International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 122 BACTERIOLOGICAL AND PHYSICO-CHEMICAL ANALYSIS OF IRRIGATION WATER SOURCES OF KOHAT, PAKISTAN Saadullah1, Hidayatullah1, Muhammad Daud*1, Basreen Akhtar2, Imran khan3, Ghazi Rehman4, Maqsood Qaiser4, Arshad Qayyum4, Waqar Younas4, Allah Nawaz4, Sajad Ahmed4, Zenat Fatima Khattak5, Mirina Sakhi6, Muhammad Mushtaq7 1 Department of Microbiology, Hazara University, Manshera,Khyber Pakhtoon Khwa, Pakistan. 2 Department of Botany, Kohat University of Science and Technology, Kohat, Khyber Pakhtoon Khwa, Pakistan 3 Department of Microbiology, Quaid-e-Azam University, Islamabad, Pakistan Department of Microbiology, Kohat University of Science and Technology Kohat, Khyber Pakhtoon Khwa, Pakistan. 4 5 Department of Pharmacy, Kohat University of Science and Technology, Kohat, Khyber Pakhtoon Khwa, Pakistan. 6 Department of Pharmacy, University of Swabi, Anbar Swabi, Khyber Pakhtoon Khwa, Pakistan. 7 Faculty of Animal Husbandry and veterinary sciences,Agriculture University Khyber Pakhtoon Khwa, Pakistan. * Corresponding Author Muhammad Daud PH# +923349556663 E-mail: Khattaks2010@gmail.com ABSTRACT IJOART The present study was formatted to determine the irrigation quality of district Kohat Khyber Pakhtunkhwa of Pakistan. In Kohat dams, wells, tube wells, pressure pumps and springs water are used for irrigation purpose. All five types (dams, wells, tube wells, pressure pumps and springs) of irrigation water samples were exposed to bacteriological analysis which included, Total Coliform bacteria, Salmonella, Shigella, V.cholerae, Pseudomonas and E.coli. The. The Physico-chemical analysis includes (pH, Electric conductivity, Total hardness, Calcium hardness, Magnesium hardness, Total suspended solids, Iron, Arsenic, and Copper & Zinc). The bacteriological analysis result showed the presence of total Coliform bacteria (TCB) and E.coli in all sources of irrigation water and Salmonella, Shigella, V.cholerae, Pseudomonas in dams, springs and well water. The Physico-chemical analysis results showed that the ranges of pH(7, 6, 7.8, 7.5, 7.3), Electrical conductivity (420, 900, 400, 450, 850uScm1), Sodium (200, 200, 350, 400, 550 mg/L), Potassium (7, 7, 3, 6, 9 mg/L), Total hardness (250, 1200, 200, 400, 300 mg/L), Calcium hardness (200, 1300, 400, 100, 250 mg/L), Magnesium hardness (150, 320, 300, 350, 150 mg/L), Total suspended solids (7, 7, 32 ,6 ,8 mg/L), Arsenic(0.04, 0.06, 0.22, 0.05, 0.01 mg/L), Chlorides (290, 280, 590, 320, 230 mg/L), Zinc(0.08, 0.13, 0.04, 0.04, 0.17 mg/L), Copper(0.05, 0.03, 0.03, 0.02, 0.02.mg/L) and iron (1.8, 2,1.6, 2.7, 0.8.mg/L) respectively in the tube wells, springs, dams, Pressure pumps and well water .Bacteriological results showed presence of Total Coliform bacteria (TCB), Salmonella, Shigella, V.cholerae, Pseudomonas, E.coli. in dams, springs and well water sources that not fit for irrigation .The Physico-chemical analysis showed that all the parameters were found in range of FAO standard in different irrigation water sources except Total hardness were exceed in spring water. Keywords : Bacteriological, Salmonella, Shigella, Physico-chemical 1 INTRODUCTION W ater is very Important resource on Earth. On earth it is a valuable gift of nature to mankind and to other living organisms. The accessibility of water has always been of great importance for life, human health, most ecological systems, food production and economic development. Artificial application of water to the land or soil is irrigation. Water is used for growing of agricultural crops, landscapes maintenance, and due to water dry land is fit for cultivation of vegetables and many other crops especially in those areas in which rainfall is low[1]. Economy of Pakistan is mostly depending upon irrigation because climate of Pakistan in some areas is arid and in some Copyright © 2014 SciResPub. areas it is semi-arid. In world irrigation system is largest it consist of dams, canals, rivers, barrages, water reservoir which occupied large area on earth. The water which is available doesn’t meet the requirements of irrigation [2].These characteristics are mainly based on geology and climate of a region. Geology and climate affect the quality of water directly or indirectly by chemical and physical structure of soil including rocks weathering, lime dissolution, gypsum and minerals which are slowly dissolve in soil. On local level there is difference in water quality depending on the source [3] . 