Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 624-631

ISSN: 2319-7706 Volume 4 Number 7 (2015) pp. 624-631 http://www.ijcmas.com

Original Research Article Diversity and Density of Coliform Bacteria in River Tunga at Shivamogga city, Karnataka,

S. Prathibha and V.N.Murulidhar*

1Department of Botany, JSS Degree College for Women (Autonomous), Mysore-570025, Karnataka, India 2Department of Botany, Government First Grade College, Sira-572103, Tumkur District, Karnataka, India *Corresponding author

A B S T R A C T

Water resources are being polluted due to increasing urbanization, modern practices in agriculture and other human activities. The link between human health K e y w o r d s and environmental pollution is of significant dimension. Studies were conducted with an objective of assessing the coliforms density and dynamics in river Coliform Tungawater. River is polluted with domestic and agriculture wastes of Shivamogga bacteria, city. Water samples were collected from four designated sampling stations of the Water, river. Results revealed that, all the samples from the selected stations contained River, coliforms bacteria above the safe limits for drinking water standards. Thus river Pollution water is unsatisfactory as judged by mean MPN counts of indicator bacteria i.e., total coliforms 2.22 x 107/100 ml. Seasonal variations in coliforms density were recorded with maximum density in summer and monsoon seasons and minimum density in winter seasons.

Introduction

Water contamination by various routes and directly to detect the presence of all the agents are liable to cause clinically signified different kinds of water borne pathogens, outbreaks of intestinal infections in both any of which may be present only animals and man. The demonstration of intermittently. Thus, we rely on tests that pathogenic bacteria, would obviously reflect the presence of commensal bacteria constitute the most direct proof of a of intestinal origin such as those of the hazardous impurity; but these pathogens, if coliform group, which are more numerous, present are so scanty that the technical more easily tested and are the most reliable difficulty of their isolation makes the test indicators of fecal pollution (Senior, 1989). impractical for ordinary purposes and they Each person discharges 100 to 400 billion are very sensitive even to a slight change of coliform organisms/day, in addition to other environmental factors and difficult to kinds of bacteria. These coliforms do not isolate. Hence, it is impractical to attempt themselves constitute a hazard since, they 624

Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 624-631 are harmless to man and are in fact useful in be indicators for potential pathogens destroying organic matter in biological responsible for various waterborne diseases waste treatment processes. But they indicate (Hunter et al., 2004) or be pathogens that faecal matter has entered and that the themselves, such as certain strains of E. coli. water is therefore, liable to contamination In the present experiment we tested water with more dangerous organisms (Senior, pools in 96 samples at a stretch of 15 km of 1989). Presence of coliform organisms is Tunga River for abundance and species thus taken to indicate possible presence of richness of coliform and other pathogenic organisms. environmental bacteria. We hypothesized that river with more human or animal Coliform monitoring being rapid, anthropogenic activities or watershed with inexpensive easy to perform, proved agricultural or sewage runoff would have effective (Leclerc et al., 2000). Coliform higher bacterial abundance and species was the term first used in 1880 s to describe richness. rod-shaped bacteria. Coliform group of bacteria is functionally-related groups Material and Methods belong to a single taxonomic family Enterobacteriaceae and comprises many The present study area covered a stretch of genera and species. The term coliform 15 km of the river Tunga near Shivamogga organisms refers to gram-negative, rod- city. Ninety six water samples from Tunga shaped bacteria capable of grow in presence river were collected during April 2010- of bile salts or other surface-active agents March 2012 (24 months) from all the four with similar growth-inhibiting properties designated stations and were analysed for and able to ferment lactose at 35 ºC 37 ºC the levels (MPNs) of generally accepted with the production of acid, gas and faecal pollution indicators i.e., total aldehyde within 24-48 hours (Clesceri et al., coliforms (TC). Seasonal variations were 1998). They are also oxidase-negative and examined among these indicators during non-spore-forming and display b- April 2010 to March 2012 as well as the galactosidase activity (Arora, 2003). species composition for each group of Traditionally coliform bacteria were indicators. The pollution range, maxima, regarded as belonging to the genera minima, standard deviation of the above Escherichia, Citrobacter, Enterobacter and parameter has been given along with the Klebsiella. MacConkey (1909) defined 128 detailed discussion of the parameter coliform types and this number was including ANOVA results. Presumptive increased to 256 by a system developed by coliform count: Single and Double strength Bergey and Deehan (1908). However, as MacConkey s broth medium (Hi-media, defined by modern taxonomic methods, the Ltd.) was prepared by dissolving 4 g and 8 g group is heterogenous consisting of as many of MCB in 100 ml of distilled waters as 32 genera (Leclerc et al., 2001). Some of separately. Measured amounts were these can be found in both faeces and the sterilized in test tubes containing a environment (nutrient-rich waters, soil, Durham s tube for indicating gas decaying plant material) as well as in production. With sterile graduated pipettes drinking water with relatively high or with adjustable micropipettes that can concentration of nutrients and may multiply. hold sterilized microtips, the following Eg. Serratia fonticola, Rahnella aquatilis, amounts of the sampled waters were added. Buttiauxella agrestis. Coliform bacteria can

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a. Five 10 ml quantities each to DS between station and season is also (Double Strength) medium. statistically non-significant as all the b. Five 1 ml quantities each to 10 ml SS obtained values failed to reach the (Single strength) medium. significant level. Even though, we find c. Five 0.1 ml quantities each to 10 ml comparative differences among means, these SS medium. mean differences are statistically insignificant, as there were high individual Tubes that show acid and sufficient gas to variations in the data, which is evident from fill the apex covering the concavity of the respective standard deviation (SD) Durham s tube at 24 hours were considered values (Table 2). Presence of coliform to be presumptive positive, as a result of the organisms in water has been attributed to growth of coliform Bacilli after incubation influxes of allochthonous bacteria from at 37 ºC. Incubation was sustained from the waste discharges and surface water drainage. next session; tubes showing presumptive Total coliform levels were always higher positive at 48 hours of incubation were than faecal coliform levels, which is not accredited to the existing numbers in the surprising since total coliforms can originate concerned station. Sample dilutions were from non-faecal sources also such as plants read separately and the positives in A, B and and soils (Geldreich, 1974). The above C were serially collated in probability tables findings and opinions also were to obtain presumptive coliform Bacilli in corroborated by Goyal et al. (1977). 100 ml of the sampled station. Tubes showing gas production at 37 ºC for 48 In the present study, high coliform load has hours were confirmed by plating in been registered during the entire course of MacConkey agar plates (MCA) and study. The MPN of coliform organisms examined for typical colonies. fluctuated from 17 x 101 to 16 x 108 / 100 ml. The higher values of MPN for coliforms Results and Discussion were recorded at TS3 and TS4, mainly attributed to the domestic sewage and In the present investigation two-way municipal wastes. Though, the values ANOVA was employed taking test stations remain statistically significant, due to high (TS1, TS2, TS3 and TS4) as first independent individual variations in the data, as per variable and seasons (summer, rainy and values in the current study the distribution of winter) as second independent variable total coliforms MPN was found bimodal against all parameters considered as with summer and monsoon maxima. Higher dependent variables. The first variable coliform levels occurring during summer (stations) varied at 4-levels, 2nd variable months suggest survival and high influx of (seasons) varied at 3-levels to make 4x3 the coliforms and also increased nutrient design. This will yield different between the influx from decomposition of summer stations, difference between season s productivity because of high temperature interaction between stations and seasons and watershed drainage may also contribute (Lateef et al., 2005). The maximum, to March-April increase in numbers of both minimum and averages of two years in the the years. In addition, heavy usage of the total Coliforms are represented in table 1. In river was found during summer for bathing, the current study, no significant difference washing and such other anthropogenic was noticed between stations and seasons, in activities, that also would contribute for the total coliforms and the interaction effect higher incidence of coliforms. Water quality

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Int.J.Curr.Microbiol.App.Sci (2015) 4(7): 624-631 of Tunga river showed significant bacterial numbers in river water after rain deterioration in view of global standards. All and during summer. This is supported by most all the samples were contaminated with Raj Kumari Sing et al. (2001) who carried coliform bacteria, resulting mainly due to investigations in at anthropogenic activities, especially Hoshangabad. In Lake Windermere, Taylor discharging of domestic and agricultural (1940) has observed that there is a close wastes directly into the river as some recent relationship between the bacterial content of studies (Sharma et al., 2010; Sadat et al., the water and the rainfall in the drainage 2011) revealed that coliform count has area during the preceding week. positive relation with anthropogenic Subramanyan and Bhaskaran (1948), activities. Present results are in agreement Voelkar et al. (1960) noted a considerable with the findings of Geldrich (1972) who decrease in the number of coliforms in dug observed an increase in the total coliform wells during cold months. count of water bodies due to increased use of animal wastes as manure in the Further, TS2 and TS3 have bathing ghats, agricultural fields. where disturbance due to human activities is seen throughout the year. Highest incidence In contrast, Goyal et al. (1977), Sayler et al. was recorded in TS4 followed by TS3, TS2 (1975) found higher organisms in winter and TS1. Coliform load in river water months than in summer. In the present study indicated that bacteria were always and summer maxima was followed by rainy under all conditions remains in water body season. Feachem (1974), Gerba and (Agarwal, 1993). High counts of bacteria Schaiberger (1975) have also reported an signified the organic pollution in river water increase in the number of indicator of Tunga. Extremely high MPN values of organisms and Faechem (1974) found peaks Total coliforms i.e., 1600x1010 were of coliforms during on immediately after a reported near Opium factory in Ganga water storm and other times due to unexplained (Shukla Suresh et al., 1992). They also factors. Occurrence of coliforms in the quote the observations of other workers that samples is also confirmed by study there is a direct effect of sewage containing conducted by Lateef et al. (2003) reporting faecal matter on the growth and population high coliform counts in the spring water of of coliform bacteria. Agarwal (1993) Kashmir valley. Due to change in registered a range of 19-22000/100 ml of TC agricultural methods, diminishing of in river Betwa. Kataria et al. (1997) studied livestock farms, intensive farming operation river Halali and reported 240 to 2400/100 increased concentration of animal wastes ml. Doctor et al. (1998) recorded MPN results in an increased pollution of rivers between 300 to 1600/100 ml in river Bhadar. and streams (Gelt, 1998). High coliform Raka et al. (1999) registered the faecal incidence is influenced by the entry of pollution in drinking water of surface runoff water from nearby areas that and reported the coliform in range of 21 carried higher levels of suspended matter 180/100 ml. Koshy Mathew and Vasudevan and nutrients on their surface. Similar Nayar (2000) point out that, MPN of Pamba observations were made by Venkateswarlu river water was infinite at four of the seven (1986) in the rivers of Andhra Pradesh. stations considered due to Sabarimala According to Keller (1960) access of soil pilgrimage and free flow of sewage, and organic matter and increase in domestic waste and faecal matters into the temperature is responsible for the increase in river at those stations.

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Table.1 Sectoral range and mean values of total coliforms (TC/100ml)

Sampling Mean Std. Deviation Std. Error Minimum Maximum Stations TS1 4252.5000 11273.1402 2301.1201 170.00 54000.00

TS2 1464.1667 1317.8504 269.0051 220.00 5400.00

TS3 450541.67 588224.0019 120070.72 13000.00 1600000

TS4 88260000 324878798.22 66315607 940000.0 160000000

Total 22179065 164390697.01 16778055 170.00 160000000

Table.2 Sectoral and seasonal variation of total coliforms (TC/100 ml) with ANOVA results

