biochemical composition of zooplankton community.pdf

8/11/2019 Biochemical composition of zooplankton community.pdf

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Biochemical composition of zooplankton community grown in

freshwater earthen ponds: Nutritional implication in

nursery rearing of fish larvae and early juveniles

Gopa Mitra a ,, P.K. Mukhopadhyay a, S. Ayyappan b

a Central Institute of Freshwater Aquaculture, Kausalyaganga, Bhubaneswar-751 002, Orissa, Indiab Krishi Anusandhan Bhawan-II, ICAR, Pusa, New Delhi-110 012, India

Received 8 November 2006; received in revised form 7 August 2007; accepted 8 August 2007

Abstract

This study was conducted to obtain a database describing the nutritional value of freshwater mixed zooplankton that are widely

used for larval and grow-out rearing of freshwater fish. The macro and micronutrient composition of mixed zooplankton samples

collected from 6 fertilized earthen ponds were analysed for protein, lipid, carbohydrate, ash and these ranged from 73 79%, 10.79

14.55%, 34.79% and 3.2010.07%, respectively on a dry matter (DM) basis. Amino acid profile showed the presence of all the

ten essential amino acids with low level of methionine. The content of saturated fatty acids (SAFA), mono unsaturated fatty acids

(MUFA) and polyunsaturated fatty acids (PUFA) ranged from 6481%, 715% and 620% of total fatty acids, respectively. The

predominant fatty acids were SAFA (16:0), MUFA (18:1n-9), PUFAviz.linoleic acid (LA 18:2 n-6) and linolenic acid (LNA 18:3n-3). Among the vitamins, ascorbic acid (1540.01 g/gDM) was less than the requirement of fish especially for larvae, vitamin-A

(13.6163.95 g/g DM) and vitamin-E (218348g/g DM) were more than the requirement of fish. Mineral and trace element

content showed the presence of P, Ca, Fe, Cu, Zn and Mn. Seasonal variation of all biochemical components was evaluated in the

study. Vitamin E had strong co-relation (r1=0.72; r2=0.88; r3=0.83; r4=0.86; r5=0.36 and r6=0.88) with seasonal variation in

lipid content of zooplankton of different ponds and varied inversely with that of rising temperature. Enzyme content from the

mixed zooplankton of different ponds showed availability of protease (6.217.92 g leucine/ mg protein/h), lipase (25.8239.1 g

-naphthol/mg protein/h) and amylase (100226.1 g maltose/mg protein/h), which could be used as an exogenous source of

digestive enzymes for fish and shellfish during ontogenesis. Absence of l-gulonolactone -oxidase activity confirmed the

incapability of these zooplankton to synthesize ascorbic acid (AA) de novo. The average dry weight in zooplankton in different

ponds was 1.24.2 mg/l and different species present in these ponds were Moina (Moina dubia), Daphnia (Daphnia lumholtzi,

Daphnia carinata); Cyclops (Mesocyclops hyalimus, Mesocyclops leuckarti); Diaptomus (Heliodiaptomus viddus, Neodiaptomus

handeli); Rotifer (Brachionus). These results indicate that the biochemical composition and subsequently the nutritional value ofthese planktonic organisms are not only genetically determined but also influenced by its maturity stage and type of ingested food.

These data may be helpful for reference purpose and for formulated feed preparation accessing the nutritional potentiality of these

freshwater zooplankton in the nursery rearing of freshwater fish larvae and early juveniles.

2007 Elsevier B.V. All rights reserved.

Keywords: Mixed zooplankton; Protein; Lipid; Vitamins; Minerals; Amino acids; Fatty acids and enzymes

Available online at www.sciencedirect.com

Aquaculture 272 (2007) 346 360www.elsevier.com/locate/aqua-online

Corresponding author. Tel.: +91 674 2465446; fax: +91 674 2465407.

E-mail address:[email protected](G. Mitra).

0044-8486/$ - see front matter 2007 Elsevier B.V. All rights reserved.doi:10.1016/j.aquaculture.2007.08.026
mailto:[email protected]://dx.doi.org/10.1016/j.aquaculture.2007.08.026http://dx.doi.org/10.1016/j.aquaculture.2007.08.026mailto:[email protected]


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1. Introduction

Zooplankton occupy a central position between the

autotrophs and other heterotrophs maintaining an im-

portant link in the sustainability of the food chain form-

ing one of the most important components of freshwateraquaculture species (Chakrabarti and Sharma, 1998).

Larvae of all cultivable fish species require live plank-

tonic organisms as first food (Garcia-Ortega et al., 1998).

In semi-intensive or intensive culture conditions, aqua-

culture species derive a substantial part of their dietary

nutrient needs from naturally available zooplankton as

they are a valuable source of protein, amino acids, lipid,

fatty acids, vitamins and enzymes (Millamena et al.,

1990; Munilla-Moran et al., 1990; Pillay, 1990; Evjemo

et al., 2001). Thus the relative contributions of zoo-

plankton in the nutrition of freshwater fish larvae haveimmense significance (Jana and Chakrabarti, 1993).

Despite the effort that has been put in the development of

formulated starter feed (Verreth et al., 1987) for larval

fish, live food still remains a better option in terms of

survival and growth compared to formulated diet alone.

Live food seems to provide a good source of exogenous

enzymes, and also helps in chemoreception and visual

stimuli (Kolkovski et al., 1995). However, the nutritional

quality of zooplankton varies considerably and thus

plays a major role in producing quality larvae and juve-

niles (van der Meeren et al., 2001) as well as they would

aid in determining the suitability of the organisms in fishlarvae culture (Kibria et al., 1999).

Although in semi-intensive fish culture the cultured

species draws a significant part of nourishment from zoo-

plankton grown in ponds, however quantitation of its

nutritional contribution to fish growth is limited. However

the dependence on live food as starter feed rather than on

formulated feed in fish larvae makes it pertinent to evaluate

the nutritional composition of live food in aquaculture

(Srivastava et al., 2006). Due to escalation in the cost of

Artemia cysts, generally used during larval rearing, use

of pond grown zooplankton is justifiably gaining moreimportance in the hatcheries in different regions (Evjemo

et al., 2003; Velu and Munuswamy, 2003). However,

colonization of planktonic organism can be found in si-

milar water bodies of different countries (Welch, 1952).

Studies were thus conducted to determine the proximate

composition, content of amino acids, fatty acids, vitamins

(A, C and E), minerals (P, Ca), trace elements (Fe, Cu, Zn,

Mn) and certain metabolic enzymes of mixed zooplankton

from different freshwater fish ponds. Also their nutritional

contribution are evaluated with the aim to consider the

nutritive potentiality of this zooplankton for nursery

rearing of carp larvae and early juveniles.

