A Pilot Study on Effect of Copper and Cadmium Toxicity in

Tilapia Mossambicus
Keywords:
Toxicity, Aquaculture, Trace metals, Tilapia mossambicus.
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Dates:
Received: 10 Mar 2012 Accepted: 19 Apr 2012 Published: 13 Jun 2012
Article Citation:
Anushia C, Sampath kumar P and Selva Prabhu A.
A Pilot Study on Effect of Copper and Cadmium Toxicity in Tilapia Mossambicus.
Journal of Research in Animal Sciences (2012) 1: 020-027
An International Online Open Access
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Journal of Research in Animal Sciences
ABSTRACT:

Cu and Cd is trace element for most organisms including fish, but above
certain limit Cu and Cd will be toxic. The present study was conducted to evaluate the
toxic effect of Cu and Cd on Tilapia mossambicus via estimating the acute 96h median
lethal concentration (LC
50
) value. A total 120 number of Tilapia mossambicus
fingerlings were subjected to 12 numbers 20-L aquaria. Fish were exposed to 0.0, 2.0,
4.0, 6.0, 8.0 and 10.0mg Cu and Cd/L for 4 days. Each dose was represented by two
aquaria. Fish was daily observed and dead fish were removed immediately. The data
obtained were evaluated using Behrens-Karber’s Method. The 96 h LC
50
value of Cu
for Tilapia mossambicus was calculated to be 6.0mg Cu/L with Behrens-Karber’s
Method. The 96 h LC
50
value of Cd for Tilapia mossambicus was calculated to be
4.8mg Cd/L with Behrens-Karber’s Method. The behavioral changes of
Tilapia mossambicus were primarily observed. It could be concluded that
Tilapia mossambicus species slightly sensitive to Cu and Cd when compare both metal
cadmium is more toxic than copper for the fish species.
Authors:
Anushia C, Sampath
kumar P and Selva
Prabhu A.


Institution:
Centre of Advanced Study in
Marine Biology, Faculty of
Marine Sciences, Annamalai
University, Parangipettai-
608 502, Tamil Nadu, India.



Corresponding author:
Anushia C.



Email:
anushiaanubiotech@gmail.com.












Web Address:
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INTRODUCTION
Fishes are widely used to evaluate the health of
aquatic ecosystems because pollutants build up in the
food chain and are responsible for adverse effects and
death in the aquatic systems (Farkas et al., 2002; Yousuf
and El-Shahawi, 1999). The studies carried out on
various fishes have shown that heavy metals may alter
the physiological activities and biochemical parameters
both in tissues and in blood (Basa and Rani, 2003; Canli,
1995; Tort and Torres, 1988). Tilapia is distinguished by
its adaptation to living in fresh, brackish and nearly
saline water, and can survive in partially polluted water
(Zyadah, 1995). It is less sensitive to most toxic
substances than most other aquatic species. Any toxicant
that affects Tilapia would most likely be toxic to other
aquatic organisms (Murungi and Robinson, 1987). The
toxic effects of heavy metals have been reviewed,
including bioaccumulation (Waqar, 2006; Adami et al.,
2002; Rasmussen and Anderson, 2000; Rani, 2000;
Aucoin et al., 1999). Marine coastal ecosystems could
therefore be endangered by pollutants, such as heavy
metals, pesticides and antifoulants that could be easily
detected in the water column or in the sediment of
harbors and estuaries (Castillo et al., 2006;
Antizar, 2008; Bellas, 2005).
Heavy metals are considered the most important
form of pollution of the aquatic environment because of
their toxicity and accumulation by organisms, such as
fish (Emami Khansari, Ghazi-Khansari, and Abdollahi,
2005). Besides, the dangers involved from the presence
of metals in the environment derive not only from their
persistence and toxicity, but also from the remarkable
degree of bioaccumulation they undergo through the
tropic chain, thus becoming serious danger to man
(Bishop, 2000). Heavy metals such as cadmium, copper
and lead are found in most of the industrial, mineral
exploration and other miscellaneous anthropogenic
effluents. In Nigeria, these effluents are indiscriminately
discharged into natural waters, thereby contaminating
aquatic ecosystem (Fafioye et al., 2002).
Trace metals, such as Cu, Ni, Fe, and Cd are
accumulated over time in higher concentrations in fish
liver, gills and muscles (Taylor et al., 1985). Besides the
direct impact of heavy metals in fish, the synergistic
action of some hydrological variables and nutrients to
fish was found to enhance heavy metal toxicity in fish
(Bu-Olayan and Thomas, 2005; Franco et al., 2006). The
acute trace metal toxicity levels in fish exposed from
24 h to 96 h was statistically tested using Profit program
(USEPA, 1993) by various investigators (Abel and
Axiak, 1991; APHA, 1992; Wayne and Ming, 1998;
Franco et al., 2006). Heath (1987) described varied
pattern of inorganic pollutant bioaccumulation in
different fish tissues such as liver, muscles and gills.
Toxicity tests using aquatic organisms play an
important role in the development of proposals for
environmental management and protection, especially
for the aquaculture environment (Wall and Hanmer
1987; Hoi, 2004. In addition, it is an important step to
detect the levels of toxicants to be used in the
experimental studies of the accumulation and effect of
these toxicants to the marine organisms. There are many
studies concern with the toxicity of cadmium on
vertebrates and invertebrates (Rasmussen and Andersen,
2000, Adami et al., 2002 and Filipovic and Raspor,
2003). These metals are readily seeped by industries into
our waters daily, thereby increasing their accumulation
level. Therefore, it is necessary to study the toxicity of
cadmium and copper with a view to predict their level of
toxicity to Tilapia mossambicus.

