Acute toxicity bioassay of three different heavy metals copper (Cu), lead (Pb) and cadmium (Cd) for the freshwater catfish, Clarias batrachus were conducted for evaluation of the toxicity. The 96h&nbsp;LC50 values recorded for Cu, Pb and Cd were 15 ppm, 29 ppm and 35 ppm respectively. The results of this study indicate that the order of toxicity of these heavy metals is Cu&gt;Pb&gt;Cd in short duration acute toxicity experiments. The toxicity was found to be dependent on dose and duration of experiment for all these heavy metals. Cu was found to be much more toxic than Pb and Cd to C. batrachus

Gandhewar, S.S.

Sitre, S.R.

Zade, S.B.

Journal of Applied and Natural Science

   JANS Journal of Applied and Natural Science 6 (1): 117-120 (2014) Assessment of toxic potential of three different heavy metals to Clarias  batrachus (Linn.) utilizing static acute bioassay  S.S. Gandhewar1*, S.B. Zade2 and S.R. Sitre3 1J.B. College of Science, Wardha (M.S.), INDIA  2P.G. Department of Zoology, R.T.M. Nagpur University Campus, Amravati Road, Nagpur- 440033 (MS), INDIA 3N.S. Science and Arts College, Bhadrawati, District Chandrapur-442902 (M.S.), INDIA *Corresponding author. E-mail: ssgandhewar2007@rediffmail.com   Received: November 12, 2013; Revised received: February 3, 2014; Accepted: April 4, 2014 Abstract:  Acute toxicity bioassay of three different heavy metals copper (Cu), lead (Pb) and cadmium (Cd) for the freshwater catfish, Clarias batrachus were conducted for evaluation of the toxicity.  The 96h LC50 values recorded for Cu, Pb and Cd were 15 ppm, 29 ppm and 35 ppm respectively. The results of this study indicate that the order of toxicity of these heavy metals is Cu>Pb>Cd in short duration acute toxicity experiments. The toxicity was found to be dependent on dose and duration of experiment for all these heavy metals. Cu was found to be much more toxic than Pb and Cd to C. batrachus. Keywords:   Acute toxicity,  Cadmium, Copper, LC50,  Lead ISSN : 0974-9411 (Print), 2231-5209 (Online)  All Rights Reserved © Applied and Natural Science Foundation  www.ansfoundation.org INTRODUCTION The human efforts to increase production of food,  energy and luxury things to enhance the standard of  living resulted in increased industrialization,  urbanization and rapid development. Heavy metals and other pollutants do not enter the aquatic environment as single entity, but are released in complex form (Olojo et al., 2005 ). The concern for environmental protection with respect to the aquatic environment arises from the fact that, various chemicals are  discharged into surface water bodies which  subsequently disturb their fragile ecology. As a result, several sensitive species are lost from the nature  resulting in depletion of valuable biodiversity. Various non-biodegradable xenobiotics present in the industrial effluents get bound to sediments, animal and plant tissues and continue to exert their toxic effect due to slow release of toxicants (Adham et al., 2002; Olojo et al., 2005). Aquatic ecosystems are sensitive to exposure of toxic contaminants and heavy metals. Among the aquatic  organisms, fish is most vulnerable to these contami-nants (Alinnor, 2005). Lead is a naturally occurring heavy metal present in the earth’s crust, rocks, soil and water. Most water borne lead is derived from human activities such as mining, smelting, coal burning,  cement manufacturing, use of gasoline, batteries and paint (Ramesh et al., 2009).  Cadmium is one of the most toxic and widespread heavy metal and is a  recognized carcinogen in mammals (Pruski and Dixon, 2002).  Cadmium has become the focus of global  research due to its toxicity to terrestrial and aquatic organisms while copper also acts as a potent toxin in aquatic organisms, when it enters into aquatic  ecosystems. In this context toxicity of Cu, Pb and Cd to freshwater catfish C. batrachus  was assessed in short duration acute toxicity experiments in order to  conserve and protect the freshwater catfishes in natural aquatic ecosystems. The significance of the present study lies in the fact of conservation and protection of  aquatic biota which is useful as a food factor for the man in long run which is to be protected on top  priority. MATERIALS AND METHODS Study material: 100 healthy fish  weighing about  40 - 45 gm. were procured from natural unpolluted sources reservoir and brought to the laboratory, exam-ined for any pathological symptoms, treated with 0.1% KMnO4 solution to avoid any dermal infection and acclimatized in dechlorinated tap water for 15 days.  