2. Materials and methods

2.1. Pond preparation and collection of samples

Zooplankton samples were collected twice every month

from 6 culture ponds (area 0.04 ha, mean depth 1.5 m,) of CIFA

farm at Kausalyaganga, Orissa, India (Lat 20 20N; Long 85

49 E) for one year. The pre-sampling pond preparation was

carried out for eradication of predatory and undesirable fish

according toJena et al. (1998). The ponds were dried initially

and exposed to sunlight for eradication of predatory and unde-

sirable fish prior to stocking and filling with canal water filtered

through nylon net (40 mesh) for preventing the entry of pre-

datory species. Pond fertilization was carried out according to

Jena et al. (1998)with application of raw cow dung (0.39% N

and 8.0% C) and super phosphate at 3 tonnes/ha and 7 kg/ha,

respectively, as a basal dose with alternating applications every

fortnight at 1 tonnes/ha/month and 20 kg/ha/month, respec-

tively. Water levels in the ponds were maintained at 1.4to 1.5mthroughout the period compensating for seepage and evapora-

tion losses.

The samples were collected between 0800 and 0900 h by

filtering 200 l of water using plankton net made of bolting silk

(0.06 mm mesh size) from four sites of each pond with a plastic

container (5 l) from the subsurface (at 0.50.6 m depth) water

with least disturbance. By passing the water of the plastic

container through a net (0.064 mm), the plankton density was

increased (any plankton stuck to the side of the net was washed

down by gently splashing water on the outside of the net) and

transferred to a 10 l plastic container (having 9 l water) through

a 1050 m mesh net (to exclude large predatory insects like

notonectids). Then it was thoroughly mixed to get a represen-

tative of a composite sample of each pond. The samples were

then brought to the laboratory for analysis of nutritive compo-

sition. Prior to analysis of nutritive composition, the plankton

was suspended at a density of 150200 numbers/ml in tap

water in 20 l well aerated glass jars. Dead plankton and organic

debris were removed by siphoning with 2 mm diameter pipe.

Before preparation for biochemical analysis, the samples were

checked for zooplankton viability and no organic debris were

present in the samples. After 24 h the zooplankton samples

were harvested by sieving (through mesh #25 bolting silk) the

water from each glass jar and rinsed in distilled water. After

harvesting some portion of the zooplankton samples were driedat 60 C to constant weight to determine the dry matter content

from which protein, ash and amino acids were analysed. Sub-

samples of harvested zooplankton were packed and immedi-

ately frozen in liquid nitrogen and stored at 80 C until

analysed for lipid, fatty acids, enzymes and vitamins. From all

samples a known volume was immediately fixed in 4% forma-

lin for quantitative estimation of plankton species.

2.2. Nutrient composition

Dry matter (DM), crude protein (CP) and ash content were

determined according to the standard methods (AOAC, 1998).DM content was determined after drying in an oven at 60 C to

347G. Mitra et al. / Aquaculture 272 (2007) 346360


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constant weight and crude protein (CP) content by the Kjeldahl

method and calculated as nitrogen content multiplied by 6.25.

Lipid was analysed using the method of Bligh and Dyer

(1959). Isolated zooplankton was homogenised with 15 ml

chloroform:methanol (2:1 v/v) containing 0.05% butylated

hydroxy toluene (BHT) as an antioxidant in a 20 ml glass

homogenizer. Lipid extracted was re-dissolved in chloroform/

methanol (2:1, v/v containing 0.05% BHT) at a concentration

of 10 mg/ml and stored under nitrogen atmosphere at20 C

until fatty acid analysis.

Ash content was determined by incinerating dried samples

in muffle furnace at 550 C for 3 h and then preserved for

mineral estimation. Organic matter (OM) was calculated by

subtracting the total ash value from DM. Total carbohydrates

(TCHO) was determined by subtracting CP and lipid values

from OM.

2.3. Amino acid

For amino acid analysis defatted plankton samples were

dried under vacuum in a HPLC work station (Model: 2690) for

30 min (Waters, USA). After purging with nitrogen the sam-

ples were hydrolysed with 6N HCl at 110 C for 20 h under

vacuum. Hydrolysed samples were dried and redried with a

mixture of ethanol:water:triethylamine (TEA) (2:2:1) at

(6586.13/760) N/m2. Phenyl isothiocyanate (PITC) deriva-

tives were prepared by adding PITC reagent (ethanolTEA

WaterPITC:: 7:1:1:1) and mixed well and incubated at room

temperature for 30 min, the samples were then allowed to dry

under vacuum. Diluent was added in each sample, which was

then processed for filtration (45 filter paper) and analysed.

Operating condition was: column temperature: 38 C: column:

pico-tag, absorbance 254 nm, pump pressure 1000 psi.

The chemical score (CS) was calculated based on the

limiting essential amino acid in zooplankton multiplied by 100,

where limiting essential amino acid is that which has the

following lowest ratio of essential aminoacid in plankton:

essential amino acid in fish tissue protein (Hepher, 1988).As in

other animals, arginine, histidine, isoleucine, leucine, lysine,

methionine, phenylalanine, threonine, tryptophan and valine

were considered the essential amino acids (Tacon and Cowey,

1985). The non-essential amino acids cystine and tyrosine can

only be synthesized by the fish from methionine and phenyla-

lanine, respectively (Tacon and Cowey, 1985). Therefore, themethionine and phenylalanine requirement of the fish will

partially depend on the cystine and tyrosine content of the diet.

For the calculation of the CS, the values of methionine and

cystine were summed and taken as one essential amino acid in

fish. The same was carried out with phenylalanine and tyrosine.

Tryptophan was not considered in this calculation because it

was not determined.

2.4. Fatty acid

Fatty acid methyl ester (FAME) derivatives of the fatty

acids were prepared by adding 10 ml 20% borontrifluoride(BF3) in methanol to 100 mg lipids and heating for 30 min at

100 C. After cooling, water (same volume as the BF3) and

hexane (half the volume of BF3) were added, and centrifuged

at 3000 rpm for 5 min. The content of the centrifugation tube

was then transferred into a small separating funnel and allowed

to separate for 510 min. The lower phase was discarded and

the upper phase was recovered in new flasks and FAME in

hexane were dried with sodium sulfate and the sodium sulfate

was removed by filtering through a Pasteur pipette containing

glass wool. The operating conditions of the gas chromatograph

(PYE UNICAM, GC 104) were: flame ionization detector

(FID), stainless steel column packed with 10% diethylene gly-

col succinate polyester (DEGS), column temperature: 195 C

(maintained for 10 min isothermal), injection port temperature:

210 C, detector temperature 210 C, carrier gas (N2) with 35

40 ml/min flow rate, recorder chart speed 640 mm/h. The

methyl ester peaks were identified by co-chromatography with

standard fatty acid methyl ester (Sigma Chemical Co, USA)

mixtures by comparing their retention time with the retention

time of known fatty acids and quantified by a Spectraphysic SP4270 integrator.

