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Ecological and Microbiological Characteristics of the Jhelum River in Kashmir Himalaya | OMICS International
ISSN: 2155-9597
Journal of Bacteriology & Parasitology

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Ecological and Microbiological Characteristics of the Jhelum River in Kashmir Himalaya

Shugufta Jan1, Imran Khan1,2, Gowhar H Dar1,2*, Azra N Kamili2 and Irfan-ur-Rauf Tak2

1Department of Environmental Science, University of Kashmir, Srinagar 190006, India

2Centre of Research for Development, University of Kashmir, Srinagar 190006, India

*Corresponding Author:
Gowhar H. Dar
Department of Environmental Science
University of Kashmir
Srinagar, 190006 India
Tel: +91-9797124446
E-mail: [email protected]

Received date: April 04, 2016; Accepted date: May 23, 2016; Published date: May 30, 2016

Citation: Jan S, Khan I, Dar GH, Kamili AN, Tak IR (2016) Ecological and Microbiological Characteristics of the Jhelum River in Kashmir Himalaya. J Bacteriol Parasitol 7:277. doi:10.4172/2155-9597.1000277

Copyright: © 2016 Jan S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Abstract

We have assessed the density and diversity of bacterial load of the water body, the Jhelum River from Kashmir Himalaya. Bacteria were isolated and identified and some physical parameters like pH and temperature were measured between June-November 2011, in four sampling sites along the river. The highest bacterial count of was observed at site IV, with a cfu/ml of 2.0 × 103 in the month of July. The lowest viable count was at site III, with a cfu/ml of 0.3 × 103 in the month of November. Among different strains isolated it was found that 73% of strains isolated were Gram negative and about 27% of strains were Gram positive. It was also observed that 5% of strains were Gram positive cocci, 27% strains were Gram negative cocci, 15% were Gram positive Bacilli and 34% were Gram negative Bacilli. It was also found that 32% strains were cocci, 49% Bacilli, 12% strains were Diplococci and 7% strain were Streptococci. Comparative analysis of the different colonies from the four sampling sites indicates that the highest bacterial density was reached in the month of July 2011.

Keywords

Jhelum river; Ecology; Microbiology; Bacteria; Cfu/ml

Introduction

Water is always been mankind’s precious resource and ninety nine percent (99%) of the water on the planet earth is not suitable for human consumption. Most of the water is stored in groundwater. Although water is the most common and important chemical compound on earth, only 2.6% of the global water is freshwater and consequently available as potential drinking water. Availability of sufficient volume of drinking water continues to present major problems, worldwide, owing to the increasing population growth [1]. Also, other complications of highly populated areas, such as increasing amounts of waste, wastewater, and other types of contamination, also endangered access to fresh, safe drinking water [2]. This has led to the development of sophisticated techniques and systems to obtain access to new water reservoirs and to distribute water for irrigation and drinking purposes [3]. Coliforms are a broad class of bacteria that live in the digestive tracts of humans and many animals. Potable or drinking water is defined as having acceptable quality in terms of its physical, chemical, and bacteriologic characteristics so that it can be safely used for drinking and cooking. Water quality standards have been developed to minimize known chemical and microbial risks. The term “safe” drinking water does not mean risk free; it simply means risks are very small, at or below our ability to quantify them, or that water quality limits cannot be lowered further by economical water treatment processes [4]. According to the Bureau of Indian Standard (BIS) IS: 10500 Drinking Water Specification, the official standard for the bacteriological quality of drinking water is: no detectable E. coli, thermo tolerant coliform bacteria, or total coliform bacteria in 100 ml of water. Same standards are given by World Health Organization. Many potential entry points exist, including waste disposal processes (toilet flushing), cleaning (e.g., bathing, hand washing), and indoor/outdoor sanitary activities (e.g., kitchen sink use, storm water collection, irrigation with reclaimed water), and water distribution systems (e.g., fire hydrants, storage tanks, irrigation channels) [5-7]. The intrusion of biological agents into water systems can pose serious public health risks because these agents cannot be easily detected and can remain hidden until a widespread contamination exists. Most cases with known causes have demonstrated considerable delay by authorities to perceive a risk and respond to the situation. For instance, the largest waterborne outbreak in the US resulted in massive illness among 403,000 people in Milwaukee [8]. Ford and MacKenzie noted that water was contaminated for at least 2 weeks before oocysts were identified in stool samples and water contamination was finally suspected [8]. Moreover, the management of aquatic environments requires an understanding of the important linkages between ecosystem properties and the way in which human activities can alter the interplay between the physical, chemical and biological processes that drive ecosystem functioning. Microorganisms play important roles in human daily existence such as fixation of atmospheric nitrogen, decomposition of organic waste and residues, suppression of soil-borne pathogens, degradation of toxicants including pesticides, complexion of heavy metals to limit plant uptake etc. Others, however, are harmful and are called pathogens (disease causing organisms). Some of these pathogens are able to survive both in the soil, air and water ecosystems [9,10]. Itah in 1999 reported that water contaminated by human or animal wastes may harbor pathogens of public health significance depending on the health of the source of the pollution and the degree of treatment [11]. Samra et al. reported the detection of Salmonella sp., Shigella sp., Staphylococcus sp., Streptococcus sp. and Escherichia coli in drinking water sources mainly due to poor water conditions, food and sewage, disposed of into the water bodies [12]. In short, every area of our environment is replete with them. The study of microbes and microbial communities in water environments is broadly known as aquatic micro biology. Microbial communities of aquatic environments include viruses, bacteria, fungi, algae and other microbes. The aquatic systems are mostly dominated by bacteria and fungi and in the natural environments micro-organisms have very specific roles with regard to the recycling of materials and purification of water.

