The study investigates the impact of different diet categories on muscle growth dynamics in the freshwater prawn, Macrobrachium rosenbergii, over a 210-day period. Four diet categories were formulated using various floral components, with significant findings indicating that the diet with green algae produced the largest average muscle fiber surface area. These results highlight the relationship between diet composition and muscle growth, suggesting potential avenues for improving aquaculture practices.

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Int J Adv Pharm Biol Sci Vol.4, Issue 3, 01-07 KUNAL et al.
ISSN 2249 –8966
Research Article Available online http://www.ijapbs.in
IMPACT OF DIFFERENT DIET CATEGORIES ON MUSCLE GROWTH
DYNAMICS OF FRESHWATER PRAWN, MACROBRACHIUM ROSENBERGII
Mondal Kunal*1, Bhattacharyya Subhra B2, Mitra Abhijit2, Das Madhusudan3
1 Department of Marine Science, University of Calcutta, 35 B.C. Road,
Kolkata- 700019, India
2 Department of Marine Science, University of Calcutta, 35 B.C. Road,
Kolkata- 700019, India
3 Department of Zoology, University of Calcutta, 35 B.C. Road,
Kolkata- 700019, India
Received: 10th May 2014, Revised and Accepted: 24th May 2014
*Corresponding author: kunalmondal1985@yahoo.com
INTRODUCTION
Prawn growth is regulated by the fundamental process of molting associated with
complex biochemical activity of specific proteins that helps withdrawal of exoskeleton
from the carapace during muscle growth and development. "Muscle cellularity" is the
key indicator mechanism behind the skeletal growth of a particular organism that
INTERNATIONAL JOURNAL OF ADVANCE PHARMACEUTICAL AND
BIOLOGICAL SCIENCES Vol. 4, Issue. 3, JULY-SEPTEMBER 2014
ABSTRACT
Artificial diets were formulated with three different categories of floral components viz. green
algae Enteromorpha intestinalis (ENT diet category), salt-marsh grass Porteresia coarctata
(POT diet category) and mangrove litter (ML diet category), mixed with fishmeal at a level of
5%. Simultaneously diet with 0% fishmeal replacement was also formulated (control diet
category). The study aimed to investigate the impact of diet categories on muscle growth
dynamics of freshwater prawn, Macrobrachium rosenbergii reared in farm condition. A 210
day growth trial was conducted with the test species, histological observations reveal marked
cytomorphological changes (hypertrophy) in abdominal muscle of the organism when
compared between initial and final stage of growth in relation to each diet category. Change
was identified on the basis of muscle fibre enlargement in terms of surface area. ANOVA
results confirm significant variations (p<0.05) in average fibre surface area of prawn muscle
exposed to different diet categories. The fibre surface area varied as per the sequence of
prawns fed with ENT diet category (16.80 ± 12.44 mm2) > control diet category (14.40± 5.37
mm2) > POT diet category (13.0 ± 3.39 mm2) > ML diet category (9.50 ± 4.10 mm2)
respectively. Thus muscle growth pattern of a particular species is clearly related to the kind
of diet provided and its chemical composition as evidenced from the present experimental
study.
Keywords: Floral components, diet categories, M. rosenbergii, muscle fibre, hypertrophy.