2.5. Analysis of vitamins

Ascorbic acid content in sub samples was determined

according toRoe and Kuether (1943)modified byDabrowski

and Hinterleitner (1989). The plankton were homogenised in

5% trichloroacetic acid (TCA) solution containing 250 mM

(HClO4) and 0.08% ethylenediamine tetra acetic acid (EDTA)

using an motor driven glass Teflon homogeniser in ice and

centrifuged at 29,000 g for 30 min at 4 C. Total ascorbic acid

content in tissue homogenate was measured by 2,4 dinitro

phenylhydrazine (DNPH) method byRoe and Kuether (1943)

modified byDabrowski and Hinterleitner (1989)in which 2, 4

dinitrophenyl hydrazine derivative of ascorbate was measured

spectrophotometrically at 524 nm. Modification of original

method included incubation with dichloroindophenol (DCIP)

shortened to 20 min, incubation temperature was 30 C and

additional blank per sample was included to account for in-

terfering substances according toDabrowski and Hinterleitner

(1989). For dry weight measurement sub samples of plankton

collected from same batch were oven dried at 60 C for 24 h till

constant weight.Vitamin A of sub zooplankton samples was analysed

according to the method ofSoni et al. (1997). Known amountof zooplankton was homogenised with chloroform (CHCl3).

The filtrate was added with benzene:toluene (2:1v/v) fol-

lowed by acid mixture containing H2SO4: glacial acetic acid

(1: 4 v/v) which gave blue colour. The blue colour initially

produced immediately changed to pink colour and the solu-

tion remained covered with benzene or toluene, which stabi-

lizes the colour by probably preventing the oxidation of

chromophore. The max was measured at 525 nm against a

reagent blank.Vitamin E was extracted following the method Quaife and

Dju as described byOser (1960)and estimated by the method

of Emmerie-Eangel Reaction (ferric chloride-dipyridyl meth-od). Extracted homogenate was mixed with ferric chloride

348 G. Mitra et al. / Aquaculture 272 (2007) 346360


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reagent (0.1 g FeCl3dissolved in 50 ml absolute ethanol) and

0.5% dipyridyl in absolute ethanol. The mixture was mixed by

swirling. The mixture was again mixed with absolute ethanol

and shaken vigorously. The max measured at 520 nm against

a reagent blank.

2.6. Mineral analysis

The ash was moistened with a small amount of glass-

distilled water and 5 ml of 6 N hydrochloric acid (AR grade)

was added. Calcium was measured by flame photometry, P

content by spectrophotometry following Fiske and Subbarow

method (Fiske and Subbarow, 1925) and trace elements (Zn,

Cu, Fe and Mn) by atomic absorption spectrophotometry

(AAS).

2.7. Enzyme assays

The total digestive enzymes: protease, lipase and amylasewere measured in the pooled zooplankton homogenates. For

homogenate preparation 100 mg sample was ground in a potter

blender with 0.89% NaCl solution. After homogenisation the

samples were sonified for 30 s and centrifuged at 12,000 g for

10 min at 4 C. During preparation the homogenates were

continuously kept on ice.

Total protease activity was measured in a medium contain-

ing 0.05 M trisHCI buffer (pH 8.0) using bovine serum

albumin (0.5 mg/ml) as substrate. The assay mixture consisted

of 200 l BSA solution, 100 l enzyme solution and 200 l

buffer was incubated at 37 C for 30 min, the reaction was

stopped with addition of cold trichloroacetic acid (10%). The

enzymatically liberated amino acids were assayed according to

the method ofMarks and Lajtha (1963). The results are ex-

pressed as g leucine liberated per mg protein in sample per

hour. Protein content of the supernatant solution was deter-

mined by the method of Lowry et al. (1951), using bovine

serum albumin (BSA) as standard.

The assay of amylase activity was based on method of

Bernfeld (1955). The activity is expressed as mg maltose

liberated from starch/g protein/h.

Activity of lipase was measured using the method of

Seligman and Nachlas (1963) using the substrate, 0.2% -

napthyl laurate in acetone: water (1:9 v/v). A unit activity is

defined as g -napthol liberated from -napthyl laurate in

60 min at 37 C. Specific activity is expressed as lipase activity

per mg protein (Lowry et al., 1951).Activity of l-gulonolactone -oxidase was measured using

the method as described by Mukhopadhyay et al. (1998).

Known quantity of plankton was homogenised in a glass ho-

mogeniser with 2 ml 50 mM phosphate buffer (pH 7.4) con-

taining 1 mM EDTA and 0.2% sodium deoxycholate. The

supernatant was assayed spectrophotometrically for ascorbic

acid by method described above. Protein was assayed in super-

natant by the method ofLowry et al. (1951).

Table 1

Composition of mixed zooplankton collected from different pondsPond 1 Pond 2 Pond 3 Pond 4 Pond 5 Pond 6

Proximate composition

Dry matter (DM) 11.81 2.10c 11.012.61c 10.093.18c 9.592.22b 7.410.49a 10.493.11c

Crude protein (% of DM) 79.91 1.6d 78.911.97c 78.851.65c 76.142.3b 76.480.98b 73.153.87a

Crude lipid (% of DM) 12.07 2.98b 13.701.89c 12.212.4b 10.793.6a 14.550.97d 13.91.87c

Total carbohydrate(% of DM) 4.792.56a 4.111.99a 4.061.79a 3.002.1a 3.092.6a 3.622.98a

Ash (% of DM) 3.22 1.33a 3.283.68a 4.883.93b 10.071.26e 5.881.98c 9.330.98d

Enzymes

Amylase (g maltose/mg protein/h) 241.22 7.10e 222.635.61d 109.394.42a 105.974.33a 153.923.12b 185.444.81c

Protease (g leucine/mg protein/h) 6.710.21ab 6.540.21a 7.570.25c 6.820.12b 6.770.19b 7.610.29c

Lipase (g naphthol/mg protein/h) 34.00 0.37a 35.380.46ab 37.680.65c 36.450.57b 35.650.33ab 35.162.98a

Vitamins

Vitamin A (g/gm DM) 21.84 0.976b 52.041.15d 62.311.50f 15.591.39a 53.931.46e 47.371.05c

Vitamin E (g/gm DM) 230.63 18.4a 267.2215.06b 258.6611.26b 228.937.58a 333.4710.4d 319.3911. 06c

