Advances in Biological Chemistry, 2013, 3, 549-552 ABC Published Online December 2013 (
A preliminary effort to assign sponge (Callispongia sp) as
trace metal biomonitor for Pb, Cd, Zn, and Cr, an
environmental perspective in Hative gulf waters Ambon
Netty Siahaya1*, Alfian Noor2, Nunuk Sukamto2, Nicole de Voogd3
1Department of Chemistry, Pattimura University Kampus Poka-Unpatti, Ambon, Indonesia
2Department of Chemistry, Hasanuddin University Kampus UNHAS Tamalanrea, Makassar, Indonesia
3Netherlands Center for Biodiversity Naturalis, Leiden, The Netherlands
Email: *Anethsia@gmail.Com
Received 4 November 2013; revised 5 December 2013; accepted 16 December 2013
Copyright © 2013 Netty Siahaya et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
The aim of this investigation is to estimate metal con-
centration distribution, Pb, Cd, Zn, and Cr, in Cal-
lispongia sp, sediment, and water column in Hative
waters of Ambon bay. After sampling, analytical pro-
cesses were conducted by using a standard method
and measured by ICP-OES (Inductively Coupled Plas-
ma Optical Emission Spectroscopy). The results show-
ed that Zn concentration was highest, 0.231 mg/kg
dry weight (ppm) while in sediment and water the
numbers were 1.180 and 0.790 ppm respectively. In
the meantime, functional group studies through Fou-
rier Transformed Infra Red spectroscopy revealed
that OH-group (3421 cm1), Conjugated Double Bond
(1635 cm1) and Ether Groups (1049 cm1) of callis-
pongia sp were found. Also, the appearance of wave-
number 2926 cm1 clearly indicates the existence of
–CH sp3 group. Combining both results may guide us
to establish a relation between metal concentration and
organic substance transformation made during inter-
action inside species. Thus, trace metal investigation
can be a choice in doing rapid assessment and bio-
monitor for the quality of coastal marine life. Fur-
thermore, by more extensive studies, this connection
could be developed and proposed as a low cost meth-
od for EIA of metal pollution in coastal zone, particu-
larly in coral reef system.
Keywords: Callyspongia sp; ICP-OES; FTIR; Metal
The capital of Maluku province is surrounded by Ambon
waters in the area about 187 thousand square kilometers,
17% slope, and relatively rugged land area. Its land use
will have an impact on the ecological pressures to Ambon
Bay waters [1]. Ambon Bay waters have multiple functions,
namely as a regional fisheries and aquaculture, police
seaport and Navy bases, PELNI ship port, and out of the
traditional boats Ambon and ferry crossing pier, fishing
port, Pertamina, ship repair dock space, recreation areas
and sports, and electric power by PLN, so that the waters
of Ambon Bay are very susceptible to environmental changes
because each activity will produce many metal wastes.
Island of Ambon is encircled by fringing type coral reefs
which grow lengthwise along the shoreline on the north
and south bay [2].
It is generally recognized that sponges are the animals
that always associated with coral reefs. Sponges, in
feeding, rely mostly on a constant flow of waters bringing
its dissolved organic matters as well as particulate non
organics such as metal associated compounds. The
sponges also have long been a center of attention from
scientists in different countries to look at the possibility
using them as metal bioindicator due to its capacity to
accumulate metals [3-6] without sacrificing its growth
and survival rate. Copper, lead, and vanadium have been
studied and showed a sponge capacity in absorbing them
in high concentration [4]. Also, Petrossian tertudinaria
species have been used as a biomarker for the detection of
heavy metals in inshore areas (0.5 - 1 km) and offshore (5
- 7 km) in the Gulf of Mannar, India. In fact, sponge heavy
metal concentration of inshore was about 64 times higher
than the offshore [3].
The ability to accumulate heavy metals in the sponge
is very important to be known as one of the guidelines in
determining the status of water pollution in an area that
has a coral reef ecosystem, in which the region is the
*Corresponding author.
N. Siahaya et al. / Advances in Biological Chemistry 3 (2013) 549-552
habitat of a living sponge. This is in line with the
opinions [3,4,9] that the metal content in aquatic biota
generally increases over time because the metal is ex-
pected, so the presence of a sponge in water Hative cycle
can be used to determine the heavy metals Pb, Cd , Cr,
and Zn in the sponge type callispongia sp, sediments and
water in the waters Hative Ambon bay.
Research materials are: sponges from Hative waters in
Ambon bay; Acetone (Merck), HNO3 (p.A), double dis-
tilled water and Whatman filter paper. Sponge samples
were collected by diving, cleaned and then placed in a
plastic bag and put in the ice box. 0.5 grams of sample
was put in beaker glass, add 5 mL HNO3 and then heated
at 150˚C for 2 hours. After being cooled at room tem-
perature, sample put in 25 mL volumetric flask, match
the volume with double distilled water and filtered with
Whatman paper and solution is ready to be analyzed by
ICP-OES Perkin Elmer 3000. One litre of water sample
was taken at the bottom, and immediately filtered with
filter paper of cellulose nitrate (0.45 μ) after previously
washed with 1N HNO3 and then preserved in HNO3 5%.
250 ml water sample is inserted in a Teflon separating
funnel, then extracted with APDC- NaDDC/MIBK. The
organic phase was extracted again with 5% HNO3 solu-
tion, filtered back, and ready for analysis by ICP-OES
Perkin Elmer 3000.
