To evaluate the quality and quantity of pollution and their possible damage to aquatic life, water and fish samples were collected from different locations along the River Kabul. Water samples both from upstream and downstream from polluted portion of River Kabul, both during high flow and low flow seasons, and effluent samples from drains of two major factories viz. Textile and Paper Industries and Ferozsons Laboratories and Associated Ghee Industries Limited. at Aman Garh Industrial Area. Nowshera, were analyzed for various physical and chemical properties and heavy metal content and compared with those of water samples from Warsak Dam (control) and National Environmental Quality Standards (NEQS).
Materials and Methods 1 Water samples for heavy metal estimations were collected in separate 1 litre plastic bottles and were preserved with 5 ml IINO3 (55%) per litre to prevent metal adsorption on the inner surface of the container. Samples for sulphide were separately collected and their sulphide content was fixed with 1.5 ml zinc acetate solution per liter sample on the spot. All water samples were then stored at 4°C in refrigerator for chemical and physical analyses.
2. The water samples were analyzed for temperature. appearance. odor. pH. dissolved oxygen (DO), conductivity. organic matter in the sediment. total suspended solids (TSS) and total dissolved solids (TDS). The chemical parameters included biological oxygen demand (BOD). chemical oxygen demand (COD). total hardness as CaCO3. calcium hardness as CaCO3. magnesium hardness as CaCO3, alkalinity as CaCO3, sodium. potassium. NH3, nitrite. nitrate. chloride. sulfate. phosphate. and sulfide contents. Heavy metals like chromium. Zine copper. nickel and lead were also determined in the water samples.
3 Fish samples were collected from different areas of highly polluted part of the man river. One fish sample was collected from the part of river receiving industrial waste (Site 1), whereas the second fish sample was taken from the region, where Nowshera city sewage and dirty Kalpani Canal (bringing sewage from Mardan, Risalpur and other adjacent towns) also join River Kabul (Site 2). Both the above samples were considered as fish samples from polluted water (treated fish samples) and were compared with the third fish sample collected from almost the non polluted Warsak Dam, about 60 km upstream the polluted part of the River Kabul (Site 3).
4. The fish samples, from control as well as polluted waters, were analyzed and compared for various hematological, blood, muscle and liver biochemical parameters and for heavy metals to assess the effect of water pollution on the fish health with extrapolation to hazards to human health.
5. The body weight, body length, standard length, total length, fork length, post operculum length, head length, eye diameter, mouth gape, length of lower jaw, length of upper jaw, and lengths of fins like pelvic, pectoral, dorsal. anal and caudal, of the fish were recorded. After morphometric measurement the fish were dissected, livers, portions of muscle, skin and gills were taken out. washed with distilled water, and stored in freezer (at -20°C) for further analyses.
6. Blood samples collected in tubes having ethylenediaminetetraacetic acid (EDT A) were used for hematological studies, and for the estimation of various biochemical components. Hematological parameters included hemoglobin (Hb), packed cell volume (PCV), red blood cell (RBC) count, total leukocyte count (TLC), mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC). Plasma was analyzed for various biochemical parameters like total protein, cholesterol, glucose, glutamate oxaloacetate transaminse (GOT), glutamate pyruvate transaminase (GPT) and creatine phosphokinase (CPK).
7 Liver was processed and analyzed for various biochemical parameters like total protein, soluble protein. total cholesterol. total lipid, glucose. free amino acid, DNA. RNA. amylase. GOT, GPT and lactate dehydrogenase (LDH) contents. 8 White muscle was processed and analyzed for various biochemical parameters like total protein, soluble protein., total cholesterol. total lipid, glucose, free amino acid, DNA. RNA, free fatty acids, amylase, GOT, GPT and LDH contents.
9 Gills, liver, muscle and skin tissues were processed and analyzed for the estimation of heavy metals like chromium, copper, nickel, lead and zinc.