No substantial work has been carried out regarding the current understanding and distribution of bacterial communities in this water body. Therefore the objectives of this study were to identify and isolate the bacteria and to get an idea about the bacterial load (density and diversity) of the water.

Materials and Methods

Study area and sampling sites

The most well-known river of the Kashmir valley is Jhelum River locally known as veth. The Jhelum rises from the spring of Verinag, on the northwestern side of Pir Panjal and flows in a direction parallel to the Indus at an average elevation of 5,500 feet [13]. A total of four sites were selected randomly to access the Bacterial load of the Jhelum River as shown in (Figure 1).

bacteriology-parasitology-Map-showing-study

Figure 1: Map showing study areas.

Site I (Marwal): It was located about 32 km from the main city centre (Lal Chowk) in District Pulwama with geographical coordinate 330 58’ 45.4” N and 740 54’ 16.5” E in rural area, with lots of rural settlements on both the banks. There were also the dense trees of different species especially of Populous and Salix. The average depth of river at the site was about 1.6 meters.

Site II (Aramwari near Zero bridge): This site was located about 1.2 km from the Srinagar city with geographical coordinate 340 4’ 9.2” N and 740 50’ 20.8” E, characterized by congested human population and commercial activities along both its banks. The average depth of river at the site was about 2 meters. All along its course from Marwal to Srinagar the river receives significant quantities of domestic wastes from human settlements.

Site III (Qamarwari): This site was located about 10 km from main city centre with geographical coordinate 340 05’35.9” N and 740 46’45.4” E was having both the commercial activities and residential settlements along both banks, which directly release the sewage and other solid wastes directly into the river. The average depth of river at the site was about 1.2 meters.

Site IV (Tangpora): It was located about 26 km from main city centre with geographical coordinates 340 7’ 47.1” N and 740 43’ 11” E in rural area with human settlements along both the banks. The site was also characterized by the dense vegetation of Populous and Salix and other plants. All along the banks the land was used for the agricultural purposes.

Laboratory analysis

Surface water samples were collected monthly in sterilized bottles from June to November, 2011. The pH and temperature of the samples were recorded on the spot with the help of thermometer and Digital pH meter. The water samples were mixed with sterile distilled water and a series of dilutions were performed. From each dilution, 0.1 ml inoculum was poured onto nutrient agar medium and incubated at 28 ± 2°C for 1 week to assess the growth of colonies [14-21]. Purified isolates were used as stocks for further morphological. Bacterial films were prepared from each purified isolate and stained with Gram’s stain, then examined under the bright field microscope with the oil immersion lens [22]. The number of colonies counted was expressed as cfu/ml and were calculated by using the formula:

