Biosorption of metals by Solitary Ascidian, Phallusia Nigra in Vizhinjam

Bay (South West Coast of India)

Keywords:
Ascidian, biosorption, metals, Phallusia nigra, Vizhinjam Bay.
060-067 | JRAS | 2013 | Vol 2 | No 1
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Journal of Research in
Animal Sciences
An International Scientific
Research Journal
Authors:
Abdul Jaffar Ali H
1
*,
Tamil Selvi M
2
and
Bakavathiappan GA
3
.




Institution:
1.Department of
Biotechnology, Islamiah
College, Vaniyambadi -
635752, Tamilnadu, India.

2. Department of Zoology,
V.V. Vanniyaperumal
College for women
Virudhunagar- 626001,
Tamilnadu, India.

3. Department of Zoology,
S.B.K. College,
Aruppukottai - 626101.



Corresponding author:
Abdul Jaffar Ali H.












Email:



Web Address:
http://janimalsciences.com/
documents/AS0022.pdf.
Dates:
Received: 17 Oct 2013 Accepted: 24 Oct 2013 Published: 28 Oct 2013
Article Citation:
Abdul Jaffar Ali H,Tamil Selvi M

and Bakavathiappan GA
Biosorption of metals by Solitary Ascidian, Phallusia Nigra in Vizhinjam Bay (South
West Coast of India)
Journal of Research in Animal Sciences (2013) 2(1): 060-067.
An International Scientific Research Journal
Original Research
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ABSTRACT:
Biosorption of heavy metals by ascidians has been recognized as a potential
alternative to existing technique for recovery of heavy metals from waste stream. The
solitary ascidian, Phallusia nigra, is a common and large black ascidian and occurs
throughout the year in Vizhinjam Bay. Hence this study has been focused on the
biosorption of metals in the test and mantle body of P. nigra from ecologically
significant station, Vizhinjam Bay. Monthly samplings of water and specimens of
P. nigra were done during the period from September 2010 to August 2011 and
subjected to the analysis of metals. The concentration of metals in water were found
in the following order Cd>Pb>V>Hg. Mean concentrations of metals were high during
monsoon season and low during premonsoon season.
Absorption of metals by P. nigra revealed that both test and mantle body
accumulated a diverse range of trace metals. The present results showed that the
content of metals in the test was usually less than that of the mantle body. High
concentration of these metals was found to be during monsoon season whereas low
concentration was during premonsoon season.
The bioaccumulation factors were in the order of V>Pb>Cd> Hg for the test
and mantle body of the study animal. Application of one way ANOVA for the
concentration of these metals between test and mantle body showed significant
differences. Metal concentrations recorded in this ascidian could effectively be used
as good reference material for monitoring metal contamination in Indian sea waters.


INTRODUCTION
The entry of many trace metals from terrestrial
and atmospheric sources to the marine environment has
increased considerably in recent past. These metallic
species that are released tend to persist forever and
accumulating in living tissues through the food chain
posing serious problems. In this scenario, studies about
removal / recovery of heavy metals from marine
environment are need of the hour.
Though there are plenty of methods available,
these processes may be ineffective or expensive
(Volesky, 1990). Therefore, the research for new cost
effective technologies for the removal of heavy metals
from the environment has been directed towards
biosorption. Bioremediation has emerged in the last
decade as one of the most promising alternatives for the
control of metal pollution. The use biological organisms
to abate metal pollution have advantages over
conventional treatment methods due to low cost, high
efficiency, ecofriendly, regeneration of biosorbents and
possibility of metal recovery (Kratochvil and Volesky,
1998).
Studies on better understanding of metal
biosorption by certain potential biosorbents are available.
These biosorbents include some fungi, algae, bacteria,
yeast and agricultural waste. Recently, biosorption of
heavy metals by ascidians has been recognized as a
potential alternative to existing technique for recovery of
heavy metals from industrial waste stream.
Henze (1911) was the first to show that ascidians
are capable of accumulating vanadium in their bodies as
a complex organic molecule. Monniot et al., (1993)
found that phelobobranchs principally store the metals in
all their tissues and were the best indicators for metallic
and organo-metallic pollutions. Though significant
literatures are available for the distribution of various
heavy metals in Indian seas, use of ascidians as sentinel
organisms in biosorption of heavy metals has been little
investigated. Preliminary works on accumulation of
metals in ascidians have also been studied by Krishnan
(1992) and Abdul Jaffar Ali (2004).
The characteristics of sedentary ascidians to
accumulate metals in excess of the environment make
them the interesting research objects for testing and
modeling the marine ecosystem. Therefore, it is
imperative to understand the potential functions of
ascidians as a biological filter of the aquatic
environment. The solitary ascidian, Phallusia nigra, has
been a suitable candidate for biomonitoring studies
because of its sedentary lifestyle, abundance and
continuous breeding, easy identification and sampling.
This black ascidian is abundant in Vizhinjam Bay.
In view of their potential to entrap several toxic
heavy metals and lack of literature on the metals
composition of ascidians in general from Indian coast,
the present study was aimed at understanding the
availability of the biosorption of metal ions such as,
vanadium, cadmium, lead and mercury by solitary
ascidian, Phallusia nigra from Vizhinjam Bay (south
west coast).

