groups were significantly

different (P < 0.05) ranking the diets 300 > 150 > 0 > 500 g kg-1 mangrove

seed. Other growth and feed utilization parameters (specific growth rate

(SGR), feed conversion ratio (FCR), and percentage protein deposited

followed the same pattern as weight gain.

Fish fed the diet containing 510 g kg-1 mangrove seed performed

poorly (P< 0.05) relative to all other experimental feeds in terms of growth

and feed efficiency.

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Proximate carcass composition is shown in Table 5. Body

composition was affected (P < 0.05) by replacing dietary corn with

mangrove seed. As the level of the mangrove seed incorporation increased

in the test diets, carcass moisture, ash, and protein were increased, whilst

body fat gradually deceased.

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DISCUSSION

The closed recirculating culture system used in the expriment was

capable of maintaining suitable water quality parameters for experimental

fish (Wheaton et al. 1994)

Dietary carbohydrate sources (maize, sorghum, wheat, rice and

barley ) are important sources of dietary energy for herbivorous fishes such

as carp and tilapia as they can utilize high levels of digestible carbohydrate

(Anderson et al 1984, Teshima et al. 1987 ; El-Sayed and Garling

1988).Valamugil seheli, a predominantly herbivorous fish, are expected to

be capable of utilizing the carbohydrate and fibre from corn and mangrove

seeds in the experimental diets (Harrison, and Senou, 1997 ).

It was indicated that the incorporation of Mangrove in Valamugil

seheli feed as a replacement for corn at 300 g kg-1 mangrove level produced

superior effect on growth rate, feed conversion, specific growth rate,

protein efficiency ratio and percentage protein deposition as compared to

all other mixing levels.

That could be explained as followed:

1- Corn and mangrove contain different levels carbohydrate forms

(starches, sugars, ) (Table 1). The rate of mixing corn and mangrove

changes levels of carbohydrates and may affect the rate of

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carbohydrate degredation( glucose release ) to be faster or slower in

fish gut. Glucose is known to inhibit the transport of amino acids at

absorption site in fish gut membranes when it is released too fast

(Hokazono 1979 ). It is possible that the inhibition of amino acids

transport account for inferior protien retention of fish(tilapia )

Anderson et al. (1984). Therefore, when the level of carbohydrates

mix is optimal, the rate of glucose relrease may not affect affect

amino acids transport, protein retention and produces superior

growth rat. That was supported by higher percentage of protein

deposited and protein efficiency ratio and growth rate at the optimum

corn : mangrove mixing ratio 30:15 (2:1) than all other ratios.

(Table) 4. Similar results were indicated when different dietary

carbohydrates ingreadient mixes were fed to chicken (Kamel et al.

1981; Najib and Al-Beshr 1986) and fish ( Belal and Al-Jasser

(1997); Belal (1999).

2- Additionally, mangrove seed and corn have different starches in

terms of their physical characteristics such as size and /or shape.

Mangrove starch is gelatinized at different temperature to that from

maize, wheat, or rice (62-72 oC). This increases mangrove seed

viscosity and improves pellet quality (Lineback 1984). As a result,

mangrove has a much more viscous effect than maize on fish pellet

15

quality. In other words, the pellets containing mangrove do not need

as much, if any, binder as with many other grains (maize, wheat etc.).

Gabandan (1986) demonstrated that using high level of binders (high

viscosity) in fish feed pellets may reduce their digestibility by as

much as 150 g kg-1. Additionally, starch granules in corn have more

surface area, exposed to more to enzymatic hydrolysis (Lineback

1984), because they are smaller those that of mangrove seeds.

As a result, the rate of starch degradation (glucose release) would be

affected by corn mangrove mixing level. Consequently, growth rate

of fish will be affected as, it is explained above. As a result, the

growth rate of fish a fed diet containing an optimum mangrove seed

to corn ratio would be higher than any other ratios.