80% of polluted water is used in undeveloped countries for irrigation purpose [4]. Global problem is of IJOART International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 polluting surface water (ponds, lakes and rivers) and ground water which in result contamination of the fresh fruits and vegetables. Fecal pathogens such as salmonella, Shigella and E.coli are involved in the fecal contamination of water linked to excessive nutrient levels in irrigation waters. In addition to biological contamination heavy metal is present in drainage water which adds pollution to food chain in addition to ground water contamination used in irrigation of lands [5]. Irrigation water is contaminated by animal feaces which show the presence of E. coli [2, 5, 1].The waste material is enrich with organic matter and plants nutrients, it also contain little amount of soluble salts and heavy metal like copper, zinc ,iron etc. When this contaminated water is used for irrigation purpose for longer period of time so with the passage of time these heavy metals accumulates in the soil and later on it causes toxicity to plants [6].Irrigation with As (arsenic) contaminated ground water to rice & other crops may increase its concentration & eventually As (arsenic) may enter into food chain through crop uptake [7].To know the quality of irrigation water we have to check its hardness, chloride, sodium, copper and calcium. When their concentration is higher so it cause corrosion in distribution system. The aim of the study is to assess the irrigation water quality based on bacterial contamination and to analyze the Physico-chemical quality of irrigation water from different sources of Kohat. 123 the dust and dirt were removed. For sterilization, all glassware and medium were autoclaved at 121°C temperature for 15 minutes at pressure of 15-lb/ inch2 (psi). After autoclaving, all the glassware were dried in oven at 60°C temperature before use. 2.4 Physico-chemical Anaylsis Physico-chemical parameters include determination of PH, electrical conductivity, total hardness, calcium hardness, magnesium hardness, chloride, sodium, potassium, total suspended particals and heavy metals according to the standard of (APHA, 2005)[8]. 2.5 Pre-treatment of Samples All the samples were first filtered before subjecting to the instrument. No pre-concentration step was performed. The samples were analyzed by Atomic Absorption Spectrophotometer for the following metals Zinc (Zn). Copper (Cu) Arsenic (As) iron (Fe) the concentrations were in mg/l. 3 RESULTS 3.1E.coli and Salmonella E. coli Culture was detected on EMB agar (Eosin Methylene Blue). E. coli on EMB Agar was showing green metallic sheen, smooth, regular and transparent colonies. Figure 4.2 shows that all the samples of irrigation water sources are contaminated with E. coli except tube wells water in which 15 samples shows negative results and less contaminated. Salmonella was detected on SS agar. Salmonella on SS agar was showing black centre, irregular and smooth colonies. Figure 4.4 shows that all the sources of irrigation water were positive for salmonella except pressure pumps water. IJOART 2 MATERIALS AND METHOD: 2.1Site Description Kohat is a medium sized city in Khyber-Pakhtunkhwa Province of Pakistan. The Sources for irrigation purposes are mainly Tube wells, Wells, springs, Dams and Pressure pumps. 