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J. Env. Bio-Sci., 2014: Vol.

28 (2): 293-298 ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

EFFECT OF ENRICHED MOINA AND MICROPARTICULATE DIET ON FATTY ACID


COMPOSITION OF PENGBA (OSTEOBRAMA BELANGERI) LARVAE
R. Ramesh1#, Kiran Dube1, A. K. Reddy1, P. V. Rangacharyulu2 and G. Venkateshwarlu1
Central Institute of Fisheries Education, Panch Marg, Off Yari Road, Versova, Andheri (West), Mumbai - 400 061,
Maharashtra, India1
Regional Research Centre, CIFA, Fish seed farm, Penamaluru, Poranki P.O., Vijayawada - 521 137, Andhra Pradesh, India2
email: rathodcifa@gmail.com#

Received: 25-09-2014 Accepted: 29-10-2014


In order to evaluate the effect of enriched Moina and microparticulate diet with different combinations on the tissue fatty acid
composition pengba larvae was studied. The feeding trial was conducted for the larvae using seven experimental diets consist of
Moina (enriched with algae and cod liver oil) and combination of enriched Moina and micro particulate diet. A total of thirty one fatty
acids were identified by GC-MS encompassing saturated, mono unsaturated, n-6 and n-3 poly unsaturated fatty acids. The n-6 and
n-3 PUFA showed a dramatic proportional increase in larval tissue relative to its proportional composition in both the live and
formulated diets. Dietary levels of the n-6 and n-3 fatty acids were similar among diets-4, 5, 6 (enriched Moina with cod liver oil and
algae). However, these fatty acids levels in the larvae were significantly higher in larvae fed combination of enriched Moina and
microparticulate diet (Diet-5 and Diet-6) compared to larvae fed other diets. Proportional increases of AA, EPA and DHA (20:4n-6,
20:5n-3 and 22:6n-3 respectively) in larvae relative to dietary HUFA, suggest important physiological roles for these fatty acids. The
increase in n-3/n-6 ratio vice versa demonstrated that larvae may require the higher EPA and DHA in their first feeding. The effect
of different feeding regimes of respective diets fed to pengba larvae based on fatty acid composition were clearly established by
principal component analysis.

Live feed has become the most important secondary producers the State Fish of Manipur, Northeastern states of India. The
in aquatic food webs, and has been extensively used as high fish can be considered as potential fish species for
quality natural food in hatcheries for the maintenance or diversification of carp culture due to its better taste and meat
propagation of early larval stages of fishes for the past few quality. As part of studies on development of breeding and
decades1. Moina has recently become a potential live feed, production technology of pengba, the evaluation of nutritional
with respect to its availability, in most of the natural water requirement of this species needs to be determined. So far no
resources and has a high nutritional value. This freshwater attempts were made to improve the fatty acid composition of
zooplankton is classified as a cladoceran and it has been pengba larvae with respect to live feed enrichment and artificial
documented as a potential live feed to substitute the use of diets for their better growth and survival. In this background,
the more expensive Artemia and other seasonal zooplankton the study was conducted to emphasize the effect of co-feeding
such as copepods, in larviculture2. on fatty acid composition of larval pengba with Moina and
microparticulate diet.
In many hatcheries, common enrichment practices to enrich
fatty acids composition in Artemia and other live feeds such MATERIALS AND METHODS
as rotifers consisted of microalgae, microalgae paste, and Moina culture and enrichment
various marine fish oils3. Das et al. found that the nutritional
value of Moina could be similarly enhanced by enrichment Mass culture of Moina and enrichment was followed the method
with various culture media as demonstrated in Artemia nauplii of Parakarma et al4. The composition of prepared emulsion for
and rotifer2. However, there is limited information, and no notable enrichment of Moina is given in Table -1.
study has been found on M. micrura enrichment, especially Experimental fish and feeding strategies:Pengba larvae
on the aspects of culture media selection such as micro algal (three days old post yolk sac) were obtained from hatchery
enrichment concentration and appropriate treatment time. upon successful breeding of pengba brooders. Total 2,100 one
Pengba, Osteobrama belangeri is a medium carp regarded as week old larvae (average weight 0.15±0.01 mg) were randomly
Effect of Enriched Moina and Microparticulate Diet on Fatty Acid (294)

[(Diet-1 (Moina%), Diet-2 (Moina + MD), Diet-3 (Moina enriched with cod liver oil), Diet-4 (Moina enriched with cod liver oil + MD),
Diet-5 (Moina enriched with algae), Diet-6 (Moina enriched with algae + MD) and Diet-7 (Microparticulate diet)].

