Food and Chemical Toxicology 47 (2009) 119123

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Food and Chemical Toxicology

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Elemental composition in mud crab Scylla serrata from Mahanadi estuary,

India: In situ irradiation analysis by external PIXE
Anil Mohapatra a,*,1, T.R. Rautray b, Ajit K. Patra c, V. Vijayan d, Rajeeb K. Mohanty e
a

Department of Environmental Sciences, Fakir Mohan University, Vyasa Vihar, Balasore-19, Orissa, India
Institute of Physics, Sachivalaya Marg, Bhubaneswar 751005, India
c
Institute of Minerals and Materials Technology, Bhubaneswar 751013, Orissa, India
d
Dept. of Physics, Valliammai Engineering College, SRM Nagar, Chennai 603203, India
e
Water Technology Centre for Eastern Region (ICAR), Bhubaneswar 751023, India
b

a r t i c l e

i n f o

Article history:
Received 6 May 2008
Accepted 15 October 2008

Keywords:
Crustacean tissue
Metal content
Bioaccumulation

a b s t r a c t
During the present study concentration of nine elements (K, Ca, Mn, Fe, Cu, Zn, Se, Br and Pb) in different
tissues of mud crab Scylla serrata from Mahanadi estuary, India were determined by the external PIXE set
up at Institute of Physics, Bhubaneswar, India. The study demonstrates the effectiveness of the technique
in analyzing both soft and hard tissue samples from marine organisms and opens the door for nondestructive, multi-elemental analysis of tissue samples with a very little sample preparation by direct
irradiation. This technique can be well utilized for analyzing the tissue samples for environmental, toxicological and nutritional purposes. The study also demonstrates the elemental concentrations from tissue samples of any crustaceans from Mahanadi estuary for the rst time. Sex based difference in the
elemental concentration of the mud crabs were marked, which may be related to the growth rate and
other biological activities. No signicant difference in the concentration of elements (Ca, Mn, Fe, Cu
and Zn) in the tissues of juveniles and adults of respective sex is observed, which indicated the active regulation of these elements by S. serrata.
2008 Elsevier Ltd. All rights reserved.

1. Introduction
Level of contaminants in the estuarine environment is increasing as a consequence of anthropogenic activities and result in
degrading water quality reects the status of aquatic resources
(Vijayavel et al., 2006). Mud crab, Scylla serrata commands a unique status by virtue of its delicacy and greater demand for consumption in local as well as export markets. With great demand
for live export and increased price, the shery and aquaculture of
mud crabs have gained worldwide importance (Mohapatra,
2008). Though the mud crabs are marine dwellers, they immigrate
into brackish water systems during their post-larval stages, grow
fast, attain maturity and form a lucrative shery in estuaries, backwaters and lagoons. Thus exposure to both marine and estuarine
environment has all effects on the body burden of metal bioavailability from the natural as well as anthropogenic set up of the
surroundings.
The interest on the accurate and easy analysis of trace elements
from the marine crustacean tissues arises from three areas of concern i.e. nutritional, toxicological and environmental. Some ele* Corresponding author. Tel.: +91 9861217792.
E-mail address: anil2k7@gmail.com (A. Mohapatra).
1
Present address: Coastal Ocean Monitoring and Prediction System Cell (COMAPS),
Institute of Minerals and Materials Technology, Bhubaneswar 751013, Orissa, India.
0278-6915/$ - see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.fct.2008.10.016

ments such as Mg, Al, Ca, Fe, Co, Cu and Zn are necessary for
maintenance of optimum health thus are important from nutrition
point of view in trace amount, metals such as Pb, Cd, As and Hg are
detrimental to optimum health and have toxicological effect and
the tissue samples are also used as the bio-indicator to assess bioavailability of contaminant concentrations in coastal water in environmental studies (Mohapatra et al., 2007). On the other hand
seafood is a signicant pathway to metal exposure in the coastal
areas (Hasmi et al., 2001) for which accurate and easy analysis of
concentration of trace elements from sea food is essential
(Mohapatra et al., 2007).
Various studies have already demonstrated the trace elemental
analysis from soft tissues for both biomedical and bioenvironmental samples by PIXE (Hota et al., 2001; Naga Raju et al., 2005;
Rautray et al., 2007). Analyzing the trace elemental concentration
from the sh otoliths (hard tissue) has been demonstrated by
external PIXE (Arai et al., 1996; Elfman et al., 1999). In the present
study we tried to demonstrate the effectiveness of the external
PIXE for analyzing both, soft tissues (body tissues for food quality
control purpose and environmental safety) as well as the hard tissues (Carapace for bioaccumulation and environmental safety
purposes).
The trace elemental analysis by external PIXE has lots of
advantages from biological point of view. The technique is fast,
sensitive, simultaneous and multielemental as well as for the