2.2 Water Sample Collection Sampling was performed from July (2013) to September (2013) and the sampling was done randomly from different locations of Kohat. Forty (40) samples were collected from home wells (HW), tube wells (TW), springs (SP), pressure pumps and dams. The sample collection was performed according to standard method given by (APHA 2005). Samples were collected aseptically in 500 ml sterile autoclavable plastic (polypropylene) bottles and were transferred in an ice box to the laboratory. The samples were air tight during the transportation to avoid the further contamination. 2.3 Bacteriological Anaylsis The experiments for bacteriological analysis of irrigation water were carried out at the laboratories of Pakistan Council Scientific and Industrial Research (PCSIR) complex Peshawar. Analysis of E. coli, Pseudomonas, Vibrio cholera, Salmonella and Shigella were performed in the irrigation water. 2.4 Sterilzation of glass ware and medium All the glass wares like Petri-plates, flasks, pipettes, beakers and test tubes used in experiments were properly washed with available commercial detergent. After scrubbing with a brush, the glassware were rinsed repeatedly with water and then rinsed two or three times in distilled water until all Copyright © 2014 SciResPub. 3.2Shigella and Vibrio Cholerae Shigella was detected on SS Agar. Shigella on SS agar showed light, colorless, transparent and smooth colonies. Figure 4.6 shows that all the irrigation water samples sources were positive for Shigella except tube well and pressure pumps water. Vibrio cholerae was detected on TCBS Agar (Thiosulfate-citrate-bile salts-sucrose agar). Vibrio cholerae on TCBS Agar showed yellow color, irregular, rough and transparent. Figure 4.8 shows that all the irrigation water sources are positive for V.cholerae except pressure pump and tube wells water are less contaminated. 3.3Pseudomonas Pseudomonades were detected on Pseudo Agar. Pseudomonas on Pseudo agar showed light green, transparent and smooth colonies. Figure 4.10 that all the sources of irrigation water are positive for Pseudomonas except pressure pumps and tube wells water. 3.4Total Coliform Bacteria Detection of Total Coliform Bacteria was performed on lactose leuryle broth. Results (Figure 4.11.) shows that average MPN value of Total Coliform bacteria is 100/ml of difIJOART International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 ferent source of Irrigation water of Kohat. The Highest value is 619.25 of Dams Water and the lowest value is 7.33 in pressure pump water. 3.5Physico-chemical Anaylsis 3.5.1pH PH of Irrigation water was determined by pH meter. The pH ranged from 7 to 7.8 with average pH 7.4 among different irrigation water sources was analyzed. Figure 4.12 shows that the dams water has highest average range of pH is 7.8 and the springs have lowest average range is 7. 3.5.2Magnesium in Irrigation water Magnesium in Irrigation Water was determined by EDTA method. The magnesium ranged from 180 to 370 mg/l with average magnesium is 274 mg/l among different irrigation water sources were analyzed. Figure 4.13 shows the results that pressure pumps, spring and Dams water have high average range of magnesium which is 370 mg/l and the tube wells and wells water have low average range which near about 180 mg/l. 3.5.3Sodium In Irrigation water Sodium in Irrigation Water was detected by flame photometer. The sodium ranged from 190 to 540 mg/l with average Sodium is 341 mg/l among different irrigation water sources were analyzed. Figure 4.14 shows that wells water have high ranges of sodium which is 540 mg/l and springs water have lowest average ranges 190 mg/l. 124 3.5.7Arsenic in Irrigation water Arsenic in Irrigation Water was detected by Atomic absorption spectrophotometer. Arsenic ranged from 0.02 to 0.23 mg/l with average Arsenic is 0.08 mg/l among different irrigation water sources were analyzed. Figure 4.18 shows that high ranges are (0.23 mg/l) in dams water while the lowest ranges (0.02 mg/l) in wells water. 