International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014 122

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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
4
Department of Microbiology, Kohat University of Science and Technology Kohat, Khyber Pakhtoon Khwa, Pakistan.
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

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The present study was formatted to determine the irrigation quality of district Kohat Khyber Pakhtunkhwa of Pakistan. In Ko-
hat 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 irriga-
tion water and Salmonella, Shigella, V.cholerae, Pseudomonas in dams, springs and well water. The Physico-chemical analysis re-
sults 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), Arse-
nic(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, Pseudomo-
nas, 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. Artifi-
cial application of water to the land or soil is irrigation. Water
is used for growing of agricultural crops, landscapes mainte-
nance, and due to water dry land is fit for cultivation of vege-
tables and many other crops especially in those areas in which
rainfall is low[1].
Economy of Pakistan is mostly depending upon irrigation be-
cause climate of Pakistan in some areas is arid and in some
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 charac-
teristics are mainly based on geology and climate of a region.
Geology and climate affect the quality of water directly or in-
directly 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 undevel-
oped countries for irrigation purpose [4]. Global problem is of

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International Journal of Advancements in Research & Technology, Volume 3, Issue 3, March-2014
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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 pur-
pose 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) con-
taminated 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 irriga-
tion 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 con-
tamination and to analyze the Physico-chemical quality of ir-
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rigation water from different sources of Kohat.
2 MATERIALS AND METHOD:
2.1Site Description
Kohat is a medium sized city in Khyber-Pakhtunkhwa Prov-
ince of Pakistan. The Sources for irrigation purposes are main-
ly Tube wells, Wells, springs, Dams and Pressure pumps.
2.2 Water Sample Collection
Sampling was performed from July (2013) to Septem-
ber (2013) and the sampling was done randomly from differ-
ent 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 ac-
cording 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 transpor-
tation to avoid the further contamination.
2.3 Bacteriological Anaylsis
The experiments for bacteriological analysis of irriga-
tion water were carried out at the laboratories of Pakistan
Council Scientific and Industrial Research (PCSIR) complex
Peshawar. Analysis of E. coli, Pseudomonas, Vibrio cholera, Sal-
monella 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
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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 hard-
ness, magnesium hardness, chloride, sodium, potassium, total
suspended particals and heavy metals according to the stand-
ard 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 Spectropho-
tometer for the following metals Zinc (Zn). Copper (Cu) Arse-
nic (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 irriga-
tion water sources are contaminated with E. coli except
tube wells water in which 15 samples shows negative re-
sults 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 salmonel-
la except pressure pumps water.
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 de-
tected 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 ex-
cept pressure pump and tube wells water are less contam-
inated.
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 av-
erage MPN value of Total Coliform bacteria is 100/ml of dif-
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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 aver-
age range of pH is 7.8 and the springs have lowest aver-
age range is 7.
3.5.2Magnesium in Irrigation water
Magnesium in Irrigation Water was deter-
mined 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
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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.
3.5.4Total Hardness in Irrigation water
Total Hardness in Irrigation Water was de-
termined by EDTA method. Total hardness ranged from 100 to
1300 mg/l with average TH is 490 mg/l among different irri-
gation 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.
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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).
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 deter-
mined by EDTA methods. Chlorides ranged from 220 to 590
mg/l with average Chlorides is 338 mg/l among different ir-
rigation 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 de-
termined 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. Fig-
ure 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)
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4 DISCUSSION
Total 40 samples were collected from different irrigation water
sources of Kohat. Samples were collected randomly from dif-
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ferent 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.
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 coli-
form bacteria are widely distributed in nature do not neces-
sarily 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, To-
tal suspended solids, Electrical Conductivity (E.C) ,Calcium,
Magnesium, Copper, Zinc, Arsenic, Iron, Chlorides, Potassi-
um, and Sodium have important role in the quality of irriga-
tion 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].
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The hardness of water is commonly understood property of
water which prevent the lather formation with soap and hard-
ness is due to the presence of Iron, strontium, manganese, car-
bonates, bicarbonates, sulphates, nitrates. In this study area,
the total hardness in water from all the ground water re-
sources 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 192-
350mg/L, excessive concentration causes stomach disorder
and difficulty in washing[12].
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 wa-
ter 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, Po-
tassium, 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 (200-
550) mg/L in dams, wells, tube wells, springs and pressure
pumps respectively. FAO standard are (0.2, 2, 5, 0.2, 300-700,
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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 respective-
ly[3].
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 contami-
nated 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, Pseudomo-
nas 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
129
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: 49-
60(2011).
6. Munir. and Ahmed, M, ‘Ground water quality of Ka-
rachi aquifer for irrigation’, Pakistan of Agric. Res. 24
(1): 141-152.(2010)
7. 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 Sci-
ence 3(4), 489-493. (2004).
8. APHA Standard methods for the examination of wa-
ter 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 fe-
cal 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, In-
dia. pp. 98-99(1968)

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water. Land disposal of solid and liquid materials in bacterio-
logical 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 suita-
ble for irrigation according to the FAO standards. Different
physiochemical variations were found in irrigation water
Sources because these water sources have different site of loca-
tion and soil structure and their sounding area composition
and location. Heavy metals were found in normal range in all
sources of irrigation water.
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. 37-
41, Dec 2013
12. M.khan, M.A. Khawaja and M.Riaz. Physico-
Chemical 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 univer-
sity of Science and Technology, Kohat, Pakistan to provide us
Research Facility.

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