Determination of Organochlorine Pesticide Residues in Water, Sediments and Fish from Lake Parishan, Iran

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World Journal of Fish and Marine Sciences 4 (2): 50-54, 202 ISSN 2078-4589 IDOSI Publications, 202 DOI: 0.5829/idosi.wjfms.202.04.02.56399 Determination of Organochlorine Pesticide Residues in Water, Sediments and Fish from Lake Parishan, Iran Farshid Kafilzadeh, Amir Houshang Shiva, 2 Rokhsareh Malekpour and Hamid Noorani Azad Department of Biology, Jahrom Branch, Islamic Azad University, Jahrom, Iran 2 Department of Biology, Kazeroun Branch, Islamic Azad University, Kazeroun, Iran Abstract: In this study, the levels of organochlorine (OC) pesticide residues in Lake Parishan have been investigated using water, sediment and fish (Barbus brachycephalus caspius) samples as a case study to find out the extent of pesticide contamination and accumulation in the lake. Six OC pesticides namely DDT, DDE, Lindane, Endosulfan, Heptachlor and Chlordane were analyzed in four sites at four seasons. Water samples were processed using a liquid-liquid extraction technique and gas chromatograph equipped with electron capture detector (GC-ECD). Soxhlet extraction was used for fish and sediment samples followed by clean up and gas chromatograph. DDE was the predominant residue in all the samples analyzed, at the mean concentrations of 0.055 ppb, 9.84 ppb and 4.86 ppb in water, sediment and fish samples, respectively. The lowest levels of OC pesticides were related to Heptachlor and Chlordane which none of them were found in water samples. Pol-e-Abgineh and Midstream sites had the highest and the lowest concentrations of OC pesticides, respectively. Key words: Organochlorine residues % DDT % Soxhlet % GC-ECD % Lake Parishan INTRODUCTION problem of chronic toxicity to animals and humans via air, water and foods intake. Many of these OC Organochlorine (OC) pesticides are among the pesticides and their metabolites have been implicated in agrochemicals that have been used extensively for long a wide range of adverse human and environmental effects periods. They have been used widely in agriculture, as including reproduction and birth defects [0], immune well as, in mosquito, termite and tsetse fly control system dysfunction, endocrine disruptions and cancer programs []. OC pesticides are characterized by low []. polarity, low aqueous solubility and high lipid solubility Fish are used extensively for environmental (lipophilicity) and as a result they have a potential for monitoring [2], because they uptake contaminants bioaccumulation in the food chain posing a great threat directly from water and diet. Generally the ability to human health and the environment globally [2]. of fish to metabolize organochlorines is moderate; Residues and metabolites of many OC pesticides are very therefore, contaminant loading in fish is well stable, with long half lives in the environment [3]. Studies reflective of the state of pollution in surrounding have shown that DDT is still in its highest concentration environments []. in biota of some areas. It is a hydrophobic molecule which Parishan (Famour) wetland [29 30' N, 5 47' E, alt. + + disrupts ionic channels like Na -K pumps in nervous cell 820 m] is located in 2 Km Southeast of Kazeroun, membrane leading to automatic stimulation of neurons Southern Iran. It has around.6-2 m depth and the surface and involuntary contraction of muscles [4]. 2 area is 0.8 Km. This lake is the only freshwater source in Many other recent works have indicated the Iran with rare species of birds and aquatic organisms. presence of OC residues in surface waters, sediments, Also it is one of the most important natural habitats for biota and vegetations [2, 5-9]. The persistent nature of migrating birds which migrate from Siberia, Canada, organochlorine residues in the environment may pose the Denmark and North Africa to Iran [3]. Corresponding Author: Farshid Kafilzadeh, Department of Biology, Jahrom Branch, Islamic Azad University, Jahrom, Iran. 50

