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 Nature Environment and Pollution Technology
Vol. 10, No. (3), 2011

CONTENTS
1. N. H. Brahmbhatt and Rinku V. Patel, Biochemical changes during lead (Pb) uptake by the alga
Pithophora 327-330
2. Shengnan Zhu, Yawen Wu, Guangjun Wen, Weirong Bai, Zhongshi Hao and Huyin Huai, Effects
of main chemical compounds in cooking oil fume condensates (COFCs) on growth of Salvinia natans
(L.) All.: II. Hexadecane 331-336
3. Mehrdad Modaresi, Effect of soybean on levels of LH, FSH and testosterone hormones and testis in adult
male mice 337-342
4. T. Padmavati, S. N. Padhy and K. C. Sahu, Studies on distribution of calcium and magnesium in
coastal waters of Gopalpur, bay of Bengal 343-350
5. B. Digamber Rao, G. Shamitha, G. Renuka and M. Ramesh Babu, Action of C-phycocyanin
pigment and cell extracts of Tolypothrix sps. on the biochemical activity of eri silkworm and their
antifungal activity 351-356
6. Naresh V. Butani, Nilesh D. Pandya, Priti R. Parmar and Piyush V. Desai, Bacterial decolourisation
of Diazo Monochlorotriazine fibre reactive dyes under optimized physico-chemical conditions 357-360
7. Fateneh Bajoul Kakahi, Satish Kulkarni and G.R. Pathade, A Review on the role of microorganisms in
treatment of paper and pulp industry effluent 361-368
8. Varsha K. Vaidya and Sharmila Kadam, Optimization of conditions for naphthalene biodegradation 369-376
9. Kailas F. Khairnar, S. T. Ingle, Nilesh D. Wagh and Lalit T. Ingale, Monitoring of railway traffic
pollution and health effects on exposed population 377-384
10. Praveen Tamot, Sadhna Tamot and Ashu Awasthi, Impact of three different types of aeration units on
limnology and fish fauna in a eutrophic tropical wetland, lower lake, Bhopal, India 385-388
11. Lakshman Nayak and Pragnya Padhi, Conservation and management of salt water crocodile (Crocody-
lus porosus) in relation to some physico-chemical parameters from Bhitarkanika Sanctuary, Orissa 389-394
12. J. Jean Jose, P. Udayakumar, M. P. Deepak, B. R. Rajesh, K. Narendra Babu and A. Chandran,
Assemblages of marine polychaete genus Glycera (Phyllodocida: Glyceridae) along the Kerala coast as an
indicator of organic enrichment 395-398
13. Nirmal Kumar, J. I., Yamini Verma, Rita N. Kumar and Manishita Das, Influence of water quality on
composition and seasonal abundance of phytoplankton community in Thol Wetland, Gujarat, India 399-404
14. M. K. Jain, L. K. Dadhich and S. Kalpana, Water quality assessment of Kishanpura Dam, Baran,
Rajasthan, India 405-408
15. M. P. Subin and V.A. Aneesha, An evaluation of physico-chemical properties to asses well water quality
in some areas of Ernakulam district, Kerala, India 409-413
16. Pavithra S. Reddy and N. Nandini , Leachate characterization and assessment of groundwater pollution
near municipal solid waste landfill site 415-418
17. Harminder Kaur, Radhe Shyam and R. Amutha, Kigelia africana fruit carbon as a low cost adsorbent for
removal of copper(II) ions from aqueous solution 419-422
18. L. P. Rema, Effect of zinc and mercury on lysosomal membrane stability of the fish Oreochromis
mossambicus (peters) 423-426
19. Meena Thomas Irimpan, C. I. Jolly and D. Sheela, Study of the preliminary phytochemistry,
antibacterial and antioxidant activities of Gymnema sylvestre R. Br. 427-429
20. Biswajit Ruj, Characteristics of hazards of coke-oven gas and blast furnace gas - A case study 431-434
21. Nidhi Poonia and M. K. Jat, Characterization and treatment of grey water for recycling 435-438
22. Ratan Chand Sharma and Shivani Dogra, Characterization of the soils of lower Himalayas of Himachal
Pradesh, India 439-446
23. Priya K. L., Gabriela Jennifer, G. Lizia Thankam, Sophia Abraham Thankam and Mariam Mathew,
Monitoring the pollution intensity of wetlands of Coimbatore, Tamil Nadu, India 447-454
24. Mohammed Naji Taresh Ali, Hina Kousar and Adamsab M. Patel, Fluoride concentration in
groundwater of Arsikere Taluk, Hassan District, Karnataka, India 455-457
25. T. G. Nagaraja, Isolation and identification of seasonal endomycophytes of inner bark of Pachira insignis 459-461
26. Snehal G. Pawar, Lalit V. Pingale, Girish R. Pathade and Vinay B. Rale, Spent wash: concomitant
pollution abatement potential employing the fungus Aureobasidium pullulans 463-465
27. Mazher Sulthana, R. Vasanthi and J. N. Jayaprakash, Hydrochemistry and seasonal fluctuation of
plankton in Arasankulