Pakistan Journal of Scientific and Industrial Research · Web view1973; Khan, 1992; Adeyeye, 2002; John-Samuel et al., 2004). Portunus sanguinolentus is one of the important representatives

Elemental distribution analysis of three-spot swimming crab shell

Dear Editor, PJSIR Journal Karachi,Subject: process of Article code 10297-ZA/ 3063Respected Sir/Madam,Many more thanks for consideration of request for publication of this article code 10297-ZA/ 3063. The article entitled below is submitting for evaluation and publication process

Elemental Distribution Analysis of the shell of Three-Spot Swimming Crab Portunus sanguinolentus (Herbst, 1783) from the coastline of

Karachi, Pakistan1Wajeeha Razzaq, 1Zubia Masood*, 2Semra Benzer and 3Quratulan Ahmed

1. Department of Zoology Sardar Bahadur Khan Women’s University, Quetta, Balochistan

2. Gazi University, Education Faculty, Science Education, Teknikokullar Ankara 065003. Marine Reference Collection and Resource Centre, University of Karachi

*Corresponding Author: Dr Zubia Masood Permanent Address House No. 121-R,Sector G, Bhatai Colony, Korangi Crossing, Karachi Email: [email protected]: 03454881576

No of pages =14No of Tables=2Number of Figures=6The names of evaluators/referees are as follows;

1. Dr. Paolo Merella Parassitologia e Malattie Parassitarie Dipartimento di Medicina Veterinaria Universita' di Sassari via Vienna, 2 07100 Sassari Tel +39 079 229456 Cell +39 328 0334217 Fax +39 079 229464 e-mail [email protected]

2. Dr. KATSELIS GeorgeProfessorHead of Department of Fisheries and Aquaculture TechnologyTechnological Education Instituteof Western Greece (TEI/WG), TK 30200GREECE,Email id: GKatselis <[email protected] 2631058237,202

3. Prof. Dr. Muhammad ShafiDean Faculty of Marine Sciences,Lasbela University of Agriculture, Water and Marine Sciences, Uthal, Balochistan, Pakistan, Email: [email protected]: 0334-3554013

4. Professor Dr Wazir Ali BalochDepartment of freshwater Biology and Fisheries,University of Sindh, Jamshoro, PakistanEmail: [email protected]

1

mailto:[email protected]




Elemental Distribution Analysis of the shell of Three-Spot Swimming Crab Portunus sanguinolentus (Herbst, 1783) from the coastline of

Karachi, Pakistan

ABSTRACT

A study was conducted for the detection of elements in the shells of three-spot swimming

crab, Portunus sanguinolentus collected from the Karachi coast during the months of January

to March 2017. A total of 30 crab shell samples were collected and divided into three

replicates (10 shells for each samples). The element detection was performed by Energy

Dispersive X-ray spectroscopy with Scanning Electron Microscope (SIM/EDAX). The mean

concentration of elements includes i.e., carbon (C), oxygen (O), calcium (Ca), copper (Cu),

magnesium (Mg) and phosphorus (P) observed in the shell of P. sanguinolentus was

13.6±6.17%, 48.3±14.3%, 34.7±18.9%, 3.18±1.24%, 1.14±0.99% and 1.72±1.08 %,

respectively. The distribution pattern of these observed elements in shells of P.

sanguinolentus was found in the following order: O>Ca>C>Cu>P>Mg, respectively. From

the obtained results, it was concluded that crab shell can act as a good biosorbent for several

kinds of minerals existing in its ecosystem. Thus, it was concluded that crab shell is a good

indicator of certain metals found in its environments. Our present findings proved that crab

shell is a rich source of certain minerals that can also be utilized in extraction and preparation

of several pharmaceuticals and various other products in the future.

Key words: Portunus sanguinolentus, Crab shell, Elemental distribution.

Introduction

Crab is typically a short-tailed decapod crustacean that occurs all over the world,

particularly in tropical and semi-tropical regions. It can lives in oceans, freshwater

ecosystems, as well as on lands. About 850 crab species had been reported throughout the

world. Crabs are mostly omnivores that feed primarily on algae, mollusks, other small

crustaceans, worms, bacteria, fungi and detritus, but also depend on the availability of food as

well as on the type of species. Crab is also serves as a source of seafood in many parts of the

world, particularly in Japan, European and North American countries. Few species are eaten

as whole (usually soft-shell crabs), while in case of hard-shell crab, just their claws or legs

are mostly used for human consumption (Zafar and Ahsan, 2006; Sudhakar et al., 2009).