Vitamin C (g/gm DM) 15.16 0.16a 15.410.34a 17. 210.88b 24.014.61c 39.070.64d 38.670.74d

Minerals

Ca (mg/100 gm DM) 1.66 0.19b 1.361.16a 1.740.07b 2.360.43c 2.640.17d 2.770.24d

P (mg/100 gm DM) 0.86 0.20c 0.370.07b 0.940.09d 0.320.08ab 0.330.03ab 0.240.05a

Mn (mg/100 gm DM) 0.31 0.21d 0.020.0a 0.210.11c 0.080.02b 0.390.03e 0.340.24d

Zn (mg/100 gm DM) 1.01 0.05b 1.020.07b 0.060.03a 2.580.17d 1.310.21c 2.640.19d

Fe (mg/100 gm DM) 0.84 0.04d 2.200.32e 0.320.06b 0.130.03a 0.530.03c 0.160.01a

Cu (mg/100 gm DM) 0.08 0.01b 0.070.07b 0.160.17c 0.080.14b 0.070.02b 0.050.02a

Values are given as meanStandard deviation (SD) (n =12). Mean values with different superscripts in a row are significantly (Pb

0.05) differentfrom each other.



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2.8. Collection of water sample and qualitative analysis of

plankton

Regular fortnightly sampling of water from the above aqua-

tic bodies was done in the morning (08:00 h) and relevant

physico-chemical parameters were analyzed. The phosphate-

phosphorus (PO4P) content of the water was estimated by the

stannous chloride method (Clesceri and Greenbreg, 1989), pH

by Elico digital pH meter L 1120, free CO2(titrimetric method

using N/44 Na2CO3), alkalinity (titrimetric method using

0.02N H2SO4; Clesceri and Greenbreg, 1989), Nitrite-N (NO2N) and Nitrate nitrogen (NO3N) (spectrophotometric diazo-

tization method;Strickland and Parson, 1984), total ammonia

nitrogen (TAN) (phenolhypochlorite method), and O2by the

method described in Stirling (1985). Plankton content and

species identification was done as described inAPHA (1989).

Table 2

Amino acid content (% of total protein), chemical score of mixed zooplankton from different ponds and essential amino acid requirements of different

carps

Pond-1 Pond-2 Pond-3 Pond-4 Pond-5 Pond-6 Catla Rohu Mrigal Common carp

Arg 8.13 0.05f 7.440.07d 6.900.05b 8.010.11e 5.180.03a 6.980.03c 4.8 5.8 5.84 6.2

Lys 10.30 0.07a

12.550.04c

14.110.06e

13.700.03d

15.310.04f

12.440.03b

6.2 5.7 7.58 7.0His 1.97 0.03d 1.720.05a 1.880.04bc 2.060.23e 1.840.04b 1.950.03cd 2.5 2.3 3.02 2.9

Ile 3.74 0.06ab 3.920.04c 3.890.08c 3.720.03a 3.990.07d 3.780.03a 2.4 3.0 3.69 3.8

Leu 6.93 0.04a 7.070.07c 7.030.05b 7.020.04b 7.150.03d 7.480.03e 3.7 6.93 7.2

Val 4.50 0.11a 4.760.06c 4.790.05c 4.620.05b 5.130.04d 4.790.04c 3.6 4.32 4.5

Met 1.98 0.16c 1.960.06c 1.780.09b 2.060.05d 1.660.04a 1.690.05a 3.6 2.9 1.55 2.7

Phe 3.23 0.05a 3.530.08b 3.520.13b 3.220.04a 3.85.0.03c 3.910.04d 3.7 4.0 8.07 6.5

Thr 3.78 0.09b 4.040.11c 4.110.06d 4.170.04e 3.300.05a 4.210.05e 5.0 3.99 4.2

Tyr 5.54 0.09d 5.460.11c 5.620.05e 5.690.03f 3.920.06a 4.900.03b

Asp 7.32 0.05c 6.970.07b 7.480.04d 8.170.04e 6.730.04a 8.310.03f

Glu 12.57 0.05c 12.440.08b 12.140.05a 13.380.03e 12.770.04d 12.170.03a

Ser 3.29 0.04b 3.360.07c 3.460.09d 4.060.05e 2.860.04a 4.220.02f

Gly 5.83 0.03a 6.570.05f 6.410.13d 6.500.07e 6.090.04c 5.930.03b

Ala 7.18 0.03c 7.360.06d 7.420.07e 6.960.03b 7.810.05f 6.470.03a

TAA 86.29 89.15 90.54 93.34 87.59 89.23 TEAA 50.10 52.45 53.63 54.27 51.33 52.13

Chemical score (methionine)

Catla 55.00 54.44 49.44 57.22 46.11 46.9

Rohu 68.27 67.58 61.4 71.03 57.24 58.27

Common carp 73.33 72.59 65.92 76.62 61.48 65.00

TAA total amino acid, TEAA total essential amino acids.

Values are given as meanStandard deviation (SD). Means bearing different superscript in a row differ significantly (Pb0.05).

Table 3

Fatty acid composition of mixed zooplankton from different ponds

Fatty acids Pond-1 Pond-2 Pond-3 Pond-4 Pond-5 Pond-6

14: 0 7.28 1.59d Trace 2.02 0.87b 2.291.20b 0.230.08a 4.921.08c

16: 0 63.6 14.95a

66.4515.6a

64.1614.8a

70.2510.39a

81.23.34b

70.035.55a

16: 1 0.73 0.03c 0.100.0a 1.010.01d 0.200.01b 1.10.05e 2.10.04f

18: 0 2.12 0.7c 2.011.18c 2.340.89c 1.150.61b 0.10.0a 0.160.09a

18: 1 6.30 0.27a 12.1310.34a 13.829.85b 7.146.72a 8.32.54a 6.424.15a

18:2 n-2 13.44 6.89a 12.154.40a 10.564.53a 16.55.82b 10.12.19a 10.12.52a

18:3 n-3 6.34 3.03a 5.252.61a 4.602.17a

20:0 Trace Trace 2.510.73a 2.10.74a

20:1 4.122.95

Unidentified 0.180.02a 2.400.16b 2.00.03b

SAFA 73.02 14.8ab 68.4617.11a 68.5215.48a 76.212.08ab 81.33.38b 77.217.12ab

MUFA 7.03 6.7a 12.219.96a 14.759.97b 7.346.92a 8.182.66a 12.576.25a

PUFA 19.78 9.47b 17.46.94b 15.166.28ab 16.55.00b 10.12.17a 10.12.51a

n-3/n-6 0.47 0.43 0.44

SAFA

Saturated Fatty Acid, MUFA

Monounsaturated fatty acid, PUFA

Polyunsaturated fatty acid. Values are given as mean Standarddeviation (SD) (n =12). Mean values with different superscripts in a row are significantly (Pb0.05) different from each other.