Results IThe effects of wastewater pollution on river water quality are summarized as under: 1 Water sample A from Warsak Dam water reservoir had temperature, pH, DO and electrical conductivity in the range of 19-21oC, 7.0-8.0, 6.6-8.9 mgO2/1 and 166-260 μS/cm, respectively, while SOM, TSS and TDS range was 0.9-2.0%, 380-600 mg/1 and 150-1220 mg/1, respectively. Different chemical parameters at this site were; BOD 0.4-0.9 mgO2/1, COD 19-23 mgO2/1, free CO2, 1.1-2.34 mg/1, total hardness as CaCO3 79-167 mg/1, calcium hardness as CaCO3., 55-100 mg/1. magnesium hardness as CaCO3 20-69 mg/1, total alkalinity as CaCO3 50-85 mg/1. sodium 2-10 mg/1, potassium 0.5-2.1 mg/I, ammonia 0.002-0.02 mg/1. nitrate 0.05-1.49 mg/1, nitrite was negligible throughout the study at this location, chloride 5-8 mg/1, sulphate 15-28 mg/1. phosphate 0.001-0.16 mg/1 and sulphide 0.09-0.21 mg/1. Among the heavy metals, chromium ranged between 0.002-0.12 mg/1, zinc 0.011-0.091 mg/1. copper 0.01-0.091 mg/1. nickel 0.001-0.03 mg/1. and lead 0.002-0.150 mg/1.
2. Water sample B from River Kabul upstream or the confluence point where industrial wastes joined the river had temperature 18-27°C, pH 7.1-8.2, DO 5.8-6.9 mgO2/1 and electrical conductivity 220-400 μS/cm, respectively. while SOM, TSS and TDS occurred in the range of 2.2-4.85%, 210-900 mg/1 and 250-615 mg/1. respectively. Sample B had different chemical parameters in the ranges; BOD 1.60-3.9 mg 02/1, COD 34-48 mg02/1, free C02 3.2-5.2 mg/1, total hardness as CaCO3 90-171. 9 mg/1, calcium hardness as CaCO3 50-110 mg/1, magnesium hardness as CaCO3 40-71 mg/1, total alkalinity as CaCO3 50-110 mg/1, sodium 13-28 mg/1, potassium 2.1-3.9 mg/1, ammonia 0.14-0.526 mg/1, nitrate 1.00-1.99 mg/1, nitrite 0.009-0.021 mg/1, chloride 13-22 mg/1, sulphate 30-80 mg/1, phosphate 0.03-0.24 mg/1 and sulphide 0.23-0.74 mg/1. Among the heavy metals the water sample 8 had 0.048-0.263 mg/1 chromium, 0.172-0.4211 mg/1 zinc. 0.24-0.391 mg/1 copper, 0.01-0.09 mg/1 nickel and 0.02-0.04 mg/l lead.
3 Sample C representing the combined effluents from Textile and Paper Industries had temperature 25°C, pH 8.1-8.6, DO 0.3-0.8 mgO2/1 and electrical conductivity 707-796 μS/em, while SOM, TSS and TDS occurred in the range of 20.2-29.3%. 470-620 mg/1 and 645-810 mg/1, respectively. The water sample C had BOD 275-304 mgO2/1, COD 400-636 mgO2/1, free CO2 25.6-41.1 mg/1, total hardness as CaCO3 400-500.7 mg/1, calcium hardness as CaC03 280-425 mg/l, magnesium hardness as CaC03 71-210 mg/1, total alkalinity as CaCO3 224-401 mg/1, sodium 94-115 mg/1, potassium 15-19 mg/1, ammonia 2.3-3.7 mg/1, nitrate 0.4-1.5 mg/1, nitrite 0.1-0.2 mg/1, 29-40 mg/1, sulphate 395-621 mg/1, phosphate 1.39-2.71 mg/1 and sulphide 496-575 mg/1. The water sample Chad 0.41-0.71 mg/1 chromium, 0.06-0.92 mg/1 zinc, 0.06-0.17 mg/1 copper, 0.01-0.03 mg/1 nickel and 0.045-0.15 mg/1 lead.
