Int. J. Aquat. Biol. (2016) 4(1): 25-30 E-ISSN: 2322-5270; P-ISSN: 2383-0956 Journal homepage: www.ij-aquaticbiology.com © 2016 Iranian Society of Ichthyology Original Article Acute toxicity bioassay of the mercury chloride and copper Sulphate in Rutilus caspicus and Rutilus kutum Alireza Pourkhabbaz1, Borhan Mansouri2, 3, Mohammad Hosein Sinkakarimi4, Ghasem Rajaei*1, Robabeh Vajdi5 1Department of Environmental Sciences, Faculty of Natural Resource and Environment, Birjand University, Birjand, Iran. 2Student Research Committee, Kurdistan University of Medical Sciences, Sanandaj, Iran 3Environmental Health Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran 4Department of Environmental Science, Faculty of Natural Resource and Environment, Malayer University, Malayer, Iran. 5Department of Environmental Science, Faculty of Environmental, University of Tehran, Tehran, Iran. Article history: Received 12 July 2015 Accepted 22 January 2016 Available online 2 5 February 2016 Keywords: Toxicity LC50 Static bioassay Fish Abstract: The purpose of the present study was to determine the acute toxicity (LC50) of HgCl2 and CuSO4 in Caspian roach (Rutilus caspicus) and CuSO4 in Caspian kutum (Rutilus kutum). The Caspian roach LC50 values for HgCl2 at 24, 48, 72, and 96-hrs of exposure, were 0.64, 0.61, 0.42, and 0.28 mg L-1, respectively, and for CuSO4 were 11.55, 5.08, 2.49, and 1.47 mg L-1, respectively. The Caspian roach LC50 values for CuSO4 at 24, 48, 72, and 96-hrs of exposure, were 5.31, 4.17, 3.20, and 2.25 mg L-1, respectively. The results of this study showed that the toxicity of HgCl2 is higher than that of CuSO4 for the studied species. The mortality decreased with time, and most of the deaths were occurred during the first 24 hrs. Introduction Pollution pressures are associated with urbanization endangering the coastal ecosystems. The biota may be stressed by discharged specific point sources (e.g. sewage effluents and industrial wastes) and general non-point pollution (e.g. harbor activities, storm drainage and agricultural drainage). The aquatic ecosystems that receive pollutants are often important fishery and recreational areas (Pirbeigi et al., 2016; Nasrolah Pourmoghadam et al., 2015). It is therefore essential that techniques are established to monitor the pollutants that pose a danger to the aquatic biota and humans (Gopalakrishnan et al., 2008). Metals are an important group of the water pollutants that disturb the integrity of biochemical and physiological mechanisms in aquatic organisms, such as fishes. Among metals, copper and mercury are of special concern since they are considerably toxic to aquatic animals at ecologically relevant concentrations (Zhang et al., 2005; Allen et al., * Corresponding author: Ghasem Rajaei DOI: http://dx.doi.org/10.7508/ijab.2016.01.004 E-mail address: ghasem.rajaei19@yahoo.com 1994). Copper is a trace element that plays a fundamental role in the biochemistry of organisms, including aquatic organisms that can take it up directly from water (Grosell et al., 2003; Erickson et al., 1996). However, it can become toxic at high concentrations (Alquezar et al., 2008). Among metals, mercury is of special concern since it is considerably toxic to aquatic animals at ecologically relevant concentrations (Vieira et al., 2009; Oliveira et al., 2002). Mercury is considered one of the most dangerous metals in the aquatic environment (Goyer et al., 1995; Ribeiro et al., 1996), mainly its organic forms that can be biomagnified in trophic chains representing an increased risk for top predators (MacDougal et al., 1996), including humans consuming contaminated fishes. Chronic exposure and accumulation of these chemicals by aquatic biota can result in tissues that produce