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A. Mohapatra et al. / Food and Chemical Toxicology 47 (2009) 119123

routine analysis, very simple sample preparation is needed. On the

other hand the technique is non-destructive and from very less
quantity to huge quantities of samples can be analyzed easily by
this technique. This technique is better than vacuum PIXE in the
sense that the time consumption for vacuum creation is saved.

for bioavailability of these elements. The detail of type and sources of pollution
along with its volume of discharge to the estuarine system is presented in Table
1. All the mud crab samples for the present study were collected from river mouth
point as this point receives the entire river burden of pollution and passes it to the
near coastal area.
2.2. Sample collection

2. Materials and methods

2.1. Study area
The Mahanadi river system is the third largest in the peninsula of India and the
largest river in Orissa state, India. The basin extends over an area approximately
141,600 km2, with a total length of 851 km and a peak discharge of 44,740 m3 s1
(Konhauser et al., 1997; Panda et al., 2006). The river begins in the Baster Hills of
Madhya Pradesh, ows over different geological formations of Eastern Ghats and
adjacent areas and joins the Bay of Bengal after dividing into different branches
in the deltaic area. The main branches of the Mahanadi meet the Bay of Bengal at
Paradip (Fig. 1). Along its course, the Mahanadi receives efuents from different
industrial and urban centers such as Sambalpur, Bauda, Cuttack, Choudwar, Jagatpur and Paradip in Orissa (Radhakrishna, 2001). Also the near by shing jetty close
to the river mouth adds certain source of pollution to the near coastal area through
the river mouth. It also receives a large amount of agricultural run-off along its
course. Human inuences are pronounced at Sambalpur, Cuttack and Paradip,
where the proliferation of industries (fertilizer, paper, textile, distilleries, etc.) and
sewage discharge is prominent (Sundaray et al., 2006). The major source of metal
pollution in Mahandi river sediments, water and suspended sediments are been
studied by various workers (Chakrapani and Subramanian, 1990, 1993; Konhauser
et al., 1997). The distribution and fractionation in bottom sediments of Mahanadi
basin demonstrated the higher availability of Mn and Cu in exchangeable fraction
(Chakrapani and Subramanian, 1993; Konhauser et al., 1997) which poses high risk

The mud crab samples were collected from the Mahanadi estuary (Fig. 1) by the
help of the sherman without any articial bait. The mud crab samples, selected for
the analysis were grouped as juveniles and adults from both the sexes in order to
evaluate the elemental variations in both the sexes separately as well as the juvenile and adult groups as the adult group is the main targeted group for their high
market demand as seafood both locally as well as abroad. The detailed biometry
of both sexes of juveniles and adults were given in Table 2. All the samples selected
for the analysis were in the intermoult stage. The crab samples were thoroughly
washed in de-ionised water after sacricing. All the samples were collected in the
same day during pre-monsoon. The different parts were separated by using sterilized stainless steel scalpel and forceps in order to avoid contamination. The different body tissues (Muscles, gill and carapace) were separated for the analysis in
external PIXE. The muscle and gill samples were lyophilized at Regional Medical Research Center, Bhubaneswar and directly taken for the proton beam irradiation with
out any pretreatment. The carapace samples were sun dried and directly exposed in
external PIXE for the elemental analysis.
2.3. Instrumental
The proton beam of energy 3 MeV obtained from the 3 MV tandem type pelletron accelerators was collimated by a graphite collimator to a beam size of 3 mm
diameter and the beam was extracted into air using a Kapton foil at the exit point
of a vacuum scattering chamber. The scattering chamber has an inner diameter of

Fig. 1. Sampling location with anthropogenic set up of the study area.

TM

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A. Mohapatra et al. / Food and Chemical Toxicology 47 (2009) 119123

Table 1
Sources of pollutants to Mahanadi estuary (Panda et al., 2006; Sundaray et al., 2006).
Serial
number

Major sewage/industries

Product

Main pollutants

Approximate volume of
discharge

Cuttack township

Sewage

708.94  106 m3 year1

2
3
4

Tatagarh paper mills, Chaudwar, Cuttack

Orissa textile mills Chaudwar, Cuttack
Paradip township

Pulp and paper

Cotton textile
Sewage

pH, BOD, nutrients, trace metals (like Zn, Cu, etc.)