6/15

2.9. Statistical analysis of data

Data are presented as meansstandard deviation of means.

To correlate vitamin E content in the zooplankton with lipid

content correlation coefficient (R value) analysis between these

two parameters was performed using microsoft excel software.

One-way analysis of variance (ANOVA) with multiple means

tests DMRT (Duncan's Multiple Range test,Duncan, 1955) in

respect of ponds was applied over all the parameters to inves-

tigate the difference existing between the ponds.

3. Results

The protein, lipid, carbohydrate and ash compositions of

zooplankton are given inTable 1in relative % of dry matter

(DM) values. The relative concentration of zooplankton DM

was on average 9.61% of wet weight through the whole sam-

pling period, varying between 7.4111.81%. Protein content

was relatively higher in the zooplankton of pond 1, 2 and 3

than other ponds and varied from 7379% of DM. The total

lipid content was between 10.7914.55% of DM. The magni-

tude of seasonal variation in lipid content was relatively high

in zooplankton of different ponds and it was inversely related

to environmental temperature. Carbohydrate content varied

between 34.79% and ash content 3.2210.07% of DM, res-

pectively. Increase in organic content surpassed the increase in

inorganic content, which resulted in lower percentages of ash

content (Table 1).

Pond-wise variation of amino acid composition and

chemical score of protein of mixed zooplankton are given in

Table 2. Nine essential amino acids were present in mixed

zooplankton. In most cases slightly over 50% by weight of the

Fig. 1. Monthly variation of SAFA a PUFa content in mixed zooplankton form different ponds.



7/15

total amino acids were essential amino acids. In general no

seasonal changes were found among individual amino acids

of zooplankton from different ponds but significant differ-

ences (Pb0.05) were present pond wise (Table 2). Chemical

score based on the requirement of different carp species

(Mohanty and Kaushik, 1991; Ravi and Devaraj, 1991; Mur-

thy and Varghese, 1998) revealed that methionine was limit-

ing for rohu (Labeo rohita), catla (Catla catla)and common

carp (Cyprinus carpio) in mixed zooplankton from all the

ponds. The quantity of phenylalanine was also less in respect

of carp requirement. Threonine was deficient for catla in the

amino acids of mixed zooplankton from all the ponds. How-

ever for mrigal (Cirrihinus mrigala) and catla it was deficient

from pond 1 and pond 5.

Fig. 2. Monthly variation of Ca, P and Fe content in mixed zooplankton form different ponds.



8/15

Selected fatty acids composition of mixed zooplankton is

indicated inTable 3. Nearly all the fatty acids found in mixed

plankton were straight chain molecules with even number of

carbon atoms.The content of total saturated fatty acids (SAFA),

mono unsaturated fatty acids (MUFA) and polyunsaturated

fatty acids (PUFA) ranged from 6481%, 715%, 620% of

total fatty acids. Among the saturated fatty acids myristic acid

(14:0), palmitic (16:0) and stearic acid (18:0) were present in

the plankton of all the ponds, and 16:0 was the dominant fatty

acid. Among the unsaturated, linoleic acid (LA, 18:2 n-6) was

present in the plankton of all the ponds. Whereas linolenic acid

(LNA, 18:3 n-3) was present only in the plankton of pond 1, 2

and 3, MUFA such as palmitoleic acid (16:1 n-7) andoleic acid,

(18:1 n-9) were present as 0.10 to 14.1% of total FA. Seasonal

Fig. 3. Monthly variation of Mn, Zn and Cu content in mixed zooplankton form different ponds.



9/15

variation in FA composition (Fig. 1) of zooplankton was pre-

sent in all the ponds irrespective of plankton composition

(Table 5). The percentage of SAFA increased appreciably

during June to September (water temperature 35 C) compared

with NovemberJanuary (18 C) and MarchApril (23 C)(Fig. 1). Contrary to this, the percentage of total PUFAs was

greater during NovemberJanuary. The PUFA were at their

lowest levels at 23 C.

The content of ascorbic acid in zooplankton from different

ponds ranged from 15 to 40.01g/g DM. Although pond wise

variation in the amount of ascorbic acid was present (Table 1),

no seasonal trend (apart from an increase ascorbic acid in

zooplankton of pond 4) was observed. No enzyme activity of l-

gulonolactone -oxidase could be detected in zooplankton.

Among the lipid soluble vitamins, average value of vitamin A

(retinol) was 13.6163.95 g/gm DM while vitamin E (total

tocopherol) averaged 218348 g/g DM. Vitamin A contentof zooplankton from all the ponds differed significantly (Pb

0.05) from each other. The analysis revealed that vitamin E had

a strong correlation (r1=0.72, r2=0.88, r3=0.82, r4=0.86,

r5=0.36, r6=0.88) with the lipid content of zooplankton. The

vitamin E content significantly varied (Pb0.05) pond wise

except for ponds 1, 4 and ponds 2, 3, respectively. Also the

vitamin E content was inversely proportional with that of

rising temperature.The amylase, protease and lipase activity of mixed zoo-

plankton ranged from 100226.1 g maltose/mg protein/h,

6.217.92 g leucine/mg protein/h, 25.8239.1 g -naphthol/

mg protein/h, respectively (Table 1).

Mineral and trace element composition is presented in

Table 1. Among the minerals P, Ca, and trace elements Fe, Cu,

Zn and Mn content were determined. Monthly variation of

mineral content is depicted inFigs. 2 and 3.

Different water quality variables prevailing in the ponds

during the study period are presented in Table 4. The water

temperature varied between 1835 C with minimum recorded

in DecemberJanuary and maximum in MayJune reflectingseasonal impact. No distinct trends in other physicochemical

parameters were discernible. Variation in population (%) and

average dryweight of zooplankton in differentpondsis presented

Table 5

Variation in population (%) and average dry weight of zooplankton in different ponds

June July Aug. Sept. Oct. Nov. Dec. Jan Feb Mar. Apr May Av. dry wt. mg/L

Pond-1 Cladoceran 71 80 73.9 74.6 74.0 77.3 76.7 70.7 69.2 81.3 74.0 59.1 4.2

Copepod 18.6 10.0 20.8 17.9 20.0 12.2 10.0 10.3 23.2 8.9 12.6 32.6

Rotifer 10.4 10.0 5.2 6.8 6.1 10.5 12.8 18.9 7.4 9.9 13.4 8.6

Pond-2 Cladoceran 92.3 67.9 62.8 59.7 63.6 60.8 69.6 66.4 57.6 68.5 63.6 80.2 4.2Copepod 4.8 11.9 26.2 25.7 23.3 15.0 21.4 14.2 35.7 21.5 12.5 8.1

