International Journal of Environmental Biology

Size: px

Start display at page:

Download "International Journal of Environmental Biology"

Garey Kelley
6 years ago
Views:

Available online at http://www.urpjournals.com International Journal of Environmental Biology Universal Research Publications.

calcarifer (Bloch, 1790). R. Ezhilmathy *, K. Raja Lakshmi, A. Chezhian, N. Kabilan, D. Senthamilselvan.

1 Available online at International Journal of Environmental Biology Universal Research Publications. All rights reserved ISSN X Original Article Evaluation of the joint action toxicity of mixtures of pesticide (Profenofos) and heavy metal (lead nitrate) against the estuarine fish Lates calcarifer (Bloch, 1790). R. Ezhilmathy *, K. Raja Lakshmi, A. Chezhian, N. Kabilan, D. Senthamilselvan. Centre of Advanced Study in Marine Biology, Annamalai University, Parangipettai, Tamil Nadu, India * Corresponding Author: R. Ezhilmathy, Research Scholar, CAS in Marine Biology, Annamalai University, Parangipettai Tamilnadu, India. Phone No ezhilbiotech86@gmail.com Received 15 November 2014; accepted 27 November 2014 Abstract The present study was carried out to evaluate the effects of pesticide and metal (Profenofos, lead nitrate) on the haematology of fish, Lates calcarifer for every 24 h. up to 96 h. The specimens were exposed to acute concentration of individual pesticide (Treatment II) and metal (Treatment III) and mixed pesticide and metal (Treatment IV) viz., Profenofos (30ppb), lead nitrate (2.8 ppm) and profenofos plus lead nitrate (18 ppb) for a period of 96 h. When the fishes were exposed to above all three treatments the haematological changes, like haemoglobin and haematocrit were changed drastically. Blood haemoglobin level was decreased in all the three treatments throughout the exposure period showing a minimum percentage decrease of -21.4, and at the end of 24 hours of treatments, respectively. Whereas the maximum percentage decrease of , and at the end of 96 hours of treatments, respectively. The Hematocrit level also decreased in all the three treatments throughout the exposure period showing a minimum percentage decrease of , and were observed at the end of 24 hours, respectively. The maximum percentage decrease of , and were observed in all the treatments at the end of 96 hours, respectively. The decrease in the haemoglobin and haematocrit level was more in the treatment-iv, when compared to treatments II, and III. From this investigation, the results of acute toxicity proved that the joint action of mixture of pesticide and metal toxicity induced haematological changes is more than individual pesticide and metal toxicity of fish, Lates calcarifer Universal Research Publications. All rights reserved Introduction: The rate of toxic substances released in the environment has increased rapidly due to various human activities (Chakkaravarthy, 2003). The advancement in technology and increased global industrialization has increased the production of toxic heavy metals e.g. lead, and the aquatic environments often serve as the sink for these substances (Sobha et al., 2007). In the recent years, there have been a noticeable increase in the usage of agricultural pesticides; this could be in response to increasing global population which have exerted more pressure on human to respond to the attendant problem of food insecurity arising from global population explosion (Pimentel, 2001). Investigations into the effects of pesticides and metals on fish have a diagnostic significance in evaluating the adverse effects of these toxicants to human health. Profenofos is one of the recommended insecticides and a preferred one over the conventionally employed toxic chemical pesticides (Venkateswara Rao, 2003). Profenofos is an organophosphorous insecticide used for various agricultural and household purposes to control insects and other pests (IPCS, 1990: U.S. EPA, 1997). Lead is a nonbiodegradable naturally occurring heavy metal and may potentially become accumulated in various tissues of aquatic organisms. There are reports on the toxicity of Pb in fish; alteration of biochemical and haematological parameters in the different fishes were reported (Ates et al., 2008; Maiti et al., 2010 and Martinez et al., 2004). Among the aquatic species, the fish are the major targets of toxicants contamination. Fish are largely being used for the assessment of the quality of aquatic environment and as such can serve as bioindicators of environmental pollution. Blood is a pathophysiological 248