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involves the contrasting balance of hyperplasia and hypertrophy of the muscle fibre[1-3].
This delicate balance may sometimes vary depending upon various internal and
external factors. Research findings reveal that diet is one of the significant extrinsic
environmental factor governing the muscle growth of fish thereby causing such
variation in fibre size and number[4-6]. Muscle fibre distribution during growth have
altered the production of commercially important fishes like atlantic salmon[7], arctic
charr[8] and rainbow trout[9, 10]. The giant freshwater prawn M. rosenbergii is also an
important culture species across the globe and its commercial productivity is more
dependent upon the category of diets provided to achieve the target features like overall
growth and body weight. Very little information about muscle growth pattern and its
magnitude of variation upon diets is reported in case of this particular aquatic species.
Therefore the present experiment is an attempt to investigate the impact of different
formulated diet categories on muscle growth dynamics of the species reared in farm
condition.
MATERIALS AND METHODS
Floral components and diets: Live and healthy floral components (E. intestinalis, P.
coarctata and mangrove litter) were collected from the intertidal mudflat in the
mangrove belt of Indian Sundarbans (21o52'35.7'' N latitude and 88o11' 55.0'' E
longitude) during low tide condition. All the collected plant materials were washed in
ambient water and then with freshwater to remove epiphytes and other extraneous
matter, dried in hot air oven under 55oC to preserve the biochemical constituents and
then finally processed. The floral components were mixed with fishmeal at a level of 5%
within the diets to meet the nutritional requirements of prawn (Table 1). The categories
of diets formulated are as follows:
ENT diet category= Diet incorporated with E. intestinalis powder
POT diet category= Diet incorporated with P. coarctata powder
ML diet category= Diet incorporated with powdered mangrove litter
Control diet category= Prepared with fishmeal
Table No. 1 Formulation of different categories of diets supplemented with floral
components at 5% level (% dry matter)
Growth trial: A growth trial was conducted at Kakdwip region of Indian Sundarbans
(21o52'35.7'' N latitude and 88o11' 55.0'' E longitude) in grow-out farm conditions. The
experimental facility consisted of triplicate ponds of each diet category. Prawn juveniles
of a mean initial weight of 2.5 gm were obtained from a local hatchery and acclimated to
INGREDIENTS CONTROL DIET
CATEGORY
ML DIET
CATEGORY
ENT DIET
CATEGORY
POT DIET
CATEGORY
Fish meal 35 30 30 30
Soybean oil cake 11 11 11 11
Mustard oil cake 11 11 11 11
Rice polish 23 23 23 23
Wheat flour 16 16 16 16
Oyster shell dust 2 2 2 2
Shark oil 2 2 2 2
Mangrove litter - 5 - -
E. intestinalis - - 5 -
P. coarctata - - - 5

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the conditions prior to 210 day growth trial. Stocking at a density of 2 individuals/m2
was done in all the experimental ponds. Dietary treatments were randomly assigned to
the ponds and feeding on the respective diet categories was initiated.
Histological sampling: Samples were collected at 60 DOC (initial) and 210 DOC (final).
Body weight were measured prior to sacrificing the animals. Muscle samples were
collected by removing the shell from the animals, tissue from second abdominal
segment was dissected and about 1.0 gm of tissue was fixed in Bouin's solution. [DOC=
days of culture].
Tissue processing and staining: The fixed tissues were washed in running tap water
and then dehydrated gradually by immersing in different concentrations of alcohol.
After dehydration, the tissue samples were immersed in cedar wood oil until they
became totally transparent. The transparent tissues were impregnated in xylene:
paraffin (1:1) mixture at 60oC for 1 hour and then transferred to fresh paraffin for
another 1 hour. The whole procedure was repeated twice. The tissues were embedded
in paraffin blocks and then sectioned into 4µ thickness with a microtome and mounted
on slides. After drying, the slides were immersed in xylene to remove the wax
completely. The deparaffinized tissue sections were rehydrated and stained with
haematoxylin and counter stained with eosin. The sections were mounted using
Distrene, Plasticiser, Xylene (DPX).
Measurement of muscle fibre surface area: Muscle cross-sectional images were
taken with the help of NIKON phase contrast microscope fitted with an image analyzing
system. The surface area enlargement of the fibre were measured using a 'Planimeter'
to quantify the degree of muscle growth in prawns from each diet category. The areas of
ten randomly selected fibre were measured per slide and the average was estimated. A
total of three slides were analyzed for each diet category.
Statistical Analysis: The collected data were finally subjected to one-way analysis of
variance (ANOVA). All calculations were performed with Web Agri Stat Package (WASP)
statistical software for Windows.
RESULTS
Muscle tissues were evaluated microscopically to identify the histological changes
related to the application of different categories of formulated diet. Cross-sectional
images showed distinct cytomorphological changes i.e. enlargement of muscle fibre, a
comparative account of which is presented (Fig. 1). In the groups of prawn fed with ENT
diet category, the average fibre surface area was 16.80 ± 12.44 mm2. Prawn groups fed
with POT diet category exhibited an average fibre surface area of 13.0 ± 3.39 mm2.
Similarly groups of prawn fed with ML and control diet categories showed an average
fibre surface area of 9.50 ± 4.10 mm2 and 14.40 ± 5.37 mm2 respectively. ANOVA results
also confirmed significant variation (p<0.05) in average fibre surface area and body
weight of the species (Table 2).