5. The water Sample E. taken downstream from the confluence point from River Kabul. had 18-29 oC temperature, 7.1-7.6 pH, 4.8-5.8 mgO2/1 DO and 200-450 μS/cm electrical conductivity. while SOM, TSS and TDS at this point occurred in the range of 8.2-13.3%. 397-995 mg/1 and 225-690 mg/1. respectively. Water sample E had BOD 3.7-5.9 mgO2/1. COD (1)-178 mgO2/1. free C02 5.5-6.9 mg/1.tlolal hardness as CaCO3 95-190 mg/1. calcium hardness as CaCO3 56-130 mg/1. magnesium hardness as CaCO3 35-80 mg/1. total alkalinity as CaCO3 100-271 mg/1. sodium 10-34 mg/1. potassium 3.1-6.5 mg/1. ammonia 0.21-0.671 mg/l. nitrate 1.01-1.97 mg/l. nitrite 0.03-0.09 mg/l. chloride 15-27.1 mg/1. sulfate 71-193.7 mg/1. phosphate 0.034-0.261 mg/1. and sulphide 0.26-0.77 mg/1. This water sample had 0.01-0.31 mg/1 chromium. 0.029-0.49 mg/1 zinc. 0.32-0.49 mg/1 copper. 0.021-0.071 mg/1 nickel and 0.006-0. 121 mg/1 lead.
6. Water sample F from River Kabul downstream to sampling point E had temperature 19-28oC. pH 7.1-7.9. DO 4.8-8.2 mgO2/1 and electrical conductivity 185-440 μS/cm. while SOM, TSS and TDS occurred in the range of 8.9-13.7%. 400-890 mg/1 and 275-710 mg/1. respectively. Sample F had BOD 5.0-7.4 mgO2/1. COD 77-185 mgO2/1. free C026.8-7.9 mg/. total hardness as CaCO3 97-201 mg/1. calcium hardness as CaCO3, 55- 132 mg/1. magnesium hardness as CaCO3 38-80 mg/1. total alkalinity as CaCO3-101-210 mg/1, sodium 12-34 mg/1. potassium 3.9-7.6 mg/1. ammonia 0.321-0.93 mg/1. nitrate 1.0-1.99 mg/1. nitrite 0.04-0.09 mg/1, chloride 16.7-27.5 mg/1. sulfate 177-278 mg/1. phosphate 0.035-0.038 mg/1 and sulphide 0.32-0.85 mg/1. The water sample F had 0.068-0.64 mg/1 chromium. 0.35-0.52 mg/1 zinc, 0.38-0.49 mg/1 copper. 0.04-0.074 mg/l nickel and 0.006-0. 141 mg/1 lead.
7 Overall results show that factories are discharging effluents containing high TSS. BOD. COD. sulphides .and nickel into the river. Levels of these parameters exceed the NEQS permissible limits for effluents. Effluents are thus causing sublethal pollution in the river. Some of the parameters like TSS, COD. sulphides. in the downstream river water exceed the NEQS permissible limits during low flow period. Similarly metals in the downstream river also showed an increasing trend when compared with the upstream samples. showing metal stress in the downstream portion of the river.
II.The effects of wastewater pollution on Mahaseer, Tor putitora are summarized below: 1 The total body weight, standard length, total length and some other morphometric parameters decreased.
2 The fish collected from site I and site II. when compared with the control had lower hemoglobin (38% and 12%. respectively). RBC counts (35% and 11 %. respectively), PCV (21 % and 7%. respectively) and MCHC (26% and 20%. respectively) and higher WBC counts (118% and 85%. respectively) and MCV (26% and 13%. respectively).
3 In blood. total protein and cholesterol decreased 47% and 65%. and 51 % and 19%. respectively. in fish collected from the site I and II. respectively. when compared with those of fish collected from control waters. The blood glucose content. GOT and CPK activities of fish collected from site I and II. increased 28% and 29%. 56% and 15%. and 42460% (426 told) and 9318% (932 told). respectively. The GPT activity was increased 124.24% in sample 1 and inhibited 12.12% in sample 2.