2 reflector of the whole body, and therefore, blood parameters are important in diagnosing the structural and functional status of fish exposed to toxicants (Adhikari et al. 2004). Most studies on the effects of environmental pollutants are limited to reporting the effects of either pesticide treatment or metal exposure individually, and only few reports are available on the effects of multiple stressors of pesticides contamination and heavy metal pollution on aquatic organisms. Studying the effects of mixture of contaminants on organisms is particularly necessary because aquatic organisms are exposed to combinations of stressors in their environment. The work on combined toxicity of organophosphorous pesticide (Profenofos) and heavy metal (lead nitrate) is very scanty; hence the present study was designed to determine the toxic impact of pesticide plus metal toxicity on the haematology of the fish Lates calcarifer. Materials and methods: Specimens of L. calcarifer were procured from Rajiv Gandhi Centre for Aquaculture (RGCA), Thirumullaivasal, Sirkali, Tamil Nadu- India and acclimatized to the laboratory conditions for 15 days. Water was changed daily and fish were fed ad-libitum with a mixture of rice bran, groundnut oil cake and dry fish, twice a day. Fish ranging from 7-8 cm in length and weighing 8-10g were selected for experimental purpose. The quality of the water was determined according to APHA et al., 1976 were as follows; Dissolved oxygen 5.59 mg/l ; ph 7.9± 0.2; Water temperature- 35± 2ºC; Salinity- 29±0.07ppt; Total hardness- 8.0±0.2 mg/l; Magnesium- 3.0±2.0 and Total alkalinity - 16±.06mg/l. The fishes were introduced into glass aquarium (75 x 35 x 75 cm) of 150 L capacity which was washed thoroughly. Fish belonging to both the sexes were used. Preliminary studies were carried out to find out the median lethal concentration (LC 50) for 96 hrs of Pesticide (Treatment-II), metal (Treatment- III) and Pesticide plus metal (Treatment IV). For this, appropriate amount of Pesticide, metal and Pesticide plus metal were dissolved in seawater freshly every time to prepare a stock solution of 1000 ppm for each toxicant. Then different wide and narrow range trials of toxicity test were conducted based on the survival/ mortality of the experiment. The median lethal concentration (LC 50) of fish for 96 hrs Pesticide (30ppb), metal (2.8 ppm) and Pesticide plus metal (18 ppb) were determined and calculated by probity analysis method of (Finney, 1978 ). For acute studies, four plastic tub of 10 L capacity and each with 10 L of water were taken and to the first three tubs, 96 hrs LC 50 concentrations of each toxicant were added respectively, while the fourth tub served as a control (Treatment- I). Then, 10 fishes were introduced in each tank and the experiments were maintained for a period of 96 hrs. Fish were fed adlibitum, and water and toxicants were renewed daily. The mortality/survival time of fish in each tub was observed for 24h. No mortality was observed in the control. The concentration at which the 50% kill of fish occurred after 24h treatment was taken as median lethal concentration (LC 50) for 24h. At the end of every 24 h, live fishes from each experimental tank were taken and blood was drawn from the heart region by cardiac puncture, with heparin as an anticoagulant and transferred into small vials kept in ice-cold condition. From the collected blood samples the haemoglobin content and hematocrit level were estimated. The haemoglobin content was estimated by the cyanmethemoglobin method using diagnostic reagent kit and hematocrit or packed cell volume were analysed by microhematocrit (capillary) method. Student s t test was used to study the significance. Results: Table 1 and 2 shows the changes in the haemoglobin (Hb) content and haematocrit (Hct) levels of fish L. calcarifer exposed to Pesticide, metal and Pesticide plus metal concentration for a period of 96h. of treatment. In all the three treatments the hemoglobin content was decreased throughout the exposure period showing minimum percent decrease of -21.4, and at the end of 24 h, respectively. Whereas the maximum percent decrease of , and were observed at the end of 96 hours of exposure, respectively. Similarly, the hematocrit level was also decreased throughout the exposure period showing minimum percent decrease of , and at the end of 24 hours, respectively and the maximum percent decrease of , and at the end of 96 h. of treatments, respectively. The per cent decrease was more in treatment- IV, when compared to that of the treatment- II, III. Discussion: The haematological picture is frequently utilized for the detection of pathophysiological changes in different stress conditions such as exposure of toxicants (Nussey et al., 1995). In the present study, in all three acute treatments the decreased level of haemoglobin and haematocrit was observed, similar observation was made by Masoud et al., (2012); Rostami et al., (2005) in Pesticide (diazinon toxicity) exposed to rainbow trout fry Oncorhynchus mykiss, and Venkataraman and Sandhya Rani (2013) reported that decrease in the concentration of hemoglobin in blood of fish Clarias batrachus exposed to malathion, he further stated that a decrease in the haematocrit indicates the worsening of an organism state and developing anemia. Magar and Dube, (2012) reported that the significant decrease in haemoglobin content in malathion exposed fish Channa punctatus may be due to release of immature cells from haemopoietic tissue into the blood strength as well as disruption of iron metabolism that lead to a defective haemoglobin synthesis. Omoregie et al. (1994) also absorbed decreased level of haemoglobin in Oreochromis niloticus was exposed to formalin. He further reported that the significant decrease in the haemoglobin concentrations may also be due to a decrease in the rate of haemoglobin synthesis (Reddy and Bashanihideen, 1989). Adakole (2012) showed significant decrease in haematological indices in metal finishing company effluent exposed fish Clarias gariepinus fingerlings. The author further reported that alteration in the blood may be due to alter in the properties of hemoglobin by decreasing their affinity towards oxygen binding capacity rendering the erythrocytes more fragile and permeable which probably results in cell swelling 249