Rotifer 2.9 20.2 11.0 14.6 13.1 24.2 9.0 19.4 6.7 10.0 23.9 11.7

Pond-3 Cladoceran 82.7 60.0 73.5 66.9 60.7 60.3 62.1 50.8 51.5 72.0 69.4 69.9 3.7

Copepod 8.0 36.2 14.7 21.9 22.3 27.5 19.4 33.0 30.1 18.0 20.7 13.0

Rotifer 9.3 3.8 11.8 11.2 17.0 12.2 18.5 16.2 18.4 10.0 12.0 7.1

Pond-4 Cladoceran 65.8 79.2 63.2 77.5 77.4 69.8 74.3 71.8 54.9 78.0 71.8 71.0

Copepod 26.1 10.1 22.6 15.5 19.0 18.9 19.7 78.8 27.8 11.7 11.7 24.0 1.8

Rotifer 8.1 10.7 14.2 7.0 3.5 11.3 5.9 9.7 173 10.3 10.3 5.1

Pond 5 Cladocerans 61.0 43.3 63.7 86.6 71.5 49.5 47.8 66.0 77.9 54.0 68.4 78.7 1.2

Copepods 33.9 13.4 6.7 6.7 13.3 42.1 46.79 24.9 10.2 28.2 23.6 18.5

Rotifer 5.1 43.3 29.6 6.7 15.2 8.4 5.3 10.1 11.1 17.8 8.0 2.8

Pond-6 Cladocerans 52.2 81.3 70.5 49.7 53.9 51.7 63.7 40.2 65.0 70.6 65.9 63.9 2.3

Copepods 40.6 8.6 12.6 42.2 32.9 42.9 10.6 31.6 21.5 14.6 27.1 20.1

Rotifer 7.2 10.1 16.9 8.1 13.2 5.4 26.7 28.3 13.5 15.8 7.0 16.0

Table 4

Variations of water quality parameters in different ponds

Pond-1 Pond-2 Pond-3 Pond-4 Pond-5 Pond-6

Temp (C) 1835 18.235.2 18.335.0 18.134.9 1835.1 1835

pH 7.17.3 7.07.3 7.27.3 7.17.3 7.37.4 7.10.4

Free CO2(mg/L) 1

6.2 1.6

8.0 1.8

6.0 1.0

8.0 2.0

8.0 1.8

8.0Dissolved oxygen (mg/L) 3.15.8 2.585.5 2.345.3 2.886.1 3.25.9 3.885.8

Alkalinity (mg CaCO3/L) 85101 89103 90100 88100 87100 86100

Total ammonia nitrogen (mg/L) 0.160.35 0.110.40 0.120.29 0.140.33 0.180.40 0.160.28

Nitrite nitrogen (mg/L) 0.080.18 0.060.12 0.030.15 0.020.09 0.020.09 0.010.09

Nitrate nitrogen (mg/L) 0.230.59 0.250.48 0.210.60 0.180.48 0.240.57 0.200.58

Phosphatephosphorus (mg/L) 0.160.19 0.110.23 0.150.20 0.100.22 0.150.24 0.170.30



10/15

inTable 5. Among the different groups of zooplankton Clado-

cerans were dominant because of their rapid proliferation. The

species present in the ponds were Moina (Moina dubia), Daphnia

(Daphnia lumholtzi,Daphnia carinata); Cyclops (Mesocyclops

hyalimus, Mesocyclops leuckarti); Diaptomus (Heliodiaptomus

viddus, Neodiaptomus handeli); Rotifer (Brachionus), and the

phytoplankton were Chlorella vulgaris, Volvox sp., Cyclotella

glomerata,Navicula,Cryptocyphala,Pediastrum, Oscillatoria,

Melosira, andAnabaena.

4. Discussion

Data on the chemical composition of mixed zoo-

plankton presented above provide basic information on

the nutritive potential of mixed zooplankton. The dry

matter content however was lower than what was re-

ported earlier (Yurkowski and Tabachek, 1979; Wata-nabe et al., 1983; van der Meeren et al., 2001). This was

because the plankton consisted of cladocerans that

contained less dry matter compared to other zooplank-

ton such as copepods (van der Meeren et al., 2001). This

might limit the required nutrient availability to the fish

larvae, emphasizing the essentiality of formulated feeds.

The protein content in D. carinata and Moina austra-

liensisis 54.34% and 64.80%, respectively (Kibria et al.,

1999) and inDaphniasp. it is reported to be 49.7070%

(Yurkowski and Tabachek, 1979; Watanabe et al., 1983)

whereas for Moina sp. it varies between 59.00 and

77.85% (Tay et al., 1991). The protein content of naturalzooplankton dominated by Temora longicornis varies

from 31% to 54% (Helland et al., 2003). Present in-

vestigation showed that the protein content of mixed

plankton varied from 7380% DM which is somewhat

higher than reported earlier (Tay et al., 1991) which

could be due to analytical methods used. So mixed

zooplankton might serve as a good source of protein for

Indian major carp larvae and early juveniles as well as

other freshwater fish larvae because larval fish generally

have high demand for dietary protein due to rapid

growth rates and extensive catabolism of amino acidsfor production of metabolic energy (Srivastava et al.,

2006). The total protein content of some copepods,

Eurytemore affinis, Centropages hamatus and Aeastia

grani from the Svartatjern lagoon of Norway was

quite stable, around 38% of DM (van der Meeren et al.,

2001). Present study showed the variability of protein

content in zooplankton from pond to pond but not much

variation within a pond. Though the species composi-

tion in the zooplankton population of different ponds

remained the same the relative abundance of different

zooplankton varied remarkably which might have

caused such variation in protein content.

The amino acids composition of mixed zooplankton

from different ponds had a relatively similar essential

and non-essential amino acids composition and the re-

lative amount was higher than previously reported (Yur-

kowski and Tabachek, 1979; Watanabe et al., 1983;

Kibria et al., 1999), which may be due to the mixedcommunity of zooplankton. Amino acid profile of plank-

ton is generally genetically programmed than diet re-

lated. Different kinds of food did not have any impact on

amino acid compositions of rotifers (Frolov et al., 1991).

Amino acid analysis byKibria et al. (1999)revealed that

both D. carinataand M. australiensis contained appre-

ciable levels of both essential and non-essential amino

acids for fish however, values of some essential amino

acids in D. carinata and M. australiensis were lower

than those previously cited forDaphniasp. (Yurkowski

and Tabachek, 1979; Hepher, 1988) and Moina sp.(Hepher, 1988). Artemia nauplii of different origin had

different amino acid profiles (Watanabe et al., 1983). But

the essential amino acid content of mixed zooplankton in

the present study could meet the general fish require-

ments particularly of carp (Table 2). But methionine was

deficient in the zooplankton of all the ponds, revealed

from CS(Table 2), which is similar with the other studies

(Yurkowski and Tabachek, 1979; Watanabe et al., 1983;

Srivastava et al., 2006).