cfu/ml=n × d

cfu/ml=n × d

Results

Different types of colonies were obtained during the study period. Some colonies were circular, some irregular and a few colonies were Rhizoid and Filamentous (Table 1). Cocci, bacilli, etc. are different morphological forms of bacteria. A total of 41 strains of bacteria were isolated from the four sites during the study. The different colonies obtained during the study were tested for gram’s stain and subsequently were examined under microscope to determine the Grams nature. The strains isolated with varying colony morphology and their Gram’s tests shown in Table 1 were given codes ranking from B1 to B41. Most of the colonies were circular, Entire, and flat in appearance, margin and elevation respectively. In addition to this circular, Entire and convex colonies were also isolated. Further the strains were tested for Gram’s reaction and it was found maximum strains were Gram negative. After microscopic examination 32% strains were Cocci, 49% were bacilli and the rest of the strains were diplococci and streptococci. The total and individual colony count of different isolates of bacteria isolated from different study sites during the study period are shown in Table 2. The total isolate count reveals that the highest isolate count was B4 (n=149) for all sites followed by B1 with 148;B3 with115;B18 with 87;B10 with 83;B14 with 72; B26 with 62;B11 with 52;B6 and B7 with 49 each;B25 with 48; B29 with 45;B27 with 44;B16 with 43;B8 with 41;B31 with 40;B5 with 39;B15 with 37;B12 with 36;B20 with 34; B22 and B33 with 33 each;B30 with 32;B32 with 28;B38 with 27;B13 and B17 with 25 each;B28 with 24;B19 with 23;B39 with 21;B9 and B35 with 19 each;B21 with 15;B24 with 14;B23 and B34 with 13 each;B37 and B41 with 9 each;B36 with 8;B40 with 7. The individual colony count of different bacterial isolates given in Table 2, reveals that isolate B2 was having the highest number of colonies (52) at site I followed by B1, B4,B3 and others. Among the different isolates obtain, maximum cfu/ml (2.1 × 102) was found in the month of July at site IV (Table 3 and Figure 2). During the study the water temperature and pH recorded at the four sites as shown in Table 3 reveals that a maximum water temperature (22°C) was at site IV in July while it was minimum(5.8°C) at site IV in November. Among different strains it was found that 73% of strains isolated were Gram negative and about 27% of strains were Gram positive (Table 4 and Figure 3). It was also observed that 5% of strains were Gram positive Cocci, 27% strains were Gram negative Cocci, 15% were Gram positive Bacilli and 34% were Gram negative Bacilli (Table 5 and Figure 4). It was also found that 32% strains were Cocci, 49% Bacilli, 12% strains were Diplococci and 7% strain were Streptococci (Figure 5).

S. No. Appearance Margin Elevation Color Grams reaction Cell shape Assigned name
1. Circular Entire Flat White -ve C B1
2. Circular Entire Flat Yellow +ve B B2
3. Irregular Undulate Flat Cream -ve B B3
4. Circular Entire Flat Cream -ve B B4
5. Rhizoid Filamentous Flat Yellow -ve B B5
6. Irregular Undulate Raised Cream -ve C B6
7. Irregular Undulate Flat Yellow +ve B B7
8. Filamentous Filamentous Flat Cream +ve B B8
9. Irregular Undulate Raised Yellow -ve C B9
10 Irregular Undulate Flat Yellow -ve C B10
11 Irregular filamentous Flat Yellow -ve C B11
12 Irregular Undulate Flat Cream -ve C B12
13 Irregular Undulate Flat White -ve B B13
14 Circular Entire Raised Cream +ve B B14
15 Rhizoid Filamentous Flat White -ve C B15
16 Circular Undulate Raised Cream +ve C B16
17 Irregular Filamentous Raised Cream +ve DC B17
18 Circular Filamentous Flat Yellow -ve C B18
19 Rhizoid Filamentous Flat Cream -ve B B19
20 Irregular Lobate Flat Cream -ve B B20
21 Irregular Lobate Raised Cream -ve C B21
22 Circular Entire Elevated White -ve B B22
23 Circular Entire Flat Orange -ve C B23
24 Filamentous Filamentous Flat White +ve B B24
25 Circular Undulate Raised Cream -ve DC B25
26 Irregular Lobate Flat White +ve B B26
27 Circular Entire Convex White -ve DC B27
28 Irregular Filamentous Flat White +ve SC B28
29 Circular Entire Convex Cream -ve B B29
30 Filamentous Undulate Flat Yellow +ve C B30
31 Circular Entire Elevated White -ve DC B31
32 Circular Entire Raised Cream -ve SC B32
33 Filamentous Filamentous Convex Cream -ve DC B33
34 Irregular Undulate Raised Cream -ve B B34
35 Irregular Lobate Flat Cream -ve B B35
36 Circular Entire Convex Yellow -ve C B36
37 Filamentous Elevate Flat Cream +ve DC B37
38 Circular Lobate Flat Yellow -ve B B38
39 Circular Filamentous Flat White -ve B B39
40 Rhizoid Lobate Flat Cream -ve B B40
41 Circular Curved Flat White -ve SC B41

Table 1: Colony morphology and microscopic examination of isolates from four sites.

Isolate number Sites  
Site I SiteII SiteIII SiteIV TOTAL
B1 38 44 30 36 148
B2 52 24 42 28 146
B3 28 23 38 26 115
B4 32 44 40 33 149
B5 3 11 15 10 39
B6 20 05 06 18 49
B7 14 09 07 19 49
B8 16 09 13 03 41
B9 4 09 04 02 19
B10 16 30 13 24 83
B11 11 10 05 26 52
B12 11 23 02 0 36
B13 10 13 02 0 25
B14 08 03 39 22 72
B15 2 03 24 08 37
B16 10 12 15 06 43
B17 09 04 02 10 25
B18 15 11 18 43 87
B19 02 03 12 06 23
B20 07 08 10 09 34
B21 08 0 07 0 15
B22 08 08 09 08 33
B23 03 03 03 04 13
B24 0 03 08 03 14
B25 11 09 03 25 48
B26 04 11 27 20 62
B27 03 0 16 25 44
B28 01 05 06 12 24
B29 06 07 17 15 45
B30 05 06 09 12 32
B31 07 14 08 11 40
B32 07 05 11 05 28
B33 01 02 16 14 33
B34 05 0 03 05 13
B35 05 03 01 10 19
B36 01 04 0 03 08
B37 03 01 01 04 09
B38 03 12 03 09 27
B39 14 02 02 03 21
B40 03 02 0 02 07
B41 01 06 02 0 09

Table 2: Total isolate count of four different sites.

Site June July Oct. Nov.
CC cfu/ml pH T (°C) CC cfu/ml pH T CC cfu/ml pH T (°C) CC cfu/ml pH T (°C)
Site I 132 1.3×103 7.6 17 149 1.5×103 7.6 20.16 54 0.5×103 7.1 11 63 0.6×103 7.0 6.0
Site II 140 1.4×103 7.8 19.1 153 1.5×103 7.7 19 61 0.6×103 7.1 11.9 48 0.5×103 7.1 6.6
Site III 184 1.8×103 7.7 21.7 194 1.9×103 7.7 19 86 0.9×103 7.0 12.3 25 0.3×103 7.1 6.2
Site IV 141 1.4×103 7.7 21 198 2.0×103 7.8 22 104 1.4×103 7.2 11.2 57 0.6×103 7.1 5.8

Table 3: Colony count, cfu/ml at the four sites.

S.NO. Isolate type Gram’s reaction Percentage Cell shape
1 B2 +ve 11(27%) B
2 B7 +ve B
3 B8 +ve B
4 B14 +ve B
5 B16 +ve C
6 B17 +ve DC
7 B24 +ve B
8 B26 +ve B
9 B28 +ve SC
10 B30 +ve C
11 B37 +ve DC
12 B1 -ve 30(73%) C
13 B3 -ve B
14 B4 -ve B
15 B5 -ve B
16 B6 -ve C
17 B9 -ve C
18 B10 -ve C
19 B11 -ve C
20 B12 -ve C
21 B13 -ve B
22 B15 -ve C
23 B18 -ve C
24 B19 -ve B
25 B20 -ve B
26 B21 -ve B
27 B22 -ve C
28 B23 -ve B
29 B25 -ve C
30 B27 -ve DC
31 B29 -ve B
32 B31 -ve DC
33 B32 -ve SC
34 B33 -ve DC
35 B34 -ve B
36 B35 -ve B
37 B36 -ve C
38 B38 -ve B
39 B39 -ve B
40 B40 -ve B
41 B41 -ve SC
  Total 41(100%)

Table 4: Percentage of gram +ve and –ve isolates.