3 Table-1. Changes in haemoglobin in the blood of L. Calcarifer exposed to acute concentration of pesticide, metal and pesticide plus metal. Exposure period (in hours) Control Treatment- I ± ± ± ±0.052 Blood haemoglobin content (g%) Treatment-II T-test Treatment-III T-test Treatment-IV T-test 5.137±0.056 (-21.4 ) 4.582±0.036 ( ) 4.171±0.024 ( ) 3.386±0.069 ( ) ±0.007 ( ) 4.821±0.018 ( ) 4.338±0.013 ( ) 4.068±0.009 ( ) ±0.004 ( ) 4.082±0.015 ( ) 3.976±0.005 ( ) 3.516±0.013 ( ) Table-2. Changes in haematocrit in the blood of L. Calcarifer exposed to acute concentration of pesticide, metal and pesticide plus metal Exposure period (in hours) Control Treatment-I ± ± ± ±0.078 Blood haematocrit content (g%) Treatment-II T-test Treatment-III T-test Treatment-IV T-test ±0.027 ( ) ±0.087 ( ) ±0.032 ( ) ±0.037 ( ) ±0.027 ( -9.48) ±0.038 ( ) ±0.047 (-23.96) ±0.057 ( ) ±0.087 (-19.02) ±0.032 ( ) ±0.021 ( ) ±0.056 (-41.80) deformation and damage. Nussey et al., (1995) reported that the decreased in haemoglobin concentration of tilapia, Oreochromis mossambicus exposed to copper signifies that the fish ability to provide sufficient oxygen to the tissues is restricted considerably and will result in decrease of physical activity. Reduction of haematological indices have also been reported by Kori-siakpere and Ikomi (2011) in Parachanna Africans exposed to cadmium may be due to alterations in the haemolysis and impairment of haemoglobin synthesis, resulting in a hypochromic microcytic anaemia. Shah and Altindag, (2004) observed decrease in haemoglobin, Red blood cell (RBC) count and Hct in fish Tinca tinca exposed to mercuric chloride and lead and the author further reported that the reason may be due to the disturbed haemoglobin synthesis and this may result in anaemia. Vutkuru, (2005) and Shalaby, (2001) reported that a significant decrease in RBC s hemoglobin and packed cell volume of fish exposed to heavy metals and the reason behind this may be disturbed haemoglobin synthesis. The blood cell count (RBC and WBC) and hemoglobin (Hb) were significantly lower in endosulfan exposed fish Oreochromis mossambicus was observed by Neeraj Kumar et al., (2011). The author further reported that the reason behind the reduction in hematological values indicates anemia in the pesticide exposed fish, which may be due to erythropoietic, haemosynthetic and osmoregulatory dysfunction or due to an increase in the rate of erythrocyte destruction in haematopoietic organs (Jenkins et al., 2003; Seth et al., 2003). In the present study, the decrease levels of haemoglobin and heamatocrit may be due to anaemic effect or decrease in oxygen or increased destruction of hemoglobin or a decrease in the rate of haemoglobin synthesis. It should be noted that this mixture of pesticide (Carbamate) and heavy metal (Zinc) were determined in the blood of fish, Labeo boga showed decline in haemoglobin and heamatocrit may be attributed to RBC lysis/destruction and greater prevalence of erythroblasts (immature RBCs) in the general circulation (Raina and Sachar, 2014). Similarly in the present study, decreased level of haemoglobin and heamatocrit to individual and mixture of pesticide and metal exposed fish Lates calcarifer were also observed and this may be due to findings of above reasons. Conclusion In the present study an attempt was made to assess toxic impact of profenofos (Treatment I), lead nitrate (Treatment II), Profenofos plus lead nitrate (Treatment III) on haematology of fish, Lates calcarifer. In the present study, decreased level of haemoglobin and heamatocrit to individual and mixture of pesticide and metal was observed. The decrease in the haemoglobin and haematocrit level was more in the treatment-iv, when compared to treatments II, and III. This work underscores the need for new studies to explore how complex mixtures of pesticide and metal can affect physiological responses of aquatic organisms. Acknowledgement We thank the authorities of the Annamalai University for providing the necessary facilities. The first author thanks the DST PURSE Programme, Department of Science and Technology (Government of India) for the 250