Lipid content in mixed zooplankton from 6 ponds

varied from 10.79 to 14.55% DM and was inversely

related to water temperature, which is in agreement withthe findings ofJana and Manna (1993). The lipid con-

tent in freshwater zooplankton is known to have consi-

derable importance (Vijverberg and Frank, 1976) and

might be influenced by seasonal succession of phyto-

plankton species or source of food fed zooplankton

(Proulex and de la Nove, 1985; Kibria et al., 1999).

Watanabe et al. (1983) analysed various zooplankton

with Daphnia containing 13% and Moina 1227%

lipids whereas inD. carinataand Moina australiensisit

ranged from 7.297.73% (Kibria et al., 1999). Our

results also corroborated the above findings and alsocould meet the lipid requirement of carp which ranges

from 69% (Mukhopadhyay and Kaushik, 2001). The

percentage of SAFA increased remarkably during May

to July when the water temperature was 35C compared

with SeptemberOctober and MarchApril (23 C).

These differences in the levels of SAFA as the tempe-

rature decreases, were due to the decreased content of

16:0, which decreased by about half as the temperature

decreased from 35 C to 18 C. Contrary to this, the

percentage of total PUFA were greater at 18 C (23%)

compared to 35 C (10%). The PUFA were at their

lowest levels at 23 C. Substantial variation of fatty acid



11/15

composition in zooplankton along with seasonal varia-

tion of temperature was presumably an adaptation for

maintaining constant membrane fluidity. Water temper-

ature affects the fatty acid composition of algae and light

intensity affects the production of fatty acids (Tatsuzawa

and Takizawa, 1995), for which there was a seasonalchange in fatty acid composition of mixed zooplankton.

Our result showed close similarity with the finding of

Nanton and Castell (1999) who found significantly

higher content of PUFA at 6 C compared with 20 C,

but at intermediate temperature (15 C) it was lowest

where as MUFA were at their highest levels at 15 C in

both Amonardia and Tisbe, perhaps making up for the

lower levels of PUFA at this temperature. However, the

zooplankton was low in n-3 fatty acids, particularly of

LNA in some ponds and DHA and EPA in all ponds

which might have the similarity in the fatty acid patternof these herbivorous zooplankton and its phytoplank-

tonic food (Claus et al., 1979) containing LA and LNA

not EPA and DHA. However, freshwater algae unlike

marine algae generally have 18:3 n-3 and not 20:5 n-3

and 22:6 n-3 as their principal PUFA, in addition to 18:2

n-6 (Bell et al., 1994). It was noted thatDaphniasp. was

low in DHA andMoinasp. in both DHA and EPA. The

fatty acid composition ofB. plicatilis closely reflected

the fatty acid composition of the microalgae consumed

as food (Frolov et al., 1991) although biosynthesis rate

of PUFA de novo by B. plicatilis was rather slow

(Lubzens et al., 1985). Despite the different fatty acidcontent of microalgae, Scenedesmus abundans, Mo-

noraphidium minitum and Chlorella vulgaris, cultural

B. calyciflorusutilizing these algae was able to produce

its own lipid and fatty acid content to satisfactory levels

both qualitatively and quantitatively (Isik et al., 1999).

But in case of crustaceans fatty acid synthesis is gene-

rally low in Daphnids (de Lange and Arts, 1999). The

MUFA such as palmitoleic acid (16:1 n-7) and oleic acid

(18:1 n-9) in the mixed zooplankton might be syn-

thesised from the corresponding SAFA by the action of

9 desaturase. It has been suggested that harpacticoidcopepods contained sufficient amounts of the C18 to 20

and C20 to 22 elongase, and 6, 5 and putative 4

desaturase enzymes to be capable of this conversion

(Norsker and Stottrup, 1994; Nanton and Castell, 1998).

This desaturating ability a characteristic of the harpacti-

coid class could be an adaptation to a long-chain EFA-

poor benthic environment when compared to the pelagic

calanoid copepods. A calanoid copepod, Paracalanus

parvus also appears to have some limited ability to

elongate and desaturate 18:3 n-3 to the longer chain n-3

HUFA (Nanton and Castell, 1998). Our results showed

that the calanoid copepods as well as other cladocerans

and rotifers present in the mixed zooplankton were in-

capable of the above discussed coversions which could

decrease its EFA value for larval fish as well as adults.

Absence of l-gulonolactone -oxidase activity con-

firmed the inability of these zooplankton to synthesize

ascorbic acid (AA) de novo. So the zooplankton mustdepend on an exogenous supply. In this study the

presence of ascorbic acid in zooplankton from different

ponds demonstrated that the occurrence and variability

of AA in the organism was related to the feeding status

of the species (Poulet et al., 1989; Hapette and Poulet,

1990; Merchie et al., 1995). In the pond zooplankton

were grown feeding on phytoplankton. Generally the

phytoplankton (different micro algal sp) are rich source

of AA, but show a considerable variability among the

different species and different phase of life cycle (Mer-

chie et al., 1995) as well as environmental conditionsuch as pH, CO2, light intensity, photoperiod, temper-

ature, cells physiological response to inorganic macro-

nutrients such as nitrate, and concentration of trace

metals (Brown and Lavens, 2001; de Castro Araujo and

Garcia, 2005). Studied environmental condition proba-

bly contributed less production of ascorbic acid in the

phytoplankton which was reflected in zooplankton. The

National Research Council (1993) recommends 25

50 mg AA/kg diet as a requirement to secure an optimal

performance of juvenile fish. Dabrowski (1990) sug-

gested that the metabolic rate is the primary factor re-

gulating the AA requirements. Therefore, larval fish,displaying a relatively faster growth and metabolism

than juveniles and adults might need high dietary AA

levels to sustain optimal growth and physiological con-

dition (Dabrowski, 1990). Six hundred fifty to 750 mg/

kg diet of AA has been recommended for Cirrhinus

mrigala during early growth (Mahajan and Agrawal,

1980). This study clearly indicated the impairment of

the zooplankton to fulfil the vitamin C requirement for

fish larvae and early juveniles. The content of vitamin C

in zooplankton from ponds 1, 2, 3 and 4 was below the

recommended level of NRC and the content of vitaminC from other ponds was within the recommended level.