Fig. 1. Bi-plot of distribution in fatty composition of larvae of pengba (Osteobrama belangeri)


fed with different feeding regimes

Table-1. Composition of emulsion media used for Moina enrichment

distributed into 21 smaller rectangular tanks (60cm height, libitum daily twice at 08:00 and 17:00 h. Uneaten feed was
74cm length, 44cm width) at the density of 100 larvae per siphoned daily to maintain optimum water quality. The water
tank. The feeding trial was conducted for 35 days with three temperature was recorded 22.2±2.8 °C and water quality
replicates of each treatment following a completely parameters such as Dissolved oxygen level was 6.2±0.66 mg/l,
randomized design. Fish larvae was fed with seven pH 7.2±0.50, alkalinity 152.5±4.56 mg/l, total hardness 300-
experimental diets namely Moina (Diet-1), Moina + MD (Diet- 400 mg/l and NH4 0.1 mg/l were recorded throughout the
2), Moina enriched with cod liver oil (Diet-3), Moina enriched experiment.
with cod liver oil + MD (Diet-4), Moina enriched with algae
(Diet-5), Moina enriched with algae + MD (Diet-6) and Total lipid and Fatty acid ethyl esters (FAME)
Microparticulate diet (MD) (Diet-7). The larvae were fed ad Total lipids of enriched and micro particulate diets were extracted
(295) Ramesh, Dube, Reddy, Rangacharyulu and Venkateshwarlu

Table-2. Fatty acid composition (% of total fatty acids) of different diets fed to pengba larvae

[ND: Not Detected; CLO:Cod liver oil; MD: Microparticulate diet]


Effect of Enriched Moina and Microparticulate Diet on Fatty Acid (296)

Table-3. Fatty acid composition (% of total fatty acids) of pengba larvae fed with different
feeding regimes

[ND: Not Detected]


(297) Ramesh, Dube, Reddy, Rangacharyulu and Venkateshwarlu

following the method of Folch et al5. The AOAC method was PUFA identified, accounting 0.32-3.66%, 0.34-2.68% and 0.32-
followed to esterify the lipid extract6. Fatty acid methyl esters 1.36%, respectively.
(FAME) were analyzed using a Shimadzu QP2010 Quadruple In the present study, the fatty acid composition of fish larvae
Gas Chromatography Mass Spectrometer (GCMS) equipped is directly influenced by the different dietary treatments. The
with a Carbowax (30 m x 0.25 mm ID; 0.25- μ m film thickness) fatty acid composition of fish in general is known to be
capillary column (Cromlab S.A.). Helium was used as the carrier influenced by the fatty acid composition of dietary lipid7. Further,
gas. The values of fatty acids are present in peak area tissue fatty acid composition is affected by size or age of
percentages of total identified fatty acids. Statistical animals8, food deprivation9 and non-dietary factors including
analysis.The multivariate data on fatty acid composition of environmental temperature10. The presence of high amount 16:0
larvae subjected to different feeding regimes with enriched and 18:1n-9 fatty acids in the present study supported by
Moina and microparticulate diets were analyzed by principal Sargent et al, who proposed that 16:0 and 18:1 n-9 are the
component analysis (PCA) using software Unscrambler most preferred fatty acids for the mitochondrial β -oxidation
(Version 9.5, CAMO, Norway). and thus the source of metabolic energy in fish11. The results
RESULTS AND DISCUSSION are in agreement with Faulk and Holt found that enriching
rotifers and Artemia with live algae can enhance the HUFAs
Fatty acid composition of diets: The fatty acid composition content in the live feed and consequently improve the growth
of Moina (enriched with cod liver oil and micro algae) and and survival of cobia larvae12.
microparticulate diet is summarized in Table-2. From the fatty
PUFAs in live organisms like microalgae are much more stable
acid data, total saturated fatty acids (SAFA), total
than in commercially available emulsion formulations. It is
monounsaturated fatty acids (MUFA), sum of n-6 and n-3 poly
significant that the levels of AA, EPA and DHA found in enriched
in saturated fatty acids (PUFA) and the ratios of n-3/n-6, n-6/
live microalgal cells more closely resemble larval natural diets.
n-3 were calculated. The SAFA level was highest in unenriched
In particular, the competitive interactions between AA and EPA
Moina (46.49%) and lowest (32.05%) was found in
are important in the formation of eicosanoids13. The present
microparticulate diet. MUFA level was highest (38.8%) in Moina
study the AA levels are very less in the body tissues which
enriched with cod liver oil, whereas lowest (30.87%) was
contrast to earlier studies conducted by Copeman et al in
observed in unenriched Moina. However, the highest n-6 and
yellowtail flounder where AA levels were increased in body
n-3 PUFA were found in microparticulate diet (22.64%) and
tissues in spite of lower AA level in the diet14. The ratio of n-6
enriched Moina (11.07%, CLO and 11.69%, algae) respectively.
and n-3 PUFA were ranged from 0.27 to 3.66 revealed that the
Fatty acid composition of larvae:The fatty acid profiles of importance of EPA and DHA concentrations, were influenced
larvae fed with different dietary feeding regimes were presented in the dietary treatments. High ratio of n-6 and n-3 PUFA were
in Table-3. In pengba larvae, the SAFA level was highest in found in larval tissues suggested that EPA and DHA contributed
Diet-1 (57%) and lowest (46.23%) was found in Diet-6. MUFA in their biological activities. These findings were supported by
level was highest (38.8%) in Diet-2 and lowest (30.87%) was earlier workers Watanabe et al. and Koven et al. proposed
observed in Diet-7. The highest n-6 and n-3 PUFA were found notably in marine fish species 15, 16.
in Diet-6 (10.22%) and Diet-7 (8.47%) respectively. The most
To gain better insight into the data structure, principal
common SAFA was dominated by palmitic acid (29.35-39.12%)
component analysis (PCA) was performed on the fatty acid
and stearic acid (4.12-6.21%). Among MUFA, oleic acid (18:1n-
composition of larval samples obtained at seven different
9) was the most abundant comprising of 18.13-33.27% of the
feeding regimes. The projection of variables and different diets
total fatty acids followed by palmitoleic acid (16:1n-7) which
onto the first two principal components was given in Fig. 1 in
formed 0.54-4.24%. Linoleic acid (18:2n-6) and Arachidionic
the form of bi-plot. The analysis explored in a two-principal
acid (20:4n-6) were the principal n-6 PUFA at a level of 1.04-
component (PC1 and PC2) model that described 71% of the
6.46% and 0.19-3.24% of total fatty acids respectively. Linolenic
total data variability. The pengba larvae fed with different diets
acid (18:3n-3), Eicosapentaenoic acid (EPA, 20:5n-3) and
were separated based on fatty acid composition and
Docosahexaenoic acid (DHA, 22:6n-3) were the major n-3
demonstrated that fatty acid profile of pengba larvae were clearly
Effect of Enriched Moina and Microparticulate Diet on Fatty Acid (298)