4 financial support during the period of study. References 1. Adhikari S, Sarkar B, Chatterjee A, Mahapatra CT, Ayyappan S (2004) Effects of cypermethrin and carbofuran on certain hematological parameters and prediction of their recovery in a freshwater teleost, Labeo rohita (Hamilton). Ecotoxol Environ Saf 58: APHA., AWWA and WPCF., Standard methods for the examination of water and waste water 13 th (Ed.), American public health association, Washington, USA. 3. Ates B, Orun I, Talas ZS, Durmaz G, Yilmaz I (2008). Effects of sodium selenite on some biochemical and haematological parameters of rainbow trout (Oncorhynchus mykiss Walbaum, 1792) exposed to Pb2+ and Cu2+. Fish Physiol Biochem 34: Chakkaravarthy Q (2003). Human Survival and Environmental Pollution. In: Bunch MJ, Madha S, Kumaran TV Eds. Proceedings of the Third International Conference on Environment and Health, Chennai, India, December, Chennai: Department of Geography, University of Madras and Faculty of Environmental Studies, York University, p Finney DJ (1978). Statistical Methods in Biological Assay. 3rd Edn. Griffin Press, London. 508 p. 6. G.V. Venkataraman and Sandhya Rani (2013). Acute toxicity and blood profile of freshwater fish, Clarias batrachus (Linn.) exposed to Malathion.Journal of Academia and Industrial Research. Volume 2, Issue IPCS. (1990). The WHO recommended classification of pesticides by hazard and guidelines to classification. Geneva: World Health Organization, Environmental Health Criteria, 94: J. A. Adakole (2012). Changes in some haematological parameters of the African catfish (Clarias gariepinus) exposed to a metal finishing company effluent Indian Journal of Science and Technology Vol. 5 No. 4: Jenkins, F., Smith, J., Rajanna, B., Shameem, U., Umadevi, K., Sandhya, V., Madhavi, R., Effect of sublethal concentrations of endosulfan on haematological and serum biochemical parameters in the carp Cyprinus carpio. Bull. Environ. Contam. Toxicol. 70, Khoshbavr Rostami, H., A.S. Soltani, M. Muhyiddin and H. Aldyt the Haj, Changes in some hematological and biochemical parameters in blood serum of Beluga (Huso huso) after prolonged exposure to the pesticide Diazinon. Iranian Fisheries Science (English) 5: 66 to Larsson A, Haux C, Sjobeck M (1985). Fish physiology and metal pollution. Results and experiences from laboratory and field studies. Ecotoxicol Environ Safety 9: Magar R.S and Dube K.V (2012). Impact of malathion on some haematological parameters of channa punctatus (bloch). IJBAR, 03(09). 13. Maiti AK, Saha NC, Paul G (2010). Effect of lead on oxidative stress, Na+ K+ATPase activity and mitochondrial electron transport chain activity of the brain of Clarias batrachus L. Bull Environ Contam Toxicol 84: Martinez CBR, Nagae MY, Zaia CTBV, Zaia DAM (2004) Acute morphological and physiological effects of lead in the Neotropical fish, Prochidolus lineatus. Braz J Biol 64: Masoud Saeedi Far, 1Habib Vahabzadeh Roodsari, Abasali Zamini, 2Elham Mirrasooli and Rizvan Kazemi (2012). The Effects of Diazinon on Behavior and Some Hematological Parameters of Fry Rainbow Trout (Oncorhynchus mykiss) World Journal of Fish and Marine Sciences 4 (4): N. Seth, K.K. Saxena, Hematological responses in a freshwater fish Channa punctatus due to fenvalerate Bull, Environ. Contami. Toxicol. 