MATERIALS AND METHODS
For the present investigation, ecologically
significant station, namely Vizhinjam situated along the
south west coast of India was chosen.
Study animal
Phallusia nigra Savigny, 1816, has been
recorded abundantly from the Indian waters eg.
Vizhinjam (South West coast). This common large
solitary ascidian is typically velvety black or dark brown
in colour. This animal breeds throughout the year.
Description of the study area
Vizhinjam Bay, located in the geographical
coordinates of Long 765615E Lat 82230N, is
situated 16 km south to Trivandrum city. This station and
the nearby coastal areas may be vulnerable to
contamination with human faecal matters and sewage but
less due to industrial effluents. Vizhinjam coast is
Ali et al., 2013
061 Journal of Research in Animal Sciences (2013) 2(1): 060-067
subjected to pronounced seasonal changes in climatic
regions with distinct seasons like Premonsoon (February
May), Monsoon Season (June September) and Post
Monsoon (October January). The bulk of rainfall in
this area is due to south west monsoon.
Trace metal analysis
The monthly water samples were collected from
September 2010 to August 2011 in the habitat of the
species chosen at the study area in pre-cleaned and acid
washed polypropylene bottles. After filtration in
Millipore filter paper (mesh size 0.45 ), the resulting
solutions were analysed for metal analysis.
Known specimens of the solitary ascidian,
Phallusia nigra were also collected monthly. They were
washed and dissected to separate test (body covering)
and mantle body (soft bodied part) for the analysis of
these metals. The dissected portions were dried in an
oven at 110C for twenty four hours. A known quantity
of each material was powdered and acid digested in a
mixture of nitric acid and perchloric acid (2:1 v/v)
(F.A.O. 1983). The residue was dissolved in 10 ml of 2
N Hydrochloric acid. Both the water and specimen
samples for vanadium, cadmium and lead were analysed
in GBC-Aventa (Ver 1.33) Atomic Absorption
Spectrophotometer. Mercury was analysed by cold
vapour technique using mercury analyzer (OPMEC
CECRI, Port Trust, Tuticorin).
Concentrations of metals in test and mantle body of
P. nigra were presented as means SD and subjected to
one way ANOVA for testing the significance of the
difference in the concentration between the test and
mantle body.

RESULTS AND DISCUSSION
Metal level in sea water
Metals such as vanadium, cadmium, lead and
mercury occur in the seawater in different forms at
different concentrations. Annual mean concentrations of
metals in seawaters varied seasonally at Vizhinjam.
Seasonal variations in the concentration of metals in the
sea water from the study area are depicted in Figures 1.
In water, the average value of 0.0012 ppm was
found to be maximum during monsoon and minimum
during premonsoon. Mean cadmium level in sea water
was found to be 0.0016 ppm with maximum (0.003 ppm)
during monsoon. The concentration of lead was
maximum (0.003 ppm) during monsoon season and
minimum (0.001 ppm) during premonsoon season. Trace
level of lead was also observed during February and
March 2011. On an average, 0.9 ppb of mercury level
was recorded.
These results revealed the fact that the
concentrations of metals were high during monsoon and
minimum during premonsoon. This could be
Ali et al., 2013
Journal of Research in Animal Sciences (2013) 2(1): 060-067 062
Figures 1. Seasonal variations in the concentrations of metals in the sea water from Vizhinjam.