MATERIALS AND METHODS
Fish management
Apparently healthy Tilapia mossambicus
(3.5±0.2g) were obtained from local fish farm Pinnalore,
Cuddalore Dist, Tamilnadu, India. Prior to the
experiment, fish were acclimatized for 2 weeks in 12
numbers 40-L glass aquaria under laboratory conditions
Anushia et al., 2012
021 Journal of Research in Animal Sciences (2012) 1: 020-027
(natural photoperiod 11.58-12.38 h); 10 fish per each
aquarium. The continuous aeration was maintained in
each aquarium using an electric air pumping
compressors.
Analysis of the water physico-chemical variables
Temperature
The atmospheric temperature and surface water
temperature were noticed with the help of a degree
Celsius thermometer.
Salinity
Salinity was recorded using a hand
Refractometer (Atago, Japan).
Hydrogen-ion concentration (pH)
Water pH (Negative logarithm of hydrogen ion
concentration) was noted by a calibrated pH pen
(pH Scan 1 Tester-Eutech Instruments, Singapore).
Dissolved oxygen
Dissolved oxygen was measured by using a
modified Winkler’s titration methods described by
Strickland and Parsons (1972).
Experimental procedures
The heavy metal Cu in the form of Copper
chloride anhydrous (Merck, Mumbai, India) and Cd in
the form of Cadmium chloride (Merck, Mumbai, India)
was used in the present study. The acute toxicity test was
performed for 4 days in which two replicates of seven
different Cu and Cd concentrations (0, 2, 4, 6, 8 and
10mg/L) were used (10 fish for each aquarium). At 24,
48, 72, and 96 h, fish dead were counted in the different
Cu and Cd concentrations along with the control group.
In this study, the acute toxic effects of Cu and Cd on
Tilapia mossambicus were deter mined by
Behrens-Karber’s method using the following formula
(Klassen, 1991):
LC
50
= LC
100
∑A x B / N as mg/L;
Where LC
50
and LC
100
indicate the lethal doses for 50%
and 100% of the tested fish. Value ‘‘A” gives the
differences between the two consecutive doses, ‘‘B” the
arithmetic mean of the mortality caused by two
consecutive doses and ‘‘N” the number of tested fish in
each group. The dead fish were removed immediately.

RESULTS
The data obtained from the acute toxicity test of
copper for Tilapia mossambicus revealed that the Cu
toxicity increased with increasing concentration or
exposure time. The numbers of dead fishes in relation to
different Cu concentrations (2, 4, 6, 8 and 10mg/L) were
assessed and counted during the exposure in different
time intervals at 24, 48, 72 and 96 hours. Then the dead
fishes were removed immediately from the culture tanks.
No mortality was observed during the 96 h at control
(0.0mg Cu/L) and 100% mortality rate was achieved
only at 10mg Cu/L
During the toxicity tests, the temperature, salinity
and ph of the test water remained fairly constant at
28.5±1.5°C, 3.1±2.6mg/l and 7.5±0.4 respectively, while
dissolved oxygen was higher than 5.84±0.72mg/l. There
Anushia et al., 2012
Journal of Research in Animal Sciences (2012) 1: 020-027 022
Cu dose(mg/L) No. of exposed fish
No of dead fish
Overall deaths within 96 h A B AB
D1 D2 D3 D4
0 10 0 0 0 0 0 0 0 0
2 10 0 0 0 1 1 2 0.5 1
4 10 0 1 1 1 1 2 1.0 2
6 10 1 4 5 5 5 2 3.0 6
8 10 6 8 8 8 8 2 6.5 13
10 10 8 10 10 10 10 2 9.0 18
∑AB =40
Table 1. The cumulative mortalities and acute 96 h LC
50
of Cu in Tilapia mossambicus
according to Behrens-Karber’s method (Klassen, 1991).
Where A = differences between the two consecutive doses and B = arithmetic mean of the mortality caused
by two consecutive doses. 96 h LC
50
= LC
100
- ∑ (A x B)/N = 10 – 40/10 = 6. Ppm.


was 100% survival at initial exposure in the different
concentrations, but the survival rate started declining
with an increase in concentrations and time of exposure.
Effect of Copper
When exposed to copper, Tilapia mossambicus
recorded 80% and 50% mortality in 8mg/l and 6mg/l of
Cu respectively at 96h duration. The lowest
concentration (2.0mg/l) produced 10% mortality at 96 h
in Tilapia mossambicus. When compare to control
mortality was inhibited at 2mg/L in 48 hrs. The 96h
LC
50
value (6.mg/L) of T. mossambicus was determined
based on measured concentration of copper with the
Behrens-Karber’s method (Table 1).
Effect of Cadmium
In Cadmium exposure percentage of mortality at
96 h was 90% in 8mg/l and 60% in 6mg/l of Cd, while
30% mortality occurred in 2mg/l and 4mg/l at 96 h.
Tilapia mossambicus had 100% mortality in 10mg/l. The
96h LC
50
value was estimated to be 4.8mg/L with the
Behrens-Karber’s method (Table 2).

DISCUSSION
Toxic effect on the fish in the present study and
toxicity increased with increased concentration. The
observed increasing state of inactivity with both
increasing concentrations and exposure period agree with
the report of Ayoola, (2008a). The present investigation
showed big differences of both toxicity and
bioaccumulation rate among the aquatic organisms. A
clear variation in LC
50
and acute toxicity in tested
organisms were evident. 96hr LC
50
of Cu was 6.0mg
Cu/l, while in Cd was 4.8mg Cd/l. Other research
reported lower Cu concentrations 96hr- LC
50
for marine
crustaceans as; 0.017mg Cu/l for Acartice tansa;
0.049mg Cu/l for Cancer magister and 0.1mg Cu/l for
Homarus americanus (Martin et al., 1981 and Mance,
1987).
The effect produced by both metals coupled is
less than the effects produced by individual metal. This
may attributed to substitution and competition between
Cu and Zn for available sites during protein synthesis as
suggested by Bryan, (1971) and Abdel-Moati and Farag
(1991). The variety degree is related to kind of species,
its sensitivity and physiological responses to pollutants.
And their uptake and depuration rate of heavy metals
(Salanki and V. -Balogh 1985; Salanki and V. -Balogh
1989). El- Gindy, et al., (1991) recorded 24h LC
50
for
mol lusks Bi omphal ari a al exandri na and
Bulinus truncatus of Cu and Zn toxicity as 1.38, 0.99 and
54, 40 ppm, respectively. The 96hr LC
50
value for Cu in
L, balteni was 0.9 ppm (Abdel-Moati & Farag 1991), but
in Mugil fry was 1.3 ppm (El-Rayis and Ezzat 1984).
The 96h LC
50
of Zn in L. bolteni was 58 ppm
(Abdel-Moati and Farag 1991), while that for
Portunus pelagicus was 100 ppm, (El-Rayis and Ezzat
(1984). However T. zillii have the ability to live in
Cd dose (mg/L) No. of exposed fish
No of dead fish
Overall deaths within 96 h A B AB
D1 D2 D3 D4
0 10 0 0 0 0 0 0 0 0
2 10 0 1 2 3 3 2 1.5 3
4 10 1 2 3 3 3 2 3.0 6
6 10 2 3 4 6 6 2 4.5 9
8 10 4 7 8 9 9 2 7.5 15
10 10 6 10 10 10 10 2 9.5 19
∑AB =52
Table 2. The cumulative mortalities and acute 96 h LC
50
of Cd in according to
Tilapia mossambicus Behrens-Karber's method (Klassen, 1991).
Where A = differences between the two consecutive doses and B = arithmetic mean of the mortality caused
by two consecutive doses. 96 h LC
50
= LC
100
- ∑ (A x B)/N = 10 – 52/10 = 4.8ppm.
This value was estimated to be 4.8mg/L with the Behrens–Karber’s method (Table 2).
Anushia et al., 2012
023 Journal of Research in Animal Sciences (2012) 1: 020-027
polluted areas for long time than other species of fish
(Zyadah, 1999). The actual and back calculated LC
50
of
Cu, Zn. and Cd values for the experimental species
during the exposure periods showed a close concordance.
Other results in the world showed different LC
50
of Cu,
Zn, and Cd values, where flounder fish exposed to 0.1 to
10 mg Cd/l for 15d (Larsson et al., 1976); Juvenile
striped bass was exposed to 0.01 mg Cd/l for 120d
(Dawson et al., 1977) and juvenile of shrimps
Penaeus duorarum exposed to 5 mg Cd/l for 96hr
(Nimmo et al., 1977). The rate of bioaccumulation of
heavy metals by fish and shrimp appeared within a wide
range. The bioaccumulation factor of Cd by Mysis sp.
was 1215 times more than control concentration after
48hr exposure, and reaches 858 times in T. zillii after
356hr exposure. Other studies in USA showed the
average residues of Cd in some invertebrate species to
reach approximately 1000 to 9000 times greater than
correspond control concentration after 28d exposure
(Spehar et al., 1978).
In the present study, it was observed that exposed
Tilapia mossambicus to various concentrations of
cadmium and copper were weakened progressively with
time prior to mortality. Similarly, the toxic effect of the
metals produced molting in the fish at a faster rate than
control. These facts, therefore, affirm that heavy metals
can cause physiological stress and dysfunction in
crustaceans (Gao and Zou, 1995).
The observed increasing state of inactivity with
both increasing concentrations and exposure period agree
with the report of Ayoola, (2008a). The results of
toxicity test indicated that the ionic form of Cu is more
toxic than the ionic form of Cd to Mugil seheli, and the
fingerlings are more sensitive to copper toxicity than that
of cadmium. Denton and Burdon-Jones (1986);
Cui-Keduo et al., (1987). Spehar et al., (1978) reported
that the 96 h LC
50
of Cd for flag fish,
Jordanella floridae, was 2.5mg /l. Hamed, (2002) found
that the 72 h LC
50
of Cd for Mugil seheli was 4.87mg/1.
El-Moselhy, (2001) stated that toxicity of Cd to
Mugil seheli decreased with increasing the exposure time
and the recording LC
50
values were 12.34, 8.92, 6.01 and
3.45mg/l for 24, 48, 72 and 96 hours, respectively. The
96 h LC
50
values of copper was 1.83 ppm for fish
Etroplus maculaus reported by Gaikwad, (1989).
Taylor et al., (1985) reported LC
50
values of about 0.3 to
50mg Cd/1. While 96 h LC
50
of Cu ranged from 0.2 to
3mg/1 for various marine fish and crustaceans (Bryan,
1971). Pagenkopf, (1986) studied the toxicity of copper,
cadmium, lead and zinc to fishes.
The values worked in the present experiment as
safe concentrations of Cu and Cd to reach LC
50

concentration and total mortality dose to aquatic
organisms, these are of great practical utility for
regulating and controlling the pollution limits in the
water resources by those pollutants and to regulate their
discharge to near-by water for protect the life within the
aquatic environment.
The susceptibility of fish to a particular heavy
metal is a very important factor for LC
50
values. The fish
that is highly susceptible to the toxicity of one metal may
be less or non-susceptible to the toxicity of another metal
at the same concentration of that metal in the milieu.
Similarly, the metal which is highly toxic to one
organism at low concentration may be less or non-toxic
to other organism at the same or even higher
concentrations with two juvenil Brazilian indigenous
fishes which showed that both species were more
sensitive to copper and cadmium found that with
Daphnia pulex the order of toxicity of different metals
was Cu>Cd>Ni.

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Anushia et al., 2012
027 Journal of Research in Animal Sciences (2012) 1: 020-027