So additional dietary source of vitamin C might be

mandatory for healthy growth and development of carp

larvae. Vitamin A content varied between 13.61

63.95 g/gm DM. The requirement of dietary vitamin

A forL. rohita has been recommended as 606.6 g/kg

dry feed (Rangacharyulu et al., 1999) and 121.21

606 g /100 g dry diet for young carp. The values

obtained in the present study were much higher than the

reported values in other zooplankton (Moren et al.,

2004). Less work has been done on the vitamin E re-

quirement of carp larvae. The vitamin E supplementation



12/15

at the minimal requirement level as defined by NRC

(1993) is 50 g/kg feed. Assessed data were 228

333 g/g DM which is more than the NRC recom-

mended value. The broodstock diets containing 60 g

ascorbic acid (pure ascorbic acid powder)/g dry feed and

300 g vitamin E/g dry feed are sufficient to ensureproper reproduction and offspring quality (Mukhopad-

hyay et al., 2003).

Information about mineral and trace element require-

ment in Indian major carp is limited. Relatively few data

exist about the influence of minerals in prey on larval

feeding. Some data exist for the macro mineral and trace

element compositions of Artemia, which is variable

depending on stock (Rabin and Gatesoupe, 2001). In the

present study the concentration of Ca in zooplankton

varied from 1.052.99 mg/100 g, which is less than the

requirement of common carp 0.3

3.0 g/kg (Steffens,1989; Kaushik, 2001). The bioavailability of phospho-

rus from zooplankton is not known. The phosphorous

content 0.240.94 mg/100 gm DM of zooplankton was

much less than common carp requirement (600 mg/

100 g) (Kaushik, 2001). The concentration of iron in the

zooplankton of pond 2 was highest among the others.

Iron level in the zooplankton was lower than the level

used in prepared feed and the requirement reported for

common carp (200 mg/kg) (Steffens, 1989). High levels

of dietary iron for fish have recently been questioned

with regard to negative health effects in relation to

pathogen bacteria development (Lie et al., 1997). Itmight be prudent to look into ways to maintain the iron

concentration in zooplankton. The zinc concentration of

the zooplankton was found to be 0.022.88 mg/100g

DM. This concentration range is not much less than the

requirement found for fast growing fish (Maage and

Julshamn, 1993) as well as common carp (1530 mg/

kg) (Steffens, 1989; Lim et al., 2000). As absorption of

dissolved zinc (occurs mainly across the gills) is low in

freshwater fish (Kaushik, 2001), this dietary source has

great relevance in freshwater aquaculture. Manganese

content of zooplankton varied from 0.020.49 mg/100 g, which is also less than the requirement of com-

mon carp (1213 mg/kg) (Steffens, 1989). Dietary cop-

per level of 3 mg/L improved growth in common carp

fry (Ogino and Yang, 1980). The concentration of cop-

per in mixed zooplankton varied from 0.0010.181mg/

100g DM.

The role of zooplankton proteases as activators of

fish zymogen might be of importance because of the

direct contribution of proteolytic activity to the autolytic

processes of food organisms (Dabrowski and Kaushik,

1984). Kurokawa et al. (1998) demonstrated that pro-

teases derived from live food had only a small contri-

bution to the enzymatic activity measured in sea bass

and sardine larva, respectively. The role of zooplankton

amylase and lipase in larval nutrition might also be

significant. The present study revealed that zooplankton

from different ponds were a good source of amylase,

protease and lipase. The contribution of lipolytic en-zymes to the total lypolytic process was less than 6.5%

of the total enzyme activity of stripped bass (Morone

saxatilis) during ontogenic development (Ozkizilcik

et al., 1996). The role of dietary enzymes in larval

digestion, the characterization and quantitative estima-

tion of digestive enzymes from zooplankton might be

necessary to determine the enzyme input from live food

to fish larvae.

In the present study though the fertilization condi-

tions were same in all the ponds, the phytoplankton

species as well as the zooplankton population variedgreatly among the ponds, which might resulted in the

pond wise variation of chemical composition of zoo-

plankton. Particular sequence of changes in temperature,

photoperiod and light intensity might alter the propor-

tion of the individual constituents of phytoplankton, the

main food of zooplankton. However zooplankton in

ponds has two sources generally: allochthonous popu-

lation from the water used to fill and the autochthonous

population from resting form in the sediment. In the

present study though zooplankton were present in water

source it was the same for all the ponds. Therefore only

autochthonous population from resting form in the sedi-ment developed and might have added in the variation in

zooplankton population (Milstein et al., 2006). Frolov

et al. (1991)found a positive correlation between lipid

and fatty acid content, of rotifer and content of these

compounds in their food. However, not all biochemical

components (like carbohydrate, amino acid) changed to

the same degree since, in the course of evolution, an

increase in the conservation of some groups of sub-

stances and stability of others had taken place, resulting

in high stability of organisms as a whole. However,

apart from environmental conditions and food, harvest-ing techniques, and sample preparation are vital factors

that may affect the nutrient composition (Millamena

et al., 1990). The nutritional quality in Artemia also

varied considerably with the variation of the geograph-

ical origin (Leger et al., 1986; Han et al., 2001), to

differences among different batches of cysts from the

same origin, and to the methods of analysis. Among the

water quality variables variation in temperature reflected

seasonal impact. Certain degree of variation in other

water quality variables was registered between different

ponds, and between the different samplings during the

course of investigation. The results of such variations



13/15

were obvious in field conditions. The inorganic nut-

rients such as NO2N, NO3N, and PO4P values also

showed no particular trend during study period, attri-

butable to the intermittent fertilization practices (Jena

et al., 1998).

5. Conclusion

Freshwater mixed zooplankton contained a high

amount of protein (although deficient in methionine),

vitamin A and vitamin E and low in vitamin C with no

ascorbic acid synthesizing capacity de novo. Although

the lipid content was high, it was low in some essential

fatty acids like LNA, EPA and DHA, which could be

improved through different fatty acid enrichment stra-

tegies according to the requirement of the desired spe-

cies. Appreciable amount of digestive enzymes waspresent in mixed zooplankton that could be utilized

during ontogenesis for better nutrient management

through increasing nutrient digestibility of the larvae

for healthy stockable seed production. Substantial dif-

ferences were found in the mineral composition of the

zooplankton studied to the requirement of fish. This

information may have nutritional implication in culture

of fish larvae and raising of juveniles so that provision of

the prepared feed should be to complement what a pond

ecosystem provides rather than to supplement with a

diet of generic nature chosen empirically.

Acknowledgements

The authors are grateful to the Director, Central

Institute of Freshwater Aquaculture, Bhubaneswar for

providing facilities for carrying out the work. Financial

assistance from Indian Council of Agricultural Research

as Senior Research Fellowship to the senior author is

thankfully acknowledged.

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