corroborated with the removal of a substantial portion of
these metals from water by phytoplankton and binding to
other suspended matters.
Biosorption by the solitary ascidian, P. nigra
The mean concentration of trace metals in
P. nigra from Vizhinjam is given in Table 1. Seasonal
variations in the accumulation of metals in the test and
mantle body of P. nigra are shown in the Figures 2-5.
Absorption of metals by P. nigra revealed that both test
and mantle body accumulated a diverse range of trace
metals. Their detection in ascidian indicates the
recalcitrant nature of these contaminants in the
environment. The present results showed that the content
of metals in the test was usually less than that of the
mantle body. It is generally opined that the metals are
concentrated in the body through food chain. Ascidian,
as a sedentary organism, filters a large volume of water
and feed large amount of deposit and plankton, which
accumulate heavy metals as suggested by Laws (1981),
who reported the high concentration metals in the
phytoplankton than in seawater. The concentration of
metals in zooplankton is substantially higher than the
phytoplankton concentration (Rejomon et al., 2008).
The concentration of Vanadium in the test and
the mantle body of P. nigra collected at the study area
ranged from 79.75 ppm to 92.05 ppm and 229.06 ppm to
335.23 ppm respectively (Fig 2). Maximum level of
concentration was observed during monsoon for both test
and mantle body whereas the minimum was during
March (test) and May, 2011 (mantle body)
(Premonsoon). Among the four metals, vanadium has
been found to highly elevated levels in the study animal
at both stations. Stephen et al., (2008) reasoned the high
level of vanadium in the ascidian blood to involvement
Ali et al., 2013
063 Journal of Research in Animal Sciences (2013) 2(1): 060-067
STATIONS
Vanadium (ppm) Cadmium (ppm) Lead (ppm) Mercury (ppb)
Test Mantle body Test Mantle body Test Mantle body Test Mantle body
VIZHINJAM
Mean 85.54 274.10 0.980 5.420 12.75 20.78 18.814 28.262
S.D 3.71 33.18 0.577 4.079 7.14 13.50 1.211 5.770
Table 1. Trace metal level in Phallusia nigra between two stations. Each value is the mean of
3 estimations with standard deviation (S.D).
Figures 2. Seasonal variations in the accumulation of vanadium in the test and mantle body of
Phallusia nigra at Vizhinjam Bay.
of tyrosine derived tunichrome tripeptides, a secondary
metabolite. The present result could be substantiated
with the result of Abdul Jaffar Ali (2004) who reported
the presence of tyrosine derived secondary metabolite in
P. nigra. Low level of vanadium was also reported in
Ciona intestinalis with 100 ppm (Goldberg et al., 1951)
and in Molgula mortenseni with 54 ppm (Carlisle, 1954).
Millar (1954) also pointed out that the members of the
families Ascidiidae and Perophoridae from the order
Phlebrobranchia and several species in the order of
Aplausobranchia were found to accumulate
comparatively large vanadium concentration.
Annual mean concentration of cadmium in the
test and the mantle was 0.98 and 5.42 ppm respectively.
Maximum level was found to occur during July, 2011
and September, 2010 in the test (1.49 ppm) and the
mantle body (11.34 ppm) respectively whereas the
minimum concentration was during March, 2011 for the
test (0.063 ppm) and May 2011 for the mantle body
(0.35 ppm) (Fig 3). The concentrations of Cd observed in
P. nigra are higher than the reported values for other
ascidians such as Botryllus schlosseri 2.7 ppm
(Leatherland and Burton, 1974) and Ascidiaceae sp 0.2
ppm (Eustace, 1974). Bindu et al., (2007) noticed mean
cadmium level of about 4.1, 2.09, 1.15 and 0.09 ppm in
clams collected at Palk Bay, Bay of Bengal,
Rameshwaram and Sethukarai situated along the Gulf of
Mannar respectively whereas, higher concentration was
observed by Rejomon et al., (2008) in zooplankton with
28.1, 25.7 and 22.6 ppm in Chennai, Kanniyakumari and
Cochin respectively.
Similar to the cadmium, lead was also more in
the mantle body with mean value of 20.78 ppm and as in
the case of cadmium, the minimum concentration (12.75
ppm) was observed in the test. Maximum level in the
test (3.49 ppm) and the mantle body (5.68 ppm) was
during monsoon; whereas the minimum level was during
May, 2011 for the test (4.59 ppm) and the mantle body
(0.32 ppm) respectively (Fig4). The mean concentration
of Pb was significantly higher as compared to other
simple ascidian Styela plicata with 35 ppm (Matida and
Kumada, 1969) while, in other filter feeder,
Crassostrea madrasensis inhabiting Ennore, very low
accumulation was observed by Joseph and Srivastava
(1993) with 5.2 ppm.
Mercury was accumulated in both the test and
mantle body. Maximum concentration of mercury in the
test (20.7 ppb) and the mantle body (34.77 ppb) was
Ali et al., 2013
Journal of Research in Animal Sciences (2013) 2(1): 060-067 064
Figures 3. Seasonal variations in the accumulation of cadmium in the test and mantle body of
Phallusia nigra at Vizhinjam Bay.