The determination of OC residues in fish, sediments and water may give indication of the extent of aquatic contamination and accumulation characteristics of these compounds in the tropical aquatic biota that will help in understanding the behaviour and fate of these persistent chemicals [4]. This work, therefore, seeks to provide baseline information on levels of pesticide residues including DDT, DDE, Lindane, Endosulfan, Heptachlor and Chlordane in fish (Barbus brachycephalus caspius), sediments and surface waters of Parishan wetland through four seasons that will assist in a scientific assessment of the impact of pesticides on public health, agriculture and the environment in Iran. MATERIALS AND METHODS Fish, sediment and water samples were obtained from four various sites namely Gadarzavali, Hellak, Pol-e- Abgineh and Midstream in Lake Parishan. Samplings were conducted seasonally from winter 200 to autumn 200 following US-EPA [5]. Samples were collected from surface parts of the water and sediment. Also, each sampling was carried out in three replicates. A total of 48 samples each of sediments and water were collected randomly. However, fish samples were 4 because of little rainfall in recent years which has caused a decrease in water depth leading to limited dispersal of fish species. All samples collected (water, sediments and fish) were immediately stored in an ice-chest at 4 C and transported to the laboratory for analysis. Extraction of OC Pesticides in Water Samples: In the laboratory, using liquid-liquid extraction (LLE) as described in APHA [6], the total amount of each surface water sample (800 ml) was filtered with Whatman filter paper (i.d. 70 mm) to remove debris and suspended materials and then poured into a 2 liter separatory funnel. For the first LLE, the mixture of 00 ml n-hexane and dichloromethane (: v/v) was added and shaken vigorously for 2 min before two phase separation. The water-phase was drained from the separatory funnel into a 000 ml beaker. The organic-phase was carefully poured into a glass funnel containing 20 g of anhydrous sodium sulfate through a 200 ml concentrator tube. Following the second and third LLE, the water-phase was poured back into the separatory funnel to re-extract with 50 ml of the same solvent mixture. The extract was concentrated to the volume of 2 ml under a gentle stream of nitrogen using rotary evaporator and then analyzed with Gas Chromatography with micro Electron Capture Detector (GC-ìECD) [7]. Extraction of OC Pesticides in Fish and Sediment Samples: The muscle tissues of the fish samples were ground in a blender to obtain a homogenous composite, while the sediments were air-dried. OC residues in sediments and fish samples were extracted using Soxhlet Extraction [8]. A 0 g sample was placed into a beaker containing 50 g anhydrous sodium sulfate and mixed thoroughly. The sample mixture was transferred to an extraction thimble and placed in a Soxhlet extractor. The mixture was extracted with 50 ml of acetone: n-hexane (20:80 v/v) at 50 C for 4 h. The extracts were filtered, concentrated to ml using vacuum rotary evaporator. Each of the raw extracts was then dissolved in 0 ml hexane and passed through pre-conditioned octadecyl C-8 columns at a rate of 2 ml ming to clean up. The column was washed with ml, 30% methanol followed by ml ultrapure water and was allowed to dry. The sample (analyte) which was trapped in the column was eluted 5 times with 0.5 ml aliquots of hexane to recover the pesticide residues. Hexane in the sample was then allowed to evaporate off leaving the residue alone in the vial. Dried sample was dissolved in ml portion of hexane, mixed thoroughly with a whirl mixer and then transferred to auto sampler vials ready for gas chromatography [5]. Statistical analyses were carried out by analysis of variance (ANOVA) using SPSS 5 software. Mean values were analyzed by the Duncan s test. RESULTS AND DISCUSSION Parishan wetland is surrounded by farm lands. A large amount of chemicals (fertilizers and pesticides) are used there by farmers which can enter the wetland through running waters and subterranean canals. Also, garbage and wastewaters are poured in the wetland by inhabitants. All of these factors may lead to the contamination of Parishan Lake. Results from the study have been shown in tables, 2 and 3 which are related to the concentration of OC residues in water, sediment and fish samples, respectively. DDT was detected in 54% of water samples, 75% of sediments and 84% of fish samples. The associated figures for mean concentrations were 0.06 ppb, 4.980 ppb and 4.2 ppb. The ratios of incidence as well as the concentrations of DDE, a metabolite of DDT, in all the three sample types were higher than those recorded for DDT which was 84%, 96%, 94% in water, sediment and fish samples, respectively. This observed trend could be attributed to the decomposition and bioaccumulation of the DDT used in the past. DDE is more stable than DDT and decomposes more slowly by micro-organisms, heat and ultraviolet rays [4]. 5