3264 m3 day1

Fishing jetty (Near Mahanadi estuary)

Fertilizer and diammonium

phosphate
Fertilizer and diammonium
phosphate
Sewage and oil

4085 m3 day1

IFFCO (Previously OSWAL fertilizer plant

at Paradip)
Paradip Phosphates Ltd., Paradip (PPL)

Nutrients, BOD, oil, heavy metals, pathogenic

bacteria
BOD, COD, pH, Heavy metals
pH, BOD, dye, heavy metals.
Nutrients, BOD, oil, heavy metals, pathogenic
bacteria
pH, BOD, nutrients, trace metals (like Zn, Cu, etc.)

Not estimated

East Coast Brewaries and Distillaries Ltd.,

Paradip

Nutrients, heavy metal, petroleum hydro carbon,

BOD, nutrients
BOD, SS, pH, mineral acids, sugars

Brewaries

Table 2
Biometry of the Scylla serrata taken for the analysis of trace element analysis.
S. serrata (Life stages)

Carapace width (CW) in mm

Body weight (BW) in g

Adult (#)
Adult ($)
Juvenile (#)
Juvenile ($)

132 14
127 21
57 9
54 7

521 31
332 28
39 11
36 6

Table 3
Observed concentrations (lg/g) of various elements in certied reference materials
(CRMs) by external PIXE set up.
Elements

K
Ca
Mn
Fe
Cu
Zn
Se
Br
Pb

Certied reference materials (CRMs) bovine liver (NIST-1577b)

Certied value

Measured value

Percentage of recovery

9940 20
116 4
10.5 1.7
184 15
160 8
127 16
0.73 0.06
9.7
0.129 0.004

9916 467
115 13
10.3 1.2
191 18
157 10
130 11
0.71 0.05
10.1 1.1
0.128 0.003

99.76
99.14
98.10
103.80
98.13
102.36
97.26
104.12
99.22

70 m3 day1
1507 m3 day-1

376 m3 day1

absorber placed between detector and specimen. The instrumental constant H is the
product of the geometric solid angle of the X-ray detector and any systematic normalization factor present in the charge integration system. The instrumental constant H was measured using 3 MeV protons and a wide range of pure singleelement standards emitting both K and L X-rays in the energy region 326 keV. Thus
the X-ray yield (Y) is converted to elemental concentration via a dened standardization technique involving H value, the theoretical yield, detection efciency and lter transmission values. The error associated in the present study with the elements
is 812%.
2.4. Technique evaluation
For the checking of the reliability of the technique we had measured the certied reference material NIST Bovine liver (1577b) international standard and comparison of the certied value with the measured value is presented in Table 3.
The measured values and the certied values are in good agreement and thus the
experimental procedure adopted is reliable in analyzing the tissue samples.

3. Results and discussion

80 cm and was designed to cater to the requirements of the external beam. The
beam is rst focussed and centred at the target location inside the scattering chamber and then let through the thin Kapton foil placed at the exit port. The chamber is
pumped by a high throughput diffstack pump to maintain a vacuum in the range of
107 mbar in the chamber and the beam line. The Kapton foil is used as exit window due to its several special characteristics like low beam-induced background
emission, minimal energy loss and resistance to radiation damage. The beam is allowed to travel 3 cm in-air by which time the energy gets reduced to about 2.4 MeV,
and the proton beam then irradiates the samples. Beam charge measurement was
carried out using a rotating vane chopper of adjustable length (Rautray, 2008).
For irradiation with proton beam, the samples were kept in-air over a sample
stand making an angle 45 to the beam direction. The samples were irradiated with
maximum beam current of 15 nA passing through the 8 lm thick Kapton window.
A Si(Li) detector having energy resolution of 170 eV at 5.9 keV placed at 90 with
respect to the beam direction was used to detect characteristic X-rays emitted from
the sample. The detector has an active area of 30 mm2 and an entrance beryllium
window of 8 lm thickness. A 25 lm thick aluminised Mylar absorber (with 6% hole)
was kept in front of the detector to attenuate the bremsstrahlung background and
the dominant low energy X-ray peaks. Spectra were recorded by using a multi channel analyser, which was calibrated with 241Am X-ray source.
The data were analyzed with the GUPIX-2000 program. The relationship between characteristic X-ray yield Y (Z) for an element of atomic number Z and its
concentration Cz in a given target matrix is
TM