3.5.8Zinc in Irrigation water Zinc in Irrigation Water was detected by Atomic absorption spectrophotometer. Zinc ranged from0.03 to 0.17 mg/l with average zinc is 0.09 mg/l among different irrigation water sources were analyzed. Figure 4.19 shows that the springs and wells water has high ranges 0.17- 0.13 mg/l and dams and pressure pumps water 0.06-0.03 mg/l. 3.5.9Total suspended Solids in Irrigation water Total Suspended Solids in Irrigation Water was determined by filtration methods.TSS ranged from 5 to 32 mg/l with average TSS is 12.2 mg/l among different irrigation water sources were analyzed. Figure 4.20 shows that dams water have high ranges of TSS (32 mg/l) and other sources have low range (5-9 mg/l). IJOART 3.5.4Total Hardness in Irrigation water Total Hardness in Irrigation Water was determined by EDTA method. Total hardness ranged from 100 to 1300 mg/l with average TH is 490 mg/l among different irrigation water sources were analyzed. Figure 4.15 shows the results that the springs water have high ranges of TH 1300 mg/l and dams water have lowest ranges of TH 100 mg/l. 3.5.5Potassium in Irrigation water Potassium in Irrigation was determined by flame photo meter. The potassium ranged from 3 to 9 mg/l with average Potassium is 6.6 mg/l among different irrigation water sources were analyzed. Figure 4.16 shows that the wells water springs and pressure pumps water have high average ranges are 6mg/l to 9mg/l and the lowest are in Dams water are 3mg/l. 3.5.6Copper in Irrigation water Copper in Irrigation Water was determined by atomic absorption spectrophotometer. The copper ranged from 0.03 to 0.06 mg/l with average Copper is 0.03 among different irrigation water sources were analyzed. Figure 4.17 shows the results that the tube wells water have high ranges of copper which is 0.06 mg/l and wells water have lowest ranges which is 0.03 mg/l. Copyright © 2014 SciResPub. 3.5.10Iron in Irrigation water Water Iron in Irrigation Water was detected by Atomic absorption spectrophotometer. Irons ranged from 0.6 to 2.6 mg/l with average iron are 1.7 mg/l among different irrigation water sources were analyzed. Figure 4.21 shows that pressure pumps water has high ranges (2.6 mg/l) and wells water has lowest ranges (0.6 mg/l). 3.5.11Chlorides in Irrigation water Chlorides in Irrigation Water were determined by EDTA methods. Chlorides ranged from 220 to 590 mg/l with average Chlorides is 338 mg/l among different irrigation water sources were analyzed. Figures 4.22 shows that dams water have high ranges (590 mg/l) and wells water have low ranges (220 mg/l). 3.5.12Calcium hardness in irrigation water Calcium hardness in Irrigation Water was determined by EDTA methods. Calcium hardness ranged from 100 to 1400 mg/l with average Calcium hardness is 470 mg/l among different irrigation water sources were analyzed. Figure 4.23 shows that spring water have high ranges (1400 mg/l) and pressure pumps water have low ranges (100mg/l). 3.5.13Electric conductivity of irrigation water Electric Conductivity of Irrigation Water was determined by EC meter. Electric Conductivity ranged from 400 to 820 μScm-1 with average EC 584 μScm-1 among different irrigation water sources were analyzed. Figure 4.24 shows that spring and wells water have ranges between (850-820 μScm-1) and dams water have low range (400 μScm-1) IJOART International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 125 IJOART Copyright © 2014 SciResPub. IJOART International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 126 IJOART Copyright © 2014 SciResPub. IJOART International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 127 IJOART Copyright © 2014 SciResPub. IJOART International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 4 DISCUSSION Total 40 samples were collected from different irrigation water sources of Kohat. Samples were collected randomly from different location and from different sources such as dams, wells, tube wells, springs and pressure pumps that it may represent the overall irrigation water sources of District Kohat. 