S. No Isolate type  Gram’s reaction Percentage   Cell shape
1 B2 +ve 6 (15%) 20(49%) B
2 B7 +ve B
3 B8 +ve B
4 B14 +ve B
5 B24 +ve B
6 B26 +ve B
7 B3 -ve 14(34%) B
8 B4 -ve B
9 B5 -ve B
10 B13 -ve B
11 B19 -ve B
12 B20 -ve B
13 B21 -ve B
14 B23 -ve B
15 B29 -ve B
16 B34 -ve B
17 B35 -ve B
18 B38 -ve B
19 B39 -ve B
20 B40 -ve B
21 B16 +ve 2(5%) 30 (32%) C
22 B30 +ve C
23 B1 -ve 11(27%) C
24 B6 -ve C
25 B9 -ve C
26 B10 -ve C
27 B11 -ve C
28 B12 -ve C
29 B15 -ve C
30 B18 -ve C
31 B22 -ve C
32 B25 -ve C
33 B36 -ve C
34 B17 +ve 2(5%) 8 (19%) DC
35 B37 +ve DC
36 B27 -ve 3(7%) DC
37 B31 -ve DC
38 B34 -ve DC
39 B28 +ve 1(2%) SC
40 B32 -ve 2(5%)   SC
41 B41 -ve SC
Total 41(100%)

Table 5: Percentage of different bacterial strains.

bacteriology-parasitology-Graphical-representation

Figure 2: Graphical representation of colony counts (cfu/ml) at the four different sampling sites.

bacteriology-parasitology-bacteria-during

Figure 3: Contribution of G+ve and G-ve bacteria during the study.

bacteriology-parasitology-Bacilli-during

Figure 4: Contribution of G+ve, and G-ve Cocci and Bacilli during this study.

bacteriology-parasitology-Contribution-of-different

Figure 5: Contribution of different strains of Bacteria during this study.

Discussion

The total monthly bacterial population as given in Table 3 and Figure 2 showing that the number of bacterial count decreased from July to November 2011. This decrease in the bacterial count may be attributed to the difference in various biotic and abiotic factors that have been found to influence the density and diversity of bacterial communities [17,18]. Another reason for the decrease of bacterial population may be attributed to decreasing pH because the pH varied between 7.2 to 7.8. The present study is confirmed by the findings of Lauber et al. who indicated significance of pH and temperature for the growth of bacterial colonies [23]. Similarly, Dar et al. in 2012 observed that bacterial count declines in October and November with decrease in pH and temperature in soils [18]. The bacterial count was highest at site IV (2.0×103) cfu/ml in the month of July compared to the site III (0.3 × 103) cfu/ml in the month of November, which may be attributed to temperature. Similar results were found by Murphy and Dar et al. 2011, who reported that bacteria grow fast at higher temperature and the growth rate slows down dramatically at low temperature [17,24]. It was found that maximum strains were Gram negative after microscopic examination (32% were Cocci, 49% were bacilli). The results of our study are in consonance with some recent studies on the bacteriological analysis of water and soil in the Kashmir valley [19,21,25]. Also the anthropogenic pressure along the catchments has adversely affected the quality of lotic water systems of Kashmir [13]. The Presence of Gram negative Cocci are of much concern because of their pathogenicity resulting in diseases in humans and the abundance of the gram negative bacteria observed at different sites may be attributed to the increased addition of the sewage and other animal excreta into the river [17]. The aquatic environment is considered a hot-spot for horizontal gene transfer, and lake sediments offer the opportunity for reconstructing the pollution history and evaluating the impacts. The contamination of sediments by untreated or partially treated effluent water can affect the quality of ecosystem [26]. As the gram negative bacteria have a reservoir in the intestines of man and other warm blooded animals, are excreted in feces and are known to survive in the environment but do not reproduce [27]. Thus, the contamination of Water bodies by untreated or partially treated effluent water can affect the quality of ecosystem.

Conclusions

Among the Bacterial strains isolated and identified most of them were Gram- negative bacilli followed by Gram-negative cocci. These are of much concern because of their pathogenicity resulting in diseases in humans. Since the present study is of preliminary nature more investigation in this direction should be undertaken.

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