adverse effects not only in the exposed organisms, but also in human beings (IARC., 1990; Karthikeyan et al., 2007). Therefore, it is essential to study the detrimental 26 Pourkhabbaz et al./ Acute toxicity of the mercury and copper in R. caspicus and R. kutum effects of such hazardous pollutants so as to formulate the strategies for safeguarding aquatic organisms. Caspian roach (Rutilus caspicus) and Caspian kutum (Rutilus kutum) are found in the southern part of the Caspian Sea, particularly in the coastal waters of Iran, Turkmenistan and Azerbaijan. These species are economically important fishes (Coad, 2015). Hence, this study aimed to investigate the toxic effects of the mercury chloride and copper sulphate on the Caspian roach and Caspian kutum by determination of 96-hour LC50 values Materials and Methods The specimens of Caspian roach (n=120) and Caspian kutum (n=120), with mean weight of 3±0.6 g were obtained from Sijval restocking center (Bandar Turkaman, north of Iran) during August to September 2009. The fishes were transported in polythene bags to the fisheries laboratory of the Gorgan University of Agricultural Sciences and Natural Resources. Then, they were maintained in 20 L pre-cleaned glass aquaria filled with dechlorinated tap water with water temperature of 26±1°C in order to adaptation to the laboratory conditions. Thereafter, sets of 10 specimens (in triplicate) were randomly introduced to the aquarium systems. The exposure time to Hg (as HgCl2) and Cu (as CuSO4) were 96 hours, without any feed. Control group was designed with three replicates. No mortality was observed during the experimental period in control groups as well. In addition, four different concentrations of the mercury (as HgCl2) in geometric decreasing amounts of 0.02, 0.03, 0.04, and 0.05 mgL-1 were used. Also, four different concentrations of copper (as CuSO4) in geometric decreasing amounts of 1, 4, 7, and 10 mg L-1 in three- replicates were designed. For determination of the mortality limits of the mercury and copper as well as survival rate, the treatments and replications were considered based on OECD (OECD, 1988). Stock solutions (1000 mg L-1) were prepared by dissolving analytical-grade of the mercury (as HgCl2, Merck) and copper (as CuSO4; Merck) in distilled water. Preliminary tests were carried out to estimate the minimum lethal and maximum non- lethal concentrations of the mercury (as HgCl2) and copper (as CuSO4). Dissolved Oxygen (DO) (mg L - 1), temperature (°C), total hardness (mg L-1) and pH were recorded in each aquarium during experiment. The recorded water parameters were as: temperature=26±1°C, pH=7.7±0.2; total hardness=205 mg L-1 as CaCO3, and DO=7 mg L -1. Water quality of the experimental tank was determined according to standard procedures. Lethal concentration for 50% (LC50) values were calculated using the EPA computer probit analysis program (Version 1.5) (Vieira, 2009; Das et al., 2005; Pyle et al., 2002). Statistical analyses were performed using SPSS software (ver. 16.0, SPSS Co., Chicago, IL, USA). All the data were tested for normality using Kolmogorov-Smirnov test. Results The LC50 value for HgCl2 and CuSO4 in the Caspian roach and Caspian kutum at 24, 48, 72, and 96-hrs of exposure period were presented in Tables 1 and 2. The LC50 value in Caspian roach for HgCl2 at 24, 48, 72 and 96-hrs of exposure period were 0.64, 0.61, Metal LC50 values (mg L -1) 24 hours 48 hours 72 hours 96 hours Copper LC10 0.69 0.56 0.48 0.43 LC50 5.31 4.17 3.20 2.25 LC90 40.07 30.11 21.04 11.36 Table 1. Lethal concentration (LC50) of mercury and copper estimated by EPA method on the Caspian Roach (Rutilus caspicus). 