TM

Yz HY 1 zC z Q z t z
where Y1(Z) is the computed yield from the database per steradian per unit concentration and per unit integrated beam charge; Q the beam charge; ez the intrinsic efciency of the Si(Li) X-ray detector; and tz the transmission of the X-rays through any

The concentration of nine elements in the different tissue samples of S. serrata (muscle, gill and carapace) is presented in Table 4.
During the present study the concentration of nine elements (K, Ca,
Mn, Fe, Cu, Zn, Se, Br and Pb) were taken into consideration.
The concentration of K is higher in the body tissues of all the
specimens as compared to the other elements taken into consideration. The K concentrations in both the sexes were higher in the
muscle tissues of juveniles than the adults. Such observation was
also made from both S. serrata and S. tranquebarica from Chilika lagoon (Mohapatra, 2008). The K content in both the sexes were
higher in the gills than muscles and carapace in the adults whereas
in juveniles the K concentration was recorded higher in the muscles than other parts. In the present study the female crabs were
having less K concentration than the males in both adults and juveniles. The K concentration reported from the muscle tissues during
the present study is close to the observed concentration from S.
serrata and S. tranquebarica tissues from Chilika lagoon (Mohapatra
et al., 2007).
The Ca concentration was very high in the carapace than other
tissues which is obvious. Sex based difference in the Ca concentration was there i.e. the males having higher Ca concentration in all
the tissues than that of the females. Higher Ca concentration from
the male tissues of S. serrata and S. tranquebarica from Chilika lagoon was also reported (Mohapatra et al., 2007). The Ca concentration observed in the present study was slightly higher than the
same species from Chilika lagoon (Mohapatra et al., 2007) and lower in comparison to the reported value from the fresh water crab
Somaniathelphusa sinensis from Vietnam (Brauer et al., 2001).
In general, Mn concentrations are highest in calcied tissues
and lowest in edible esh of crustaceans (Eiser, 1981). It was

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A. Mohapatra et al. / Food and Chemical Toxicology 47 (2009) 119123

Table 4
Elemental concentration (lg/g) in body tissues of Scylla serrata in adult and juvenile stages of both the sexes.

Adult (#) n = 12 (M)

Adult (#) n = 12 (C)
Adult (#) n = 12 (G)
Adult ($) n = 12 (M)
Adult ($) n = 12 (C)
Adult ($) n = 12 (G)
Juvenile (#) n = 12 (M)
Juvenile (#) n = 12 (C)
Juvenile (#) n = 12 (G)
Juvenile ($) n = 12 (M)
Juvenile ($) n = 12 (C)
Juvenile ($) n = 12 (G)

Ca

Mn

Fe

Cu

Zn

Se

Br

Pb

9249.8 397
8156.6 296
9143.2 412
8804.2 416
8244.9 364
8956.6 466
9554.1 511
8037.4 368
8732.3 427
9023.9 518
8249.4 442
8828.5 417

3194.6 256
13567.5 569
3635.2 199
2598.4 229
12603.5 621
3215.4 302
3064.5 216
13070.6 620
3945 2.98
2586.2 251
12708.1 410
3858.7 346

12.8 1.3
18.6 2.0
18.5 1.7
11.2 1.3
24.8 2.1
17.2 0.9
13 0.6
19.7 1.3
18.6 1.9
13.8 1.2
26.1 2.3
14.2 0.9

159.7 13.2
125.9 10.6
190.4 16.2
171.4 12.9
112.6 9.7
182.8 18.1
163 6.9
117.8 9.6
194.3 12.9
175.2 15.3
119.1 11.2
181.8 19.1

114 10.4
47.9 3.2
32.9 2.9
132.3 13.0
51.6 5.2
31.6 2.4
109.5 10.1
43.9 3.7
29.1 1.8
113.1 8.9
42.6 4.6
26.1 1.9

279.3 19.6
63.4 4.9
42.2 2.9
312.6 28.6
71.6 6.9
52.8 5.9
261.7 29.3
65.4 6.9
47 3.9
296.8 26.7
73.7 8.1
56.7 5.9

0.47 0.030
0.33 0.011
0.43 0.021
0.39 0.006
0.29 0.007
0.41 0.009
0.42 0.012
0.32 0.009
0.39 0.006
0.44 0.009
0.34 0.011
0.39 0.018