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Table No. 2 Variations in average fibre surface area and body weight of
M. rosenbergii fed with different diet categories
*Means bearing different superscript within a group differ significantly from each other
(p<0.05); values are means of three replicates.
Fig 1: Cross-sectional view of abdominal muscle tissue of M. rosenbergii at 60 DOC
and 210 DOC. H & E stain, ×40. Arrow signs indicate marked cytomorphological
changes i.e. enlargement of muscle fibre (mf) in (A) ENT diet category (B) POT diet
category (C) ML diet category and (D) control diet category.
60 DOC 210 DOC
A
B
C
PARAMETERS ENT DIET
CATEGORY
POT DIET
CATEGORY
ML DIET
CATEGORY
CONTROL DIET
CATEGORY
Average fibre surface
area (mm2)
16.80± 12.44a 13.0± 3.39c 9.50± 4.10d 14.40± 5.37b
Body weight (gm) 69.92a 57.07c 52.50d 62.95b
mf mf

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D
DISCUSSION
Muscle growth in fishes involves the combined action of hyperplasia (i.e. recruitment of
new fibre) along with hypertrophy (i.e. enlargement of existing fibre) which determines
the overall muscle mass. The data obtained thus indicates that there might be specific
contributors to muscle growth in M. rosenbergii fed with ENT diet category. These seem
to play a key role in boosting growth by regulating body weight and muscle plumpness.
Histological observations reveal hypertrophic muscle growth in the organism which
occurs by fusion of existing fibres, thus adding new nuclei but varies accordingly with
the respective diet categories. The probable mechanism for such hypertrophic growth
may be attributed to (i) presence of chemical compound dimethyl sulfonyl propionate
(DMSP) within this particular algal group that provides improved growth performance
by accelerating better consumption[11, 12] (ii) presence of any natural growth promoting
compound that still remains unidentified in this algal species and (iii) differential
expression of growth-related genes due to a particular diet composition. However
further studies are required to unlock the triggering factors as well as the effect of
diets/ nutrients on muscle gene expression, which is actually a specialized domain of
research i.e. 'aquaculture nutrigenomics'. Nevertheless ambient environmental
condition of the rearing system also has a profound influence on the growth of the
species which needs to be considered in the present experimental study.
A similar work has been reported previously where M. rosenbergii stocks collected from
different natural reservoirs of Indian region when reared in hapa-condition exhibited
significant variation in diameter of muscle fibre[13]. Observations while working with
marine shrimp, Litopenaeus vannamei states that specific morphological changes were
observed in abdominal muscle associated with different stages of molt cycle[14].
Significant correlation was found between the fibre size in relation to age as well as
influence of environmental factors like diet in various fish species[15-21].
CONCLUSION
The present study provides a baseline information about the muscle growth dynamics
of M. rosenbergii exposed to different diet categories in captive condition and
accentuates the need for further extensive research. Variations in hypertrophic muscle
growth between Macrobrachium groups fed with four different diet categories opens up
the avenue of selective breeding programs by altering the muscle architecture and
consequently overall growth and body weight.
ACKNOWLEDGEMENTS
The authors wish to thank Department of Marine Science and Department of Zoology,
University of Calcutta for providing all facilities to carry out the present work. The

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authors acknowledge the financial help from Department of Science and Technology,
Govt. of India to carry out this work.
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