When Valamugil seheli were fed a diet-containing mangrove seeds

alone, they grew more poorly than fish fed a diet containing corn alone or

corn-mangrove mixed as a carbohydrate source. This is explained as

follows: Stronger pellets that produced by higher level of mangrove seeds

(510 g kg-1), which lower the diet’s digestibility.

Body composition analysis indicated that as mangrove seeds

incorporation in the feed were increased, body moisture and protein

deposition were increased and body fat was decreased. That could probably,

be explained by improving the protein sparing effect of the dietary

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carbohydrates during protein synthesis as indicated from the percentages of

protein deposition. Higher moisture content causes a reduction of the

percentage of body fat. Additionally, higher levels of mangrove seeds may

have reduced digestibility by producing strong pellets. This resulted in an

increase of body moisture and a reduction in fat deposition and growth rate.

In conclusion, mixing mangrove seed with corn in Valamugil seheli

feed would not affect the fish growth parameters. However an optimum

mixing of a mangrove: corn ratio 30:15 (2:1) in Valamugil seheli diet,

would result in superior growth parameters under the experimental.

Cheaper and more efficient fish diet could be produced using

mangrove seed. This because mangrove plant is highly productive and sea

water irrigated as compare to costly fresh water ones for many countries

around the world.

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El-Sayed, A. M., and Garling, D.L. (1988). Carbohydrate to lipid ratio

in diets fed to Tilapia zilli fingerlings. Aquaculture (73), 157-163.

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Hamilton, L.S. and Sendaker, S.C., (1984) Handbook for Mangrove Area

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Center, Environment and Policy Institute, Hawaii, USA, p 2-37.

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Harrison, I.J. and Senou, H. (1997) Mugilidae. Mullets. In K.E. Carpenter

and V. Niem (eds.) FAO Identification Guide for Fishery Purposes. The

Western Central Pacific, referance no. 9812, p 41-42.

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mossambicus Aquaculture 27, 97-109.

Jara, R.S. (1985). Traditional Uses of Mangroves in the Philippines. In

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Dartnall.Eds.). Proceeding of the Research for Development Seminar.

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21.

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Eds.). Proceeding of the Research for Development Seminar.

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rate, fish size and dietary protein and cellulose levels on growth of

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21

methods. In Development of Aquaculture and Fisheries Science, V. 27,

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(Halver, J. E.ed.) pp 142, Academic Press, INC. San Diego, California,

USA.

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Table 1

Analysis of mangrove seeds and yellow corn seeds (dry weight).

_________________________________________________________

Type of analysis Mangrove seeds Corn1

_________________________________________________________ Moisture (g kg-1) 33.9 139 Crude protein (g kg-1) 108 93.7 Crude fat (g kg-1) 84 41.4 Crude fiber (g kg-1) 58 24.2 NFE (g kg-1) 773.6 827.5 Starch (g kg-1) 422 6932

Total sugars (g kg-1) 140.5 20.9 Total ash (g kg-1) 52 13.2

• Calcium (mg kg-1) 2 779 800 • Phosphorus (mg kg-1) 2 0.21 0.26 • Potassium (mg kg-1) 2 1.61 0.25 • Sodium (mg kg-1) 2 0.88 0.03 • Magnesium (mg kg-1) 2 976 747 • Manganese (mg kg-1) 2 5.6 4.40 • Copper (mg kg-1) 2 9.4 2.14 • Iron (mg kg-1) 2 84,99 25.0 • Zinc (mg kg-1) 2 231 21

_______________________________________________________ 1 Yellow corn grad 2 from The USA 2 Performed using Atomic Absorption Spectrophotometer according to AACC (1995)

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Table 2 Amino acids content of measured mangrove seeds and yellow corn are

indicated (NRC 1993 ) as g kg-1 protien.