influenced by different feeding regimes. PC1, which explained 4. Parakarma, M.G.I.S., Rawat, K.D., Venkateshwarlu, G. and
44% of total information in the data, has clearly separated Reddy, A.K. (2009). J. Ind. Fish. Assoc., 36:9.
diet-5 and diet-6 from other diets. Further, it can be clearly 5. Folch, J.L., Sloane, M. and Stanley, G.H.S. (1957). J. Biol.
noticed from the bi-plot, that the pengba larvae fed with diet-5 Chem., 226: 497.
and diet-6 were rich in n-6 and n-3 fatty acids such as 18:2n- 6. AOAC (1995) In: Official Methods of Analysis of the
6, 20:2n-6, EPA, DPA and DHA, were found to be responsible Association of Official Analytical Chemists, AOAC,
for causing the differences among the samples. The present Arlington, VA.
study was supported by Singh et al., where the PCA showed
7. Torstensen, B.E., Lie, O. and Froyland, L. (2000). Lipids,
the differences in the fatty acid profiles of fish fry fed with
35:653.
different dietary groups enriched with live feed and formulated
8. Kiessling, A., Pickova, J., Johansson, L., Asgard, T.,
diets and was found to be responsible for causing the
Storebakken, T. and Kiessling, K. (2001). Food Chem.,
differences among the samples17.
73:271.
In conclusion, enrichment of Moina (algae and cod liver oil)
9. De Silva, S.S., Gunasekera, R.M., Collins, R., Ingram,
combination diets contained relative proportional of fatty acids
B.A. and Austin, C.M. (1997). J. Fish Biol., 50 (5): 992.
along with their favorable n-3/n-6 ratios which are directly
10. Bell, M.V., Henderson, R. J. and Sargent, J. R. (1986).
proportional to the larval tissue fatty acids. The EPA and DHA
Com. Biochem. Physiol., 83B: 711.
found in pengba larvae fed with various dietary treatments
assume that they played a significant role in their physiological 11. Sargent, J.R., Tocher, D.R. and Bell, J.G. (2002). In: Fish
activity. The co-feeding method was effective in improving the nutrition, Academic Press, San Diego, CA, pp. 181.
nutritional values of larvae for their better growth and survival. 12. Faulk, C.K., Holt, G.J. (2005). Aquaculture, 249: 231.
13. Harel, M. and Place, A.R. (2003). Comp. Biochem.
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