For sediments, they were taken from the bottom with a
Van Veen Grab sampler, stored in polyethylene bottles
and taken in laboratory put in a Teflon beaker and dried
in an oven at a temperature of 105˚C and after drying,
rinsed 3 times with double distilled water then dried
again. A total of 5 g sample was destructed in Teflon
beaker with solution of HNO3/HCl (1:3) at 100˚C for 8
hours. After that, the solution was filtered, and the fil-
trate is ready to be analyzed by ICP-OES Perkin 3000.
3.1. General Situation of Sampling Locations
Sampling was conducted on July 4, 2011, about noon
under cloudy weather in Hative waters (Figure 1) sur-
rounded by population settlement, estuaries, navy com-
plex, harbour, oil depot of Pertamina, and sago plantation.
Callispongia sp, ca 50 grams were taken under physico-
chemical as shown in Table 1. The data clearly shows
that its water quality fits with environmental conditions
in general where sponges grow in tropical and sub-
tropical conditions with vertical distribution on coral
reefs at low tide up into the area of approximately 50
meters [6]. Also, water content, ash, and biomass of Cal-
lispongia sp, represent an integral part of metal content
in a biological sample [9].
Accumulation of Pb, Cd, Cr and Zn Callispongia sp
can be seen in Table 2 as a function of organ as skeleton
and tissue. Also determined were water and sediment
around sponge. Logically before entering into cellular
level of sponge, metals will be firstly existed in water
and sediment It seems obvious that each element shows a
different partitional pattern where cadmium as the least
amount remain largely in sediment (86.4 %). Oppositely,
chromium was almost all adsorbed by cellular sponge. It
was also clear that majority of elements have stayed in
cellular level.
As for zinc as an essential element, its highest con-
centration revealed that the source might not only come
from industrial by products but also from excretion of
living system in marine environment. So the total zinc in
this Callispongia sp apparently not an abnormal case
especially if one looks at a fraction remaining in sedi-
ment (54.1%) and consumption level of skeleton and
tissue on zinc. Generally the highest metal concentration
in sediments is influenced by several processes like
sedimentation, flocculation, precipitation, and adsorption
[5,6]. Another important parameter is bioconcentration
factor (BCF) in measuring the capability of organisms to
accumulate metals from environment into its tissue. Ac-
cording to Abdullah et al. 2007, BCF can be estimated
by comparing metal absorption between in sponge tissue
and in water (BCF sw) or sediment (BCF ss).
The ability of organisms to accumulate metals from
the environment into the tissues of the body can be cal-
culated using the bioconcentration factor (BCF). BCF
value can be obtained by comparing the ability of organ-
isms (e.g. sponges) to absorbsi metals from water and
sediment. Therefore there are two BCF values, BCF
sponge-sediment (BCFs-s) and BCF-water sponge (BCFs-
w). BCFs-s is the value of the ratio between the concen-
tration of the metal absorbed into the sponge tissue with
metal concentrations in the sediment, while the BCF sw
is the value of the ratio between the concentration of
metal that accumulates into the sponge tissue metal con-
centrations in water [8] as found in table-3 below.
From the result in Table 3 one can find the highest
value for BCF is Cr either in water or in sediment. and
this indicated that for the case of the location and sponge,
Cr is the most appropriate element to be assigned as a
bioindicator or biomonitor for Callispongia sp compared
to other metals. Some results from several investigations
have used the same method [4,5,7].
To assess the possible chemical bonding occurs be-
tween organic molecules in sponge and metal, an FTIR
analysis was carried out to look at the key functional
groups that may link to bond formation with metals.
FTIR spectrum of Callispongia sp sample can be seen in
Figure 2.
According to Terada et al. (1983) the interactions that
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N. Siahaya et al. / Advances in Biological Chemistry 3 (2013) 549-552
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Figure 1. Sampling location is in the waters of Hative Besar municipal.
Figure 2. Fourier Transformed Infra Red (FTIR) spectrum of Callispongia sp tissue sample.
Table 1. Physicochemical conditions in Hative and sponge
Callispongia sp.
Table 3. BCF s-s and BCF sw of Pb, Cd, Zn, & Cr calculated
from Callispongia sp.
Site Waters conditions Content (%)
(˚C) pH Salinity
(0/00) Water Ash Biomass
28 6.8 30 78.74 87.00 16.90
Metal BCF ss BCF sw
Pb 0.32 0.41
Cd 0.52 10.0
Zn 0.09 0.13
Cr 262.0 262.0
Table 2. Metal content (ppm) in the sponge Callispongia sp.
Metal Total
Skeleton + Tissue Skeleton Tissue WatersSediment
Pb 0.039 0.008 0.031 0.0750.098
Cd 0.012 0.002 0.01 0.0010.019
Zn 0.213 0.109 0.104 0.7901.180
Cr 0.280 0.018 0.262 0.0010.001
have occured between active functional groups of orga-
nic molecules can be described as the behaviour of Lewis
acid-base interaction to form a complex chemical struc-
ture. In case of metal adsorption in a solution system,
these reactions may be generalized as follow :
 
N. Siahaya et al. / Advances in Biological Chemistry 3 (2013) 549-552
 
3.2. Where GH Is a Functional Group and M Is
a Divalent Metal Ion Z
The FTIR results showed that functional groups found in
Callispongia sp are OH (3421 cm1), the group of
conjugated double bonds (1635 cm1), and ether groups
(1049 cm1). While the wave number 2926 cm1 region
of the spectrum shows the possibilities that come from
sp3 CH orbital. These are functional groups that have
possibility to bind metals thus potential active groups to
catch metals.
It can be concluded that Callispongia sp may be assigned
as a pollutant indicator for metal especially element chro-
mium which is very toxic in certain forms and valences.
A further investigation will be needed to explore sponge
potential as the metal pollution bioindicator in approp-
riate locations particularly in eastern island littoral re-
gions where coral reef is found predominantly.
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