189.3 11.3
23.9 3.2
46.5 3.9
163.8 14.9
30.6 2.8
41.8 3.6
211 18.6
30.8 3.1
48.6 4.6
243.4 24.3
37.1 2.9
42.7 4.1

0.19 0.006
0.16 0.01
0.18 0.009
0.23 0.013
0.18 0.008
0.13 0.010
0.26 0.009
0.15 0.003
0.160 0.06
0.2 0.007
0.14 0.011
0.14 0.009

M: muscle tissues; C: carapace; G: gill; n: number of samples.

conrmed as the carapace contained higher levels of Mn than the

gill and body tissues in both juveniles and adults irrespective of
sexes. The Mn concentration was observed higher in females than
that of the male muscle tissues. Such observations were also made
from the S. serrata and S. tranquebarica from Chilika lagoon (Mohapatra et al., 2007; Mohapatra, 2008) and from crustaceans like Panulirus inatus from pacic coasts (Paez-Osuna and Ruiz-Fernandez,
1995a). Not much difference in the Mn concentration in juvenile
and adult muscle was observed.
Iron plays a vital role in the enzymatic and respiratory processes of crustaceans (Frenet and Alliot, 1985; Paez-Osuna and
Ruiz-Fernandez, 1995a). In the present study the Fe content was
recorded higher in the female muscle tissues than the males irrespective of life stages. The higher Fe concentrations in the female
tissues were also reported from the tissues of other crustacean
samples from the globe by various workers (Paez-Osuna and
Ruiz-Fernandez, 1995b; Ruelas-Inzunza and Paez-Osuna, 2004;
Mohapatra et al., 2007; Mohapatra, 2008). The Fe concentration
was higher in the gills than that of the carapace and muscle tissues.
The higher Fe content in gills of the mud crabs may be due to the
presence of absorbed particulate matter on gills rather than to active biological uptake of the metal (Szefer et al., 1990; Paez-Osuna
and Ruiz-Fernandez, 1995a). Similar high values of Fe from gills
has been reported previously for Cancer irroratus (Martin, 1974),
S. serrata (Depledge et al., 1986), P. inatus (Paez-Osuna and
Ruiz-Fernandez, 1995a) and P. vannamei (Paez-Osuna and Tronmayen, 1996).
The Cu concentrations in the muscles were always higher than
the gill and carapace samples. The Cu concentration in the present study is close to the values recorded from Portunus pelagicus
from the Kuwait coast (Al-Mohanna and Subrahmanyam, 2001).
The higher concentration of Cu from the muscle than the gills
from crab (P. pelagicus) was also reported by Al-Mohanna and
Subrahmanyam (2001). The Cu concentration was higher in the
female muscle tissues than the males; such observations were
also made by Mohapatra et al. (2007) from the muscle tissues
of S. serrata and S. tranquebarica from Chilika lagoon. The adult
mud crabs were having slightly higher concentration of Cu than
the juveniles. Though Cu is an essential element in marine organisms, is thought to be strictly regulated in muscle tissues (Phillips,
1980; Al-Mohanna and Subrahmanyam, 2001). The higher concentration of Cu in the muscle tissues in comparison to other
studies might be due to the impact of near by fertilizer plants
and industrial activities. Such source of Cu pollution by fertilizer
plants are also been reported previously (Al-Mohanna and
Subrahmanyam, 2001). On the other hand, Chakrapani and Subramanian (1993) reported higher enrichment of Cu in exchangeable fraction in Mahanadi river sediments due to strong afnity
of organic waste with Cu. The bioavailability of Cu in the Maha-