Table : Levels of organochlorine pesticide residues in water samples of Lake Parishan DDT 0.06 0.003 0.2-0.24 54 DDE 0.055 0.008 0.05-0.4 84 Chlordane - - - - Heptachlor - - - - Lindane 0.058 0.02 0.45 -.50 68 Endosulfan 0.046 0.009 0.30-0.46 60 Table 2: Levels of organochlorine pesticide residues in sediment samples of Lake Parishan DDT 4.980.66 4.2-6.8 75 DDE 9.840 2.85 2.27-30.43 96 Chlordane 0.074 0.03 0.28-0.42 24 Heptachlor 0.08 0.04 0.24-0.44 36 Lindane 6.920.8 3.06-4.2 72 Endosulfan 0.622.72 4.2-5.22 46 Table 3: Levels of organochlorine pesticide residues in fish muscle tissue samples of Lake Parishan DDT 4.2.2 3.50-5.56 84 DDE 4.864.42 4.8-8.26 94 Chlordane 0.024 0.008 0.35-0.56 2 Heptachlor 0.04 0.0 0.28-0.58 34 Lindane 0.38 0.06 0.65 -.47 39 Endosulfan 0.86 0.2 0.94-2.4 33 Neither chlordane nor heptachlor was detected in the concentration in water. It is therefore expected that water samples showing that the farmers around the lake lindane levels in fish will rise with time as they utilize the do not use them in their farming activities. However, these water and the sediments in the lake. organochlorine pesticides were measured in fish and Endosulfan, a broad spectrum contact insecticide sediment samples, which is because they are less soluble and acaricide, is another pesticide used by many farmers. in water. They, therefore, accumulate in fishes and The highest concentration of endosulfan (5.22 ppb) was sediments when they are discharged into water bodies. detected in the sediment samples, however, the highest Frequency of detection (incidence) and concentration of occurrence of detectable concentrations was found in chlordane and heptachlor measured in sediments were water (60%). Endosulfan level in fish, on the average, is 6 higher than those recorded for fish samples. Chlordane times that found in water samples. This suggests that the was detected in 24% of sediment samples at residues in the water are accumulated and concentrated in concentrations ranging from 0.28 to 0.42 ppb, while it was the fish. Mean level of endosulfan in the sediments is detected in 2% of the fish sample and their about 3 times the levels measured in fish. concentrations ranged between 0.35 and 0.56 ppb. Bioconcentration of endosulfan in fish could therefore Heptachlor was detected at an average concentration of arise. 0.08 ppb and 0.04 ppb in sediments and fish, In general, the concentration of OC pesticides in respectively. water was lower than sediments and fishes which was Lindane was detected in 68% of water samples, 72% highly significant (P?0.05), because these pesticides are of sediment samples and 39% of fish samples lipophilic and are not soluble in water. This fact can ease analyzed with the highest concentration of 4.2 ppb the accumulation of OC pesticides in micro-organism occurring in sediments. This result suggests lindane is tissues. more prevalent and persistent in the sediments than in The mean total concentration of organochlorine water and then in fish. The average concentration of residues in water samples of Gadarzavali, Hellak, lindane in sediments was about 50-folds the average Pol-e-Abgineh and Midstream sites were 0.023 ppb, concentration in fish and 20-folds the average 0.030 ppb, 0.054 ppb and 0.00 ppb, respectively (Fig. ). 52

Fig. : Comparison among the mean total concentration of organochlorine pesticides in water samples of four sites Fig. 2: Comparison among the mean total concentration of organochlorine pesticides in sediment samples of four sites Based on ANOVA and Duncan tests, the mean respectively. In terms of DDE, the respective figures were concentrations of OC pesticides in water samples of Pol-e- 0.06±0.03 nggg, 8.34±2.96 nggg and 5.23±.30 nggg. Abgineh and Midstream sites showed significant In this study, our results show that concentration of DDE differences (P?0.05). However, these differences were not is more than DDT. significant between Gadarzavali and Hellak sites (P?0.05). According to Siriwong et al. [7] the rate of DDT and In terms of the mean concentrations of OC residues in its derivatives in sediments was 2.05 nggg and in sediments, as fig. 2 illustrates, all four mentioned sites vertebrates was 4.6-57.66 nggg which were more than showed different levels which were highly significant the results obtained in this investigation. (P?0.05). Generally, the highest concentration of OC The levels of most of the residues in fish were higher pesticides was seen in Pol-e-Abgineh. It may be due to than those found in water. Organochlorine pesticide the abundance of farm lands around this site which have residues in the lake are likely to originate from nonpoint sharp slopes toward this part of the lake, so pesticides sources via runoff, atmospheric deposition and leaching and other chemical materials can enter there more easily. due to agricultural applications and vector control However, the lowest concentration of organochlorine practices. The lake sediments act as a sink for the residues was related to Midstream site. Because there is persistent contaminants, whose resuspension during the a slow current toward this part and the contaminants lake s mixing may increase pesticide bioavailability and cannot accumulate there. accumulation in the fish. Pesticide pollution to the lake is Hardell et al. [9] reported the rate of chlorine therefore, likely to pose a danger to both aquatic pesticides in muscle tissue of fishes and DDE was the organisms and humans. predominant target compound. The results obtained in this study generally agree with their findings. ACKNOWLEDGEMENTS The amount of OC pesticides in freshwater fishes of Punjab was measured in another study and The authors would like to thank the Office of Vice showed the predominance of DDT, while other Chancellor for Research of Islamic Azad Universityorganochlorine pesticides such as lindane, alderine, Jahrom branch for collaboration in some parts of this dielderine, chlordane, endosulfan and heptachlor were study. found at lower levels [20]. These levels reported in India were extremely high compared to levels recorded in this REFERENCES work. Some investigations have been done on OC residues. Guo, Y., X.Z. Meng, H.L. Tang and E.Y. Zeng, 2008. in fish (Tilapia), sediment and water samples from Lake Tissue distribution of organochlorine pesticides in Bosomtwi in Ghana by Darko et al. [5]. The mean fish collected from the Pearl River delta, China: concentrations of DDT in water, sediment and fish were Implications for fishery input source and 0.02±0.62 nggg, 4.4±.54 nggg and 3.64±.8 nggg, bioaccumulation. Environ. Pollut., 55: 50-56. 53

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