The fishes were fed on minced goat liver and tubifex worms once in a day in morning hours. The test fishes were selected and maintained in separate aquaria  before actual experimentation.  Preparation of metal solution: The three different heavy metal salts viz. copper sulphate (CuSO4.5H2O), lead acetate (Pb(C2H3O2)2) and cadmium chloride (CdCl2) (A.R.Grade) were selected for toxicological experiments. Dechlorinated tap water which was stored in a large overhead tank for about ten days was used for conducting the toxicity experiments. The physico-chemical characteristics of the dechlorinated tap water were determined as per  standard methods 2008APPLIED    ANDNATURAL SCIENCEFOUNDATIONANSF118   (APHA, 2012). The details of the physico-chemical characteristics of water are given below:  Stock solution of the three heavy metals (copper  sulphate, lead acetate and cadmium chloride) and their dilutions were made according to standard guidelines  (OECD, 1981; APHA, 2012). Experimental design and toxicity bioassay: In order to determine the LC50 of CuSO4.5H2O, Pb(C2H3O2)2 and CdCl2 (A.R.Grade)  and their combination, method of Litchfield and Wilcoxon (1949) was followed.  Successive batches of 10 fishes were exposed to  different concentrations of heavy metals under  standard laboratory conditions after 24,48,72 and 96 hrs. duration. After specified time interval, the number of fish dead and alive were noted (Sprague, 1969). The experiments were carried out in glass aquaria of 100 L capacity in static laboratory conditions. RESULTS AND DISCUSSION   The percent mortality observed for each dose was  calculated and converted to probits by means of a  probit table. Dosage versus percent mortality was  plotted on the graph. The LC50 values and the 95% confidence limits of heavy metals, copper sulphate, lead acetate and cadmium chloride for fish, C. batra-chus  during 24, 48, 72 and 96 hrs are presented in table 1 . The toxicity tolerance of freshwater fish C. batrachus to CuSO4.5H2O, Pb(C2H3O2)2 and CdCl2 in the present investigation are dose and duration depend-ent. It was noticed that mortality increased with an increase in concentration. LC50 values of all the three heavy metals as well as their combination showed the susceptibility of catfish to lethal concentrations in acute short term exposure.  Higher percent mortality occurred in the fish Clarias Table 1. LC50, 95% confidence limits and slope function values of  Cu to C. batrachus in acute toxicity bioassay. Exposure  Period (hrs.) Parameter Values Copper Lead Cadmium 24 LC50 mg/L 25 42 54 95% confidence interval 20.40 – 30.60 33.60 – 52.50 44.32 – 64.00 Slope function y=15.35x – 14.28 y=13.16x – 12.5 y=14.04x – 20.71 Regression (R2) 0.979 0.963 0.969 48 LC50 mg/L 20 37 44 95% confidence interval 16.39 – 24.40 29.13 – 46.99 34.37 – 56.32 Slope function y=15.35x – 14.28 y=10.5x – 13.61 y=13.33x – 20 Regression (R2) 0.979 0.989 0.982 72 LC50 mg/L 17.50 34 40 95% confidence interval 14.30 – 21.35 26.56 – 43.52 34.18 – 46.80 Slope function y=13.21x + 2.857 y=11.33x – 22.22 y=11.78x – 21.78 Regression (R2) 0.910 0.960 0.958 96 LC50 mg/L 15 29 35 95% confidence interval 10.59 – 20.50 21.46 – 39.15 23.30 – 52.50 Slope function y=15.35x + 0.00 y = 9.66x – 21.66 y=11.78x – 21.78 Regression (R2) 0.932 0.934 0.958 S.N. Physico-chemical characteris-tics of dilution water Range 1 pH 7.2-7.4 2 Temperature  (oC) 24-28 3 Dissolved Oxygen (DO)  (mg/l) 6.9-7.1 4 Alkalinity (as CaCO3) (mg/l) 160-170 5 Total hardness (as CaCO3) (mg/l) 160-180 S.S. Gandhewar et al. / J. Appl. & Nat. Sci. 6 (1): 117-120 (2014) 119   batrachus, under investigation with increase in  concentration and exposure period, which confirms the similar observations recorded by researchers world-wide such as in case of Oncorhynchus mykiss, Salvelinus confluentus and Oncorhynchus tshawytscha (Finalayson and Verrue, 1982; Hansen et al., 2002), guppy, Poecilia reticulata (Yilmaz et al., 2004),  Cyprinus carpio (Muley et al., 2000; Dardenne et al., 2007), Nile tilapia, Oreochromis niloticus (Garcia et al., 2006) and Rohu, Labeo rohita (Dutta and Kaviraj, 2001).  