Amino acids Mangrove seeds Yellow corn

Arginine

Histidine

Isoleucine

Leucine

Lysine

Methionine

Phenylalanine

Tyrosine

Threonine

Valine

Alanine

Asparatic acid

Glutamic acid

Glycine

Serine

0.81

0.23

0.27

0.56

0.17

0.19

0.59

0.42

0.38

0.59

0.45

0.81

0.92

0.54

0.41

0.41

0.24

0.27

0.98

2.9

0.20

0.42

0.28

0.20

0.42

0.62

0.58

1.56

0.34

0.31

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Table 3 Proximate Composition and proximate analysis of the test diets are

indicated based on dry weight.

a. Control b. Peruvian fish meal, 65% crude protein, 12% crude fat, from

Nutris Co., S.A., 3 Rue, Rosenwald, 75015 Paris, France. c. As reported by Jackson, Capper & Matty (1982) d. Nitrogen-Free Extract, determined by difference. e. Gross energy, calculated based on 23.67, 17.17 and 39.79 kJ g-1

protein, carbohydrate and lipid, respectively.

Feed ingredients Mangrove seeds content in the test diets (Dry weight) 0 g kg-1 150 g kg-1 300 g kg-1 510 g kg-1

a

Fish meal b 410 410 410 410 Yellow corn 510 360 210 000 Mangrove seeds 000 150 300 510 Corn oil 00 0 5 10 15 α Cellulose 20 15 10 5 Vitamins Premix c 20 20 20 20 Minerals premix c 20 20 20 20 Di-calcium phosphate 20 20 20 20 Proximate analysis g kg-1

• Crude protein 316 319.5 322 316 • Crude fat 69.5 70.1 68.7 67.8 • Crude fiber 38.4 36.2 34.2 37.4 • Total ash 126.4 127.7 129.8 136.5• NFE d 449.7 446.5 445.6 442.3• Energy e(kJ g-1) 179.7 180.2 180.1 177.7

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Table 4

Growth, feed utilization, feed conversion and the percentages of survival of Valamugil seheli fingerlings fed the experimental diets. Values expressed as a mean of three replicate groups' ± SE (superscript) g kg-1 of fish wet weight. (P < 0.05)

Mangrove content in

diet (g kg-1 dry wt)

Mean initial wt. (g fish-1)

W1

Mean final wt. (g fish-1)

W2

Weight gain (g fish-1) W2-W1

Feed intake

(g fish-1)

FCR a

SGR b

PER c

Percentage protein

deposited d

Survival %

0

0.49±0.2

13.92± o.62

13.43±0.47

28.34

2.12

4.78±0.08

1.51±0.11

20.00

96.8

150 0.5±0.1 15.3± 0.46 14.8±0.36 26.64 1.98 4.89±0.11 1.73±0.06 23.00 98.3

300 0.48±0.2 18.0± 1.05 17.52±0.76 30.3 1.73 5.18±0.18 1.80±0.09 27.97 97.2

510 0.52±0.2 11.38±0.95 10.86±0.58 26.61 2.45 4.40±0.28 1.29±0. 14 23.66 99.5

a. FCR, Feed conversion ratio (feed intake/average weight gain per fish for the ten week period) b. SGR, specific growth rate = [(In W2-In W1)/time in days) x 100] c. PER, protein efficiency ratio = average weight gain (g)/average weight of protein fed (g). d. Percentage protein deposited = [ ( final body protein – initial body protein ) x 100]/ totat proein fed. Wilson, R.

P. (1988).

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Table 5 Whole body composition1 of mullet Valamugil seheli fingerlings fed the

experimental diets.

1Expressed as a mean of three replicate groups' ± SE (superscript) g kg-1 of fish wet weight. (P < 0.05)

Type of analysis

Initial body composition

g kg-1

Mangrove seeds content in diet 0 g kg-1 150 g kg-1 300 g kg-1 510 g kg-1

Moisture

708.3

674.4±5.7

667.2± 4.2

682.1± 3.7

701.6± 5.6

Crude protein

159.1

135.7± 5.7

146.6± 8.9

160.9± 4.8

183.2± 6.4

Crude fat 69.8 114.9± 2.9 99.3± 1.7 95.0± 2.0 84.3± 2.4

Total ash 57.14 47.8± 0.7 47.4± 0.5 53.4± 0.9 63.39± 0.9