128 The hardness of water is commonly understood property of water which prevent the lather formation with soap and hardness is due to the presence of Iron, strontium, manganese, carbonates, bicarbonates, sulphates, nitrates. In this study area, the total hardness in water from all the ground water resources ranges between 200 and 1300 mg/L and the FAO range are 300-400 mg/L. The total hardness in water from all the ground water resources ranges between 228 and 644mg/L. (Khan etal 1999) concluded the result which ranges from 192350mg/L, excessive concentration causes stomach disorder and difficulty in washing[12]. IJOART In worldwide different disease occur because of pathogens present in irrigation water. In irrigation water pathogen which are recorded they are E. coli, Salmonella, Shigella, Pseudomonas, Vibrio cholerae and Total coliform bacteria. E. coli were detected in all the water samples collected from dams, tube wells ,wells, springs and pressure pumps Total Coliforms were also recorded in this study. Total coliforms were found in all the samples and has high amount in dams water followed by wells. (Griffith et. al., 2003) concluded that Total coliforms were apparently present but it is important to note that coliform bacteria are widely distributed in nature do not necessarily specify fecal pollution [9]. Shigella and Salmonella were founded in three sources dams, wells and springs except pressure pumps and Tube wells while as Vibrio cholerae found in three sources samples except tube wells and pressure pumps. Physicochemical parameters such as pH, Total Hardness, Total suspended solids, Electrical Conductivity (E.C) ,Calcium, Magnesium, Copper, Zinc, Arsenic, Iron, Chlorides, Potassium, and Sodium have important role in the quality of irrigation water. PH of water samples were in the range of 7-7.8.The highest value of water pH that is of Dams water with mean ranges of 7.8. And the FAO ranges are 4.5-9.0. (Ayers and Westcott, 1985) recommended the pH of water samples varied from 6.71 to 7.84 indicating slightly acidic to slightly alkaline in nature and was within the safe limit for irrigation. The recommended pH limit of irrigation water is 6.0-8.5 [3]. Copyright © 2014 SciResPub. The ability of an aqueous solution to carry an electric current is called Electrical Conductivity. In this present study dams water has low range with 400μScm-1 and highest range of spring water with value of 850μScm-1 and the FAO range are 800-1000 μScm-1. (Richards 1968) concluded that EC values of water samples were ranged from 196 to 483 μScm-1[10]. Total suspended solids analysis have ranges of 6mg/L to 34 mg/L. The highest value is 34mg/L of dam’s water while pressure pumps have low value of 6 mg/L. Total Suspended solids ranges from 584 mg/L and lower 380 mg/L. (H.Iqbal et al; 2013) reported total suspended solids which ranges from 10-20mg/L[11]. Calcium and magnesium was found in all the samples having the ranges 200, 150, 400, 1400mg/L, and100, 190,300,150,320 and 350mg/L in wells, tube wells, dams, springs and pressure pumps respectively. The FAO range are 100-1500 mg/L in spring water more Ca and Mg is present as compare to surface water. (H.Iqbal et al: 2013) reported Ca and Mg content of water samples they ranges from 121.1-229.1 to 45.2-111.6 and mg/L respectively[11]. In this study Copper, Zinc, Arsenic, Iron, and Chlorides, Potassium, and Sodium were also analyzed respectively in all the sources of irrigation water. The Ranges of Cu are (0.02 -0.06), Zn (0.04 -0.15), Fe (0.8-2.6),Cl (250-590), K (04-09) and Na (200550) mg/L in dams, wells, tube wells, springs and pressure pumps respectively. FAO standard are (0.2, 2, 5, 0.2, 300-700, IJOART International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 ISSN 2278-7763 8-10,500-600) mg/L. (Ayers and Westcott, 1985) reported Fe; Zn, Mn and Cu contents of water samples were from 0.047 to 2.718, 0.001 to 0.090, 0 to 2.098 and 0 to 0.003 mg/L respectively[3]. 