Seasons TS1 TS2 TS3 TS4 Overall Mean SD Mean SD Mean SD Mean SD Mean SD Summer 7.39x103 1.88x104 8.7x102 4.15x102 3.66x104 2.15x104 2.36x108 5.54x108 5.90x107 2.83x108

Rainy 4.85x103 5.88x103 1.74x103 1.76x103 1.06x106 6.50x105 2.64x107 4.53x107 6.87x106 2.44x107

Winter 5.16x102 4.58x102 1.79x103 1.35x103 2.53x105 2.19x105 2.52x106 2.75x106 6.93x105 1.69x106

Overall 4.25x103 1.13x104 1.46x103 1.32x103 4.51x105 5.88x105 8.83x107 3.25x108 2.21x107 1.64x108 F for stations (A) = 1.808, P=0.152 (Non-significant); F for seasons (B) = 1.273, P=0.285 (Non-significant); Interaction F (AxB) = 1.281, P=0.275 (Non- significant). Mean values followed by different letters are significantly different from each other.

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They also indicate an increased coliform It is concluded from the present results that, content at the sides of the Pampa River existence of both non-faecal bacteria that fit (1600/100 ml) and very low values at the the definitions of coliform bacteria and of middle part (12/100 ml). They attribute this lactose-negative coliform bacteria limits the to the good velocity at middle with no flow applicability of this group as an indicator of near the sides due to sand mining. Shafi et faecal pollution. Drinking directly from river al. (2013) observed that, all the samples without filtration should be avoided at all obtained from the Manasbal Lake of times. Many organisms present in river are Kashmir were positive with respect to very sensitive to ecosystem. Coliform coliform occurrence, though the count was bacteria should not be detectable in treated variable ranging between 4 and 460 water supplied and if found, suggest MPN/100 ml highest proportion of indicator inadequate treatment, post treatment coliforms was found in water samples contamination or excessive nutrients. collected at the site surrounded by Coliform test can therefore be used as residential hamlets in comparison to other indicator for both treatment efficiency and three sites. Debashish Mandal and integrity of distribution system. Although Mukhyopadhyay (1991) also report coliform organisms may not always be extremely high MPN ranging from 10x102 directly related to the presence of faecal to 9x106/100 ml in river Ganga. According contamination or pathogens in drinking to them coliform bacteria are more abundant water, coliform test is still useful for in certain sewage channels and bathing monitoring the microbial quality of treated ghats. Musaddiq Mohammed (2000) piped water supplies. If there is any doubt, recorded a maximum MPN of 900 in Morna especially when coliform organisms are River and suggests health education as an found in the absence of thermo tolerant important weapon in creating desire for high coliforms and E. coli, identification to the standard of life. Rajkumar Singh et al. species level or analyses for other indicator (2001) noted MPN index up to 9200/100 ml organisms may be undertaken to investigate in Narmada River signifying the organic the nature of contamination. Sanitary pollution in the water body due to the human inspections will also be needed. activities. Asthana and Sing (1993) observed extremely high MPN values of 21x103 to References 40x103 in Gomati River with faecal coliform accounting for nearly 60 to 80% of the total Agarwal, A. 1993. Studies on physico coliform. Praveen Jain and Sanjay Telang chemical and biological characteristics (1996) have shown an MPN of 118 in of river Betwa from Nayapura to Parbathy River at Sehore (M.P) and Vidisha. Ph.D. Thesis, Barkatulla conclude that river water has minimal University, Bhopal. bacterial load. Dutka (1973), Evison and Arora, D.R. 2003. Text book of James (1973), Burrel and Roland (1979), microbiology, 2ndedn. CBS publishers Hazen (1988), Ramteke et al. (1992) and distributors, New Delhi. Pp. 653 advocate that, total coliform count 659. performed by the most probable number Asthana, R.K., Singh, K.N. 1993. Physico- (MPN) method is commonly used indicator chemical characterization of Gomathi of potability despite the drawbacks that have River. Oriental J. Chem., 9(2):155 157. been well documented.

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