27 Int. J. Aquat. Biol. (2016) 4(1): 25-30 0.42, and 0.28 mg L-1, respectively; while for CuSO4, the LC50 values at 24, 48, 72 and 96-hrs were 11.55, 5.08, 2.49, and 1.47 mg L-1, respectively (Table 1). Discussion The 96-hrs LC50 values of fish is species and metal dependent. In this study, the 96-hrs LC50 for HgCl2 was determined to be 0.28 mg L-1. In other studies the acute toxicity thresholds for inorganic mercury (typically as HgCl2) in freshwater organisms vary from approximately 0.005-0.230 mg L-1 in crustaceans, to 0.06-0.8 mg/L in fish (Ramamoorthy and Baddaloo, 1995). The 96-hrs LC50 values for freshwater fish ranges 0.033-0.4 mg L-1, while the LC50 is generally higher for marine fish (Boening, 2000). The 96-hrs LC50 values of HgCl2 on Capoeta fusca was 0.154 mg L-1 (Mansouri and Baramaki, 2011), on Gambusia holbrooki was 0.36 mg L-1 (Ebrahimpour et al., 2010a, b; Pourkhabbaz et al., 2011), and on Acanthoparus latus was 0.648 mg L-1 (Hedayati and Safahieh., 2010). In addition, the 96- hrs LC50 values of HgCl2 on Heteropneustes fossilis, Oncorhynchus mykiss, Roccus saxatilis, and Salvelinus fontinalis were found to be 0.35, 0.22, 0.09, and 0.075 mg L-1, respectively (Pandey et al., 2005). The acute toxicity of the copper sulphate decreased with increasing exposure time in both species. Copper sulphate was significantly (96-hrs LC50 values) more toxic on R. caspicus than R. kutum. The observed differences in the acting copper might be species dependent and their susceptibility rates to the test chemical, which resulted in their subsequent toxicity values. Copper salts combine with proteins present in the mucus of the fish's mouth, gills, and skin, preventing respiration causing death (Richey and Roseboom 1987; Peres et al., 1991). As conclusion, the mercury chloride was more toxic than copper sulphate on the Caspian Roach. In general, the toxicity of HgCl2 was higher than that of CuSO4 in both studied species. Acknowledgements The authors wish to express their gratitude to many people who have devoted their time and expertise to this project. We are also grateful to R. Tahergoorabi and S.V. Hosseini for their assistance in the preparation of the manuscript. References Abdullah S., Javed M. (2006). Studies on acute toxicity of metals to the fish, Catla catla. Pakistan Journal of Biological Sciences, 9: 1807-1811. Allen P. (1994). 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(2016) 4(1): 25-30 E-ISSN: 2322-5270; P-ISSN: 2383-0956 Journal homepage: www.ij-aquaticbiology.com © 2016 Iranian Society of Ichthyology چکیده فارسی Rutilus caspicus مس برروی کلمه ترکمنی سنجی سمیت حاد کلرید جیوه و سولفاتزیست Rutilus kutumماهی سفید، و 5ربابه واجدی، *1رجایی، قاسم 4، محمدحسین سینکاکریمی3،2برهان منصوری ،1علیرضا پورخباز .ایران ،بیرجند ،بیرجند دانشگاه طبیعی، منابع دانشکده ،علوم محیط زیستی گروه1 .ایران ،سنندج ،علوم پزشکی کردستان دانشگاه ،کمیته تحقیقات دانشجویی2 .ایران سنندج، ،علوم پزشکی کردستان دانشگاهمرکز تحقیقات بهداشت محیط، 3 .ایران ،مالیر ،مالیر دانشگاه ،و محیط زیست طبیعی منابع دانشکده ،علوم محیط زیستی گروه4 .ایران ،تهران ،تهران دانشگاه ،محیط زیست دانشکده ،علوم محیط زیستی گروه5 چکیده: سفید ماهی و( Rutilus caspicus) کلمه ترکمنی ماهی در مس سولفات و جیوه کلرید( 50LC) حاد سمیت تعیین حاضر مطالعه از هدف (Rutilus kutum )50 میزان. بودLC و ،42/0 ،91/0 ،94/0 ترتیببه مواجهه ساعت 69 و ،22 ،44 ،24 در جیوه کلرید برای ترکمنی کلمه ماهی ماهی 50LC مقادیر همچنین. آمد بدست لیتر در گرممیلی 42/1 و 46/2 ،04/5 ،55/11 ترتیب به نیز مس سولفات برای و بودند گرممیلی 24/0 مطالعه این از حاصل نتایج. بود لیتر درم گرمیلی 25/2 و 20/3 ،12/4 ،31/5 ترتیببه مواجهه ساعت 69 و ،22 ،44 ،24 در مس سولفات برای سفید ترینباال و یافت، کاهش زمان روند با میر ومرگ میزان. است بوده مطالعه مورد هایگونه برای مس سولفات از باالتر جیوه کلرید سمیت که داد نشان .داد رخ اول ساعت 24 طول در میر و مرگ میزان .ماهی استاتیک، سنجی زیست ،50LC سمیت، :کلمات کلیدی