5. Kumasi CT, Kwasi O, Ephraim JH. Microbial quality of water in Barekese reservoir and feeder streams in Ghana. Lakes and Reservoirs: Res. Manage., 16: 4960(2011). 6. Munir. and Ahmed, M, ‘Ground water quality of Karachi aquifer for irrigation’, Pakistan of Agric. Res. 24 (1): 141-152.(2010) Islam, M.R, Jahir Uddin, M. and Islam, S. ‘Assessment of Arsenic in the water-soil-plant systems in Genetic Floodplains of Bangladesh’, Asian Journal of Plant Science 3(4), 489-493. (2004). 5 CONCLUSION Forty (40) samples were collected from the different sources of water (dams, wells, tube wells, springs and pressure pumps) of kohat and bacteriological and physicochemical analysis was done. From the present study it is concluded that the tube wells and pressure pumps water has less bacterial contaminated and Dams, wells, and springs are highly contaminated with bacterial pathogens, like E. coli, Total coliform bacteria, Salmonella, Shigella, Pseudomonas and vibrio cholerae. Tube wells water and pressure pumps water are not exposed to external environment, more deeper , passes through different layers of soil. Wells water, dam’s water and springs water are more contaminated and showed positivity of different bacteria like E. coli, Total Coliform bacteria, Salmonella, Shigella, Pseudomonas and vibrio cholerae. Because these are exposed to external environment. Runoff is not control .Easily contaminate the exposed water through fecal, human and animal waste on the land surface problems Infiltration of contaminated surface water. Land disposal of solid and liquid materials in bacteriological point of view the exposed water sources are not fit for irrigation Dams, wells, and springs exceeds the FAO standard. Physicochemically all the sources of irrigation water are suitable for irrigation according to the FAO standards. Different physiochemical variations were found in irrigation water Sources because these water sources have different site of location and soil structure and their sounding area composition and location. Heavy metals were found in normal range in all sources of irrigation water. 129 7. 8. APHA Standard methods for the examination of water and waste water, Washington, D. C. American Public Health Association. 21st Edn(2005), 9. Griffith, J. F, Weisberg, B. S., McGee, D. C.Evaluation of microbial source tracking methods using mixed fecal sources in aqueous test samples. J. Water Health. 1:141-151(2003). 10. Richards, L.A. ‘Diagnosis and Improvement of Saline and Alkali Soils’, Agricultural Hand Book 60(ed), USDA and IBH. Publishing Co. Ltd. New Delhi, India. pp. 98-99(1968) IJOART 11. H.Iqbal, M.Ishfaq, A.Jabbar, M.Naseer Abbas, A.Rehaman, S.Ahmad, M.Zakir, S.Gul, B.Ibtesam Shagufta, M. Ullah, W.Ahmad. Physico-Chemical Analysis of Drinking Water in District Kohat, Khyber Pakhtunkhwa, Pakistan. IJBMSP, Vol. 3, No. 2, pp. 3741, Dec 2013 12. M.khan, M.A. Khawaja and M.Riaz. PhysicoChemical Characteristics of Sub-Surface water of Haripur area-Hazara (N.W.F.P).Jour. chem. Soc. Pak. Vol 21,No 4,1999 6 ACKNOWLEDGMENT We are thankful to Deparment of Microbiology Kohat university of Science and Technology, Kohat, Pakistan to provide us Research Facility. 7 REFERENCES 1. Solomon EB, Yarn S, Mathews KR .Transmission of Escherichia coli 0157:H7 from contaminated Mann and irrigation water to lettuce plant tissue and its subsequent internalization. Appl. Environ. Microbial. pp. 397-400(2002). 2. N. Ahmad and G. R. Chaudhry Irrigated Agriculture of Pakistan Lahore, (1986). 3. Ayers, R.S. and Westcott, D.W, ‘Water Quality for Agriculture’ FAO Irrigation and Drainage Paper. 29Rev (I): 8-96. 1985 4. D. Mara, and S. Cairn cross, Guidelines for safe use of wastewater and excreta in agriculture and aquaculture: measures for public health protection. World Health Organization, Geneva, pp.187, 1989. Copyright © 2014 SciResPub. IJOART