4 The hepatic ,total proteins. soluble proteins. total cholesterol. total lipids, glucose content and free amino. acids in fish sample 1 and 2 increased 29% and 16%, 6% and 15%, 75% and 68%, 41% and 65%,47% and 26%, and 333% and 729%, respectively. The DNA content decreased 14% and 20% in sample I and 2, respectively, whereas the RNA decreased 41 % and 32% in sample 1 and 2, respectively. The fish sample from site I and II showed 5% and 17% increased hepatic amylase activity, and 69% increased and 72% decreased GOT activity in sample 1 and sample 2, respectively. The GPT activity decreased 45% in sample 1 and increased 28% in sample 2. The LDH activity was decreased 69% in sample 1, but showed 14% increase in sample 2.
5. The muscle total proteins and total lipids increased 12% and 90%, and 407% and 120%, respectively in sample 1 and 2. The soluble protein contents increased 77% in muscle or fish sample 2. The cholesterol, glucose and free amino acids, however, decreased 17% and 12%, 41 % and 51 %, and 12% and 31 %, respectively, in sample 1 rind 2. The free fatty acids showed 54% lower value in fish sample 1, but 97% higher value in fish sample 2. The DNA content of muscle increased 19% in sample 1 and 2, whereas the RNA contents decreased 24% and 40%, respectively in fish sample 1 and 2. The amylase and LDII activities increased, respectively, 36% and 43%, and 22% and 46% in fish sample I and 2. The GOT and GPT activities of fish muscle showed 19% and 43%, and 46% and 49% lower values in fish sample 1 and sample 2, respectively.
III.The various organs of fish accumulated heavy metals from the environment. 1. In gills zinc was the highest in concentration and chromium was the lowest. The order or metal bioaccumulation was zinc > lead > nickel > copper> chromium. The fish sample from site I and site II had 25% and 14%, respectively, more chromium in the gills than in control fish. Likewise, these fish had respectively, 140% and 150% more nickel. 43% and 46% more lead, 52% and 72% more copper, and 7% and 21 % more zinc in gills of fish sample 1 and 2.
2 The fish liver showed the highest concentration of zinc. while chromium was the lowest. The order of metal bioaccumulation in this organ was zinc > copper> lead> nickel> chromium. The fish sample from site 1 and 2 showed 1300% and 3100% more chromium in liver of fish. Likewise. these fish had 21 % and 26% more nickel. 27% and 57% more lead. 159% and 34% more copper. 36% and 14% more zinc in the livers of fish samples 1 and 2. respcctively.
3 In fish muscle, zinc was the most concentrated metal and chromium was the least. The order of metal bioaccumulation in the muscle was zinc> lead> nickel> copper> chromium. The muscles of fish samples from site 1 and 2 showed 63% and 60% more chromium. 47% and 47% more nickel. 25% and 172% more lead, 19% and 32% more copper, and 57% and 44% more zinc in fish from polluted site 1 and site 2, when compared with those from control waters of Warsak Dam.
4 In fish skin. chromium was the least. while zinc was the most concentrated metal. The order of metal bioaccumulation in the skin was zinc> lead> nickel > copper> chromium. The skin of fish samples from site 1 and 2 showed 13% and 30% more chromium, 67% and 71 % more nickel. 29% and 46%. more lead, 26% and 41 % more copper and 9% and 9% more zinc, when compared with those from control water of Warsak Dam.
IVHeavy metal bioaccumulation 1 The order of chromium bioaccumulation in the different tissues was gills> skin> muscle> liver. The order of nickel bioaccumulation in the different tissues examined was gills> liver> skin> muscle. The order of lead bioaccumulation in the different tissues was gills> skin> liver> muscle. The order of copper bioaccumulation in the different tissues examined was liver> muscle> skin> gills. The order of zinc bioaccull1ulation in the different tissues examined was gills> liver> muscle> skin.
2 Nickel. lead and chromium generally increased in different tissues with the increase in fish age. The concentration of copper also showed an increase in the accumulation tendency with increase in age, except for gills. Zinc showed the same trend in gills, but the trend was different in other tissues. However. overall metal bioaccumulation did not seem to be age and body length dependent.
3 The highest amount of metal bioaccumulation was found in the gills followed by the liver and skin while the lowest was in the muscles. Thus the organ level order or metal bioaccumulation in Mahaseer. Tor putitora was in the sequence gills> liver> skin> muscle.