Crab can also be considered as an indicator of aquatic pollution of its habitat because of the

2


biosorbent ability of crab shells (An et al., 2001). It’s a rich source of protein and also have

great medicinal, socioeconomic and cultural values, therefore, more attention are required to

explore crab fishery for fulfilling their increasing demand (Hossain et al., 2015; Varada et al.,

2016). More recently, Krisfalusi-Gannon et al. (2018) had observed the increasing its

commercial demands of horseshoe crab in biomedical industries.

Nowadays, crab industry is being fast developed in the whole world; as it is highly

abundant shell-fish that constitute 20% of all crustaceans, which are caught as well as farmed

throughout the world. About 1.4 million tons of different crab species including i.e., horse

crab (Portunus trituberculatus) that accounts one quarter of it. Whereas, about 98% this total

catch of horse crab is being caught from the China coast. In addition, the other most

important crab species, such as, flower crabs (Portunus pelagicus), blue crabs (Callinectes

sapidus), edible crabs (Cancer pagurus), snow crabs (Chionoecetes), mud crabs (Scylla

serrata), and dungeness crab (Metacarcinus magister), which also served as economically

important food resources for many years in international and national open markets (Hassan

and Zafar, 2013; Hossain et al., 2018). In 1993, the harvest of blue crabs was valued at

around 100 million U.S. dollars (Sudhakar et al., 2009). Hagashi et al. (1979) and Kathirvel

(1993) had reported that due to very delicious taste and large size, the demand of live mud

crabs, Scylla serrata have now been increasing, which also raise its price in both national and

international markets. Moreover, both swimming crab species like Portunus sanguinolentus

and Portunus pelagicus now are being exported mostly in frozen or canned forms. As natural

availability of many other crab species is limited in few seasons only, but the abundance of

both swimming crab species, Portunus sanguinolentus and Portunus pelagicus remain same

throughout the year. The swimming crab species (Portunus sanguinolentus and Portunus

pelagicus) are major fishery resources of the seas of South East Asia. These species have now

elevated value from commercial perspective in various countries includes i.e., Bangladesh,

India, Arab countries and Southeast Asian countries. These species are very importance for

medical point of view because of their used during pregnancy by women. They contain high

levels of amino acids that help to control various inflammatory, rheumatoid arthritis, Crohn’s,

and cardiovascular diseases (Rao et al., 1973; Khan, 1992; Adeyeye, 2002; John-Samuel et

al., 2004).

Portunus sanguinolentus is one of the important representatives of decapod

crustacean and commonly found in the Karachi coasts of Pakistan. It is carnivore and found

in sandy and muddy coast. It is commercially harvested edible crab species in many parts of

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the world and also possesses great capability for accumulating certain heavy metals. As

certain elements can play a significant role in ecotoxicology, because of their highly

persistent and found to be toxic particularly for benthic organisms. The shell of this species

have also been used for detecting its ability to accumulate metals from water more than its

other soft body tissues as previously reported by An et al. (2001), Viswanathan et al. (2013),

Sarkar et al. (2016), Bat et al. (2018) and Saher & Kanwal (2018), respectively.

Crab fishery plays an important issue in the national economy of Pakistan. After

shrimps in Crustaceans, crabs were reported as 25,000 biomass, maximum sustainable yield

8,500 and production 4,218 tons for 2006 (Kanwal and Saher, 2016). In order to get

information of background amounts of elements and their fluctuations in bioindicator

organisms is essential as well as a thorough understanding of accumulation and detoxification

strategies. In this study, therefore the amounts of some elements in the shells of P.

sanguinolentus from Karachi shores of Pakistan were determined. As shells of crab are also

caught along with other invertebrates occurs in shallow sea shore, but they have very less

marketing demand, and therefore usually discarded from the landing sites. Therefore, this

study would be valuable in observing the medical and economic value of this crab species

and also stop the wastages or discarding of inconsumable part i.e., shell of crabs in order to

increase its suitable used in biomedical industries and also prevents ecological toxicity.

Materials and Methods

1. Study Area

A total of 30 crab samples of Portunus sanguinolentus samples were collected from

the Clifton Beach (Sea view) of Karachi coast located at 24.78 °N latitude and 67.04°E

longitudes on North-eastern border of Arabian Sea during the period from January to March

2017 (Figure 1). It is sandy coastline and also opens for all visitors from local population.

Therefore, this area was reflecting the progression of any contamination, ecological damages

and various other destructive activities of human here.

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Figure 1. Google Map of Clifton Beach of Karachi coast.

2. Sample preparation

Crab samples were immediately transfer in plastic bags and transferred to the

laboratory in the department of Zoology, University of Karachi for further analysis. After

sampling, each collected specimen was identified by using field guide of Bianchi (1985), as

shown in Figure 2, respectively. Then these samples were rinsed carefully using sterile

distilled water. Whole soft tissues were then separated carefully from the shells. These air

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Figure 2. Shell of P. sanguinolentus Figure 3.Crab shell Powder

Figure 4. Grinding of shell of P. sanguinolentus in mortar to powdered form


dried hard shell samples were then divided in to three replicates (10 shells for each sample).

After credentials, each replicate was instantly kept on digital balance and its dry-weight was

measured in grams before grinding into very fine powdered form by using mortar (Figure 4).

After grinding, 10 g of powdered shells obtained from each replicate was stored in cleaned

polythene bags and were then sent to Central Science Laboratory of University of Karachi for

elemental detection by EDAX (JED-2300 Analysis Station) with SEM (JSM-6510LA. The

system is capable of qualitative analysis, quantitative analysis, and elemental map on a thin-

film specimen with high energy resolution and high sensitivity. For SEM studies, one gram of

powdered shell obtained from each replicate were mounted on metallic stubs by using double

adhesive tape and coated with 100°A thick gold layer in gold coating unit for overcome the

issues of beam damage. Images were viewed under vacuum at an accelerating voltage of 20

Kv in SEM/EDAX and recorded on photographic plate (Figure 5). Specifications of SEM

images of three replicates A, B & C were given in Table 1, respectively. After SEM, shell

powder replicate samples A, B & C were used for EDAX study separately, and then element

distribution was analyzed. All process of sample collection, preservation and preparation in

the laboratory were made according to methods as followed by Bernhard (1976) and Thakur

et al. (2016). Data obtained in this study was subjected to descriptive statistical analysis

includes range, means, and standard deviations. All these analyses were made by using SPSS

v.11.5 statistical software.

Table 1. Specifications of SEM images obtained for three replicates A, B and C was mentioned below as follows;

Acquisition Parameter Sample A Sample B Sample CInstrument 6380(LA) 6380(LA) 6380(LA)Acc. Voltage 20.0 kV 20.0 kV 20.0 kVProbe Current 1.00000 nA 1.00000 nA 1.00000 nAPHA mode T3 T3 T3Real Time 38.91 sec 40.85 sec 44.03 secLive Time 30.00 sec 30.00 sec 30.00 secDead Time 22 % 25 % 32 %Counting Rate 4720 cps 5439 cps 7198 cpsEnergy Range 0 - 20 keV 0 - 20 keV 0 - 20 keVVolt 20.00 kV 20.00 kV 20.00 kVMag x 5,000 x 8,000 x 10,000Pixel 1280 x 960 1280 x 960 1280 x 960

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Results and discussion

In this study, three replicates were made in order to analyze with good accuracy as

followed the method of Thakur et al. (2016). The concentration of each element of the

present study was expressed as percentage of element per gram (g) of shell powder, and the

obtained results of SEM/EDAX for three replicates A, B & C were presented in Table 2 and

Figure 6, respectively. In the present study, eight elements were observed in the hard shell of

P. sanguinolentus, respectively. The mean concentration of these elements includes i.e.,

carbon (C), oxygen (O), calcium (Ca), copper (Cu), magnesium (Mg) and phosphorus (P)

observed in the shell of this crab species was 13.6±6.17%, 48.3±14.3%, 34.7±18.9%,

3.18±1.24%, 1.14±0.99% and 1.72±1.08 %, respectively. Whereas, the distribution pattern of

all these observed elements was found in the following order: O>Ca>C>Cu>P>Mg,

respectively. Our present results are very much comparable with Sudhakar et al. (2009), who

also reported six elements like calcium, magnesium, sodium, potassium and zinc in the shell

of Portunus sanguinolentus found along Indian coast, and among them, calcium was found in

highest concentration of calcium than other elements found in crab shell, which was also in

agreement with our present study. Likewise, María et al. (2011) had also found the higher

concentration of calcium than phosphorus and magnesium in the shells of snow crab