during September, 2010 (Monsoon), while the minimum
was during premonsoon for the test (16.7 ppb) and the
mantle body (19.77 ppb) respectively (Fig 5). ANOVA
for the concentration of these metals between test and
mantle body showed significant difference (P<0.001).
Comparatively, mercury was found to be accumulated
lesser than the other ascidians such as Styela clavawith
0.13 ppm and B. schlosseri with 0.57 ppm (Leather land
and Burton, 1974). This clearly indicates that P. nigra
has significantly higher ability to accumulate trace
metals.
High concentration of these metals was found to
be during monsoon season at the study area and this
could be due to the fact that these metals might be from
land run-off and river run-off and these waters might
bring heavy load of metals through various sources. The
rain water usually has the tendency to chemically break
down the soils and rocks, and so it may contain higher
concentration of metals. Further leaching of metals from
soil, and wash off of metal based pesticides may also
contaminate the land and river run-off as stated by
Horvath et al., (1972). These run-off waters might
pollute the coastal water directly leading to the
accumulation of metals in P. nigra through the filter
feeding mechanism. Lower concentration observed
during premonsoon season might be due to decreased
inflow of metal enriched run-off, reduced free ions in
water effected by high salinity and also due to the
dilution of metals concentrations could be possible due
to surface currents and upwelling as stated earlier by
Nair et al., (1997).Application of one way ANOVA for
the concentration of these metals between test and
mantle body showed significant differences (p<0.005).

CONCLUSION
In conclusion, the results of the present study
clearly demonstrate that P. nigra examined showed a
great ability to accumulate concentrations of metals
several thousand times than those detected in sea water.
Metal concentrations recorded in this ascidian could
effectively be used as good reference material for
monitoring metal contamination in Indian sea waters.
Although the ascidian species studied in the present work
and suggested as biomonitors present numerous
advantages, more information and more studies on
ascidians are necessary to clarify accumulation patterns.
Nevertheless, this study can be looked upon as a starting
point for further investigations. The most effective
Ali et al., 2013
065 Journal of Research in Animal Sciences (2013) 2(1): 060-067
Figures 4. Seasonal variations in the accumulation of lead in the test and mantle body of
Phallusia nigra Vizhinjam Bay.
biomonitoring approach is to use several species of
ascidians representing different trophic levels in
combination with chemical and hydrologic
measurements.

ACKNOWLEDGMENT
The authors are thankful to OPMEC-CECRI,
Harbour area, Tuticorin for Atomic Absorption
Spectroscopic analysis of the heavy metals. The first
author acknowledges L.M. Muneer Ahmed Sahib,
Secretary and Dr. K. Prem Nazeer, Principal, Islamiah
College, Vaniyambadi for their great enthusiasm and
wise advice.

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