In the present research work, Finney's Probit Analysis for evaluating the acute toxicity response gave 96 hour LC50 value for C. batrachus exposed to CuSO4.5H2O as 15 ppm, Pb(C2H3O2)2  29 ppm, CdCl2  35 ppm. From the derived LC50 values, it is quite clear that, the toxic effect increased with dose and duration. Our  results are in agreement with Yilmaz et al. (2004) who reported 96 h LC50 value of cadmium to guppy (P. reticulata). The other supporting studies are of Oryan and Nejatkhah (1997), Woodal et al. (1988) and Muley et al. (2000). Today, worldwide heavy metals are introduced into aquatic environment through industrial processes, soil leaching, mining activities, sewage disposal and  rainfall. The heavy metals are relatively toxic, even at fairly low concentrations and affect the survival of all other aquatic organ-isms. Stebbing and Fandino (1983) reported that, the adverse biological effects of heavy metals in the aquatic environment are mainly due to their complex nature. When the toxicant concentration in the water body is very high, it results in the death of fish. So, the death of an organism was taken as the end point of toxicological studies previously (Jones and Reynolds, 1997). However, Adams (1990) stated that sublethal concentrations of toxicant also induce sub-stantial changes in the biological organization of fish. The LC50 values derived from toxicological investiga-tions are highly useful in determining the sublethal concentration of a metal. Today, most of the  information on the effects of heavy metals on aquatic organisms is focused on short duration experiments  carried out at lethal concentrations. The information thus derived is not sufficient to assess the extent of damage. Hence, there is a need to carry out studies related to sublethal toxicity. Such studies are highly valuable in evaluating the sequence of events that  involve the response of the test animal to sublethal concentrations (Nobbs and Pearu, 1976; Perkin, 1979). So, in order to derive sublethal concentrations and to evaluate the response of the fish at sublethal concentra-tions, LC50 values are of prime importance in today's changing scenario of environmental pollution. Conclusion The present toxicological investigation clearly showed that the toxicity is dose and duration dependent for all the three heavy metals. The heavy metal Cu was much more toxic than Pb and Cd. The mortality increased with an increase in concentration and  LC50 values of all these heavy metals showed the susceptibility of catfish to lethal concentrations in acute short term  exposure. Such type of toxicological studies will be useful for conservation and protection of  aquatic  organisms and ultimately safeguarding the interest of man in long run as a  food supply.  REFERENCES Adams, S.M. (1990). Status and use of biological indicators for evaluating the effects of stress of fish. 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Assessment of toxic potential of three different heavy metals to Clarias batrachus (Linn.) utilizing static acute bioassay

Acute toxicity bioassay of three different heavy metals copper (Cu), lead (Pb) and cadmium (Cd) for the freshwater catfish, Clarias batrachus were conducted for evaluation of the toxicity. The 96h LC50 values recorded for Cu, Pb and Cd were 15 ppm, 29 ppm and 35 ppm respectively. The results of this study indicate that the order of toxicity of these heavy metals is Cu&gt;Pb&gt;Cd in short duration acute toxicity experiments. The toxicity was found to be dependent on dose and duration of experiment for all these heavy metals. Cu was found to be much more toxic than Pb and Cd to C. batrachus.</jats:p

Assessment of toxic potential of three different heavy metals to Clarias batrachus (Linn.) utilizing static acute bioassay

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