(Chionoecetes opilio). Ferraro et al. (2010) and Lee et al. (2010) also reported that crab shell

can also serve as as rich source of minerals, particularly calcium in food sectors, and have

been successfully utilized as a major feed ingredient for cod and rainbow trout fishes. While

in contrast, both sodium and potassium had form the major portion of mineral composition in

the shell of boiled crab as reported by Hagashi et al. (1979). According to Chen et al. (2008),

minerals of crab shell was found in the form of amorphous calcium carbonate deposited along

with chitin-protein matrix. Furthermore, the concentration of calcium can be used for

observing the hardness of crab shell; however, such amount was found to be varies according

to type of species, as well as within one shell. Kanwal and Sehar (2016) have been observed

that such variations in the biochemical composition might be varies because of season,

temperature, body size, maturity stages, accessibility of food, aquatic pollution and different

geographical location of habitats. Most marine animals are mostly rich source of certain

minerals. Both fish and shellfish can absorb certain minerals from their environment with the

help of their gills, skin or other body surfaces. Almost all elements are found in seawater and

also to some extent in its aquatic animals. Among them, the most abundant elements of

seawater including i.e., Sodium (Na), potassium (K),calcium (Ca), phosphorus (P),

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aluminium (Al), Barium (Ba), Cadmium (Cd),iodine (I),Copper (Cr), lead (Pb), lithium (Li),

mercury (Hg), silver (Ag) and vanadium (V). Gopakumar (1997) reported that ash content in

fish was found in ranged from 0.5 to 2.0%. As minerals are serve as major components in

bones or soft tissues and also act as co-factors or co-activators during various important

enzymatic reactions in human nutrition. Calcium, phosphorus, magnesium, sodium and

potassium have been considered as macro elements that are required for normal body

functions and involved in the maintenance of membrane potential. Both calcium and

phosphorus constitutes 70 to 80% portion of exoskeleton in fishes as reported by Nair and

Mathew (2000).

Conclusions

From the present results, it was concluded that the knowledge of elements occurs in crab shell

is highly important for regulate the quality of natural ecosystem as well as its management, and helps

in determine the contaminated places. Therefore, our present information regarding to the the

distribution pattern of elements could be the indication of their presence in marine environment and

final transfer to human with the help of seafood consumption.

Acknowledgements

We expressed my gratitude to Professor Dr. Rehana Yasmeen and Musarrat-Ul-Ainfrom

department of Zoology of Karachi University for their kind support in samples preparation in labs and

also help for conducting the metal analysis in Central Science Laboratory of Karachi University. Also

special thanks to my research supervisor, Dr. Zubia Masood for her help and guidance in the

preparation of this manuscript.

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1

Table 2. Concentration of elements measured in mass% per gram of crab shell powder

Sample A Sample B Sample C

Elements Mass% Atomic%

Mass% Atomic%

Mass% Atomic%

Mean± SD

Carbon (C) 8.70 16.20 11.6 18.20 20.6 28.20 13.6±6.17

Oxygen(O) 32.0 46.20 53.9 60.20 58.86 61.01 α48.3±14.3

Calcium(Ca) 55.6 33.20 29.7 19.00 18.86 7.90 34.7±18.9

Copper (Cu) 3.55 1.30 4.21 0.41 1.80 0.10 3.18±1.24

Magnesium(Mg) 0.01 1.32 1.75 1.10 1.68 1.11 *1.14±0.99

Phosphorus (P) 1.14 1.80 2.98 1.10 1.06 1.70 1.72±1.08

*Note: α shows the highest mean metal concentration, * shows the lowest mean metal concentration


C O Ca Cu Mg P0

10

20

30

40

50

60

Figure 6. percentage composition of element in shell of P. sanguinolentus

ABC

2


Figure 5. Scanning Electron Microscope (SIM) Images of Sample A, B & C of Crab shell powder.

1

A B

C

Documents

Pakistan Journal of Scientific and Industrial Research · Web view1973; Khan, 1992; Adeyeye, 2002; John-Samuel et al., 2004). Portunus sanguinolentus is one of the important representatives