71 (2003) Neeraj Kumar, P. Antony Jesu Prabhu, A.K. Pal, S. Remya, Md. Aklakur, R.S. Rana, Subodh Gupta, R.P. Raman, S.B. Jadhao (2011). Anti-oxidative and immuno-hematological status of Tilapia (Oreochromis mossambicus) during acute toxicity test of endosulfan. Pesticide Biochemistry and Physiology 99: Nussey G, Van Vuren JHJ, Du Preez HH (1995). Effects of copper on haematology and osmoregulation of the Mozambique tilapia, Oreochromis mossambicus (Cichlidae). Comp. Biochem. Physiol.111: Omoregie E, Eseyin TG, Ofojekwu PC (1994). Chronic effects of formalin on erythrocyte counts and plasma glucose of the Nile Tilapia Oreochromis niloticus. As Fish Sci 7: Ovie KORI-SIAKPERE, Urowoli IKOMI (2011). Alterations in Some Haematological Parameters of The African Snakehead: Parachanna africans Exposed to Cadmium. Not Sci Biol, 2011, 3(4): Pimentel D (2001). Overview of the use of genetically modified organisms and pesticides in agriculture. Ind J Global Legal Stud 9: Reddy PM, Bashamohideen M (1989). Fenvalerate and cypermethrin induced changes in the haematological parameters of Cyprinus carpio. Acta Hydrochem. Hydrobiol. 17: Shah SL, Altindg A (2004). Hematological parameters of tench (Tinca tinca L) after acute and chronic exposure of lethal and sublethal mercury treatments. Bull. Environ. Contamin. Toxicol. 73: Shalaby AM (2001) Protective effect of Ascorbic acid against Mercury intoxication in Nile tilapia (Oreochromis niloticus). J. Egypt. Acad. Soc. Environ. Develop. (D- Environmental studies). 2(3), Sheetu Raina, Anupriya Sachar (2014). Effect of Heavy Metal, Zinc and Carbamate Pesticide, Sevin on Haematological Parameters of Fish, Labeo Boga. International Journal of Innovative Research in Science, Engineering and Technology Vol. 3, Issue Sobha K, Poornima A, Harini P, Veeraiah K (2007). A study on the biochemical changes in freshwater fish Catla catla exposed to the heavy metal toxicant, cadmium chloride. Kathmandu University J 251

5 Sci Engineer Technol 3: U.S. EPA. (1997). Department of pesticides regulation, medical toxicology branch: summary of toxicology data Profenofos. pp Washington, DC: U.S. EPA. 28. Venkateswara Rao J, Shilpanjali D, Kavitha P, Madhavendra SS Toxic effects of profenofos on tissue acetylcholinesterase and gill morphology in a euryhaline fish, Oreochromis mossambicus. Arch Toxicol 77: Vutkuru SS (2005) Acute effects of Hexavalent chromium on survival, oxygen consumption, Hematological parameters and some biochemical profiles of the Indian Major Carp, Labeo rohita. Int. J. Environ. Res. Public. Health. 2, Source of support: Nil; Conflict of interest: None declared 252

Haematological and Serological Responses of Clarias Gariepinus to Sublethal Concentrations of Lead Nitrate

Haematological and Serological Responses of Clarias Gariepinus to Sublethal Concentrations of Lead Nitrate ABSTRACT: C.F. Ikeogu 1, C.I. Nsofor 2, I.O. Igwilo 3, A.A. Ngene 4 1Department of Fisheries and