nadi sediment may be responsible for bioaccumulation in tissues

of the mud crabs.
In the present study the Zn concentration is always higher in
the body tissues followed by the carapace and gill, irrespective of
sex and size. The Zn concentration in the present study is close
to the same species from Chilika lagoon (Mohapatra et al., 2007).
The higher concentration of Zn in the muscle tissues of other decapods from Mexico were also reported by Paez-Osuna and Ruiz-Fernandez (1995a). The Zn concentration was recorded higher in the
female tissues than the male tissues (Paez-Osuna and Ruiz-Fernandez, 1995a; Ruelas-Inzunza and Paez-Osuna, 2004; Mohapatra
et al., 2007; Mohapatra, 2008). No signicant variation in the trace
elemental concentrations of the juveniles and adults was observed
though slightly higher concentration of Zn was recorded from the
adult tissues which suggest that the S. serrata might be capable
of regulating the Zn concentration in their body tissues. There is
sufcient evidence that the decapods are capable of regulation of
Zn in their body (Rainbow, 1985; Bryan, 1976; Paez-Osuna and
Ruiz-Fernandez, 1995a). But higher concentration of Zn was also
reported for other marine invertebrates from relatively polluted
areas (Halcrow et al., 1973; Rainbow, 1988; Brauer et al., 2001)
which suggest that the regulation of this element may not be
complete.
The Se concentration in the mud crab tissues was very low in
comparison to the safe limits i.e. <24 lg/g (Hamilton, 2004) and
Pb concentration was also observed to be very low and well with
in the permissible limits (1.5 lg/g) for seafood (CAA, 2006) from
human consumption point of view. The Pb concentration in the
present study is very low in comparison to the other studies from
decapods (Bryan, 1976; Anderlini et al., 1982; Al-Mohanna and
Subrahmanyam, 2001; Hossain and Khan, 2001) and close to the
study by Mohapatra et al. (2007) for the same genus (S. serrata
and S. tranquebarica) from Chilika lagoon. The concentration of
Br, from the body tissues of the mud crabs in the present study closely in the range reported by Mohapatra et al. (2007). The concentration of Br is always higher in the juveniles than that of the
adults. The concentration of Br is in the order of muscle > gill > carapace in the present study. The concentration of Br in the muscle
tissues are very close to the observed values from the muscle tissues of S. serrata from Chilika lagoon (Mohapatra et al., 2007).
The distribution of different elements in the tissues samples of
crustaceans reported previously from different locations were presented in Table 5. The variations in the concentration of elements
crustacean tissues of different geographical locations might be
due to the difference in the ecological and anthropogenic set up
of that particular area. In the present study there observed a clear
sex based difference in the trace elemental concentrations. Some
elements like K and Ca were higher in the male tissues. Some
elements (Fe, Cu, Zn and Pb) in the body tissues of females were

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A. Mohapatra et al. / Food and Chemical Toxicology 47 (2009) 119123

Table 5
Concentration of various elements in muscle tissues samples of crustaceans from different geographical locations in lg/g.
Geographical area

Species

Reference

Vietnam
Kuwait
Kuwait
World wide
Bay of Bengal
Chilika lagoon, India
Mahandi estuary, India

Somaniathelphusa sinensis
From crustacean samples
Portunus pelagicus
From 24 references
P. monodon
Scyll sp.
Scylla serrata

Brauer et al. (2001)

13700
Anderlini et al. (1982)

Al-Mohanna and Subrahmanyam (2001)

Bryan (1976)

Hossain and Khan (2001)

Mohapatra et al. (2007)

9191
Present study
9158

higher than the males, which may be due to the slow growth rate
(Mohapatra, 2008) of the female crabs than the male thus higher
accumulation of these trace elements. The faster growing sex can
be expected to contain lower concentration of metal (Pourang
et al., 2004). But a very little difference in the trace elemental concentrations (Ca, Mn, Fe, Cu and Zn) in the muscle tissues of juveniles and adults of respective sex gives the indication that the
crustaceans regulate the body burden of trace elements from the
environment by their metabolism.
The study clearly demonstrated the effectiveness of the external
PIXE in analyzing the elemental concentrations from the tissue
samples for environmental study and opens the door to carry out
the accurate and effective analysis of both soft and hard tissue
by a non-destructive, simultaneous, multi-elemental technique
with a very little sample preparation.
Conict of interest statement
The authors declare that there are no conicts of interest.
Acknowledgements
Author (A.M.) is grateful to Mr. S.K. Mohanty, Ex-Joint Director,
Department Fisheries, Orissa, for his most valuable suggestions.
Authors sincere thanks to Mr. M.K. Khadanga and Mr. P. Samantaray, Research Scholars of Institute of Minerals and Materials Technology, for their help in sample collection. We are also thankful to
Girija and Kuku for their help in lyophilizing the samples.
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Ca

Mn

Fe

Cu

Zn

Se

Br

Pb

15200

2404
2861

28
0.98
0.95
1.0
3.16.5
12
12.7

56

9.115.7
161
167.3

62
14
123.8
70
12.121.2
124
117.2

420
46
206
80
24.135.7
280
287.6

1.3

0.38
0.43

40

223
201.9

0.7
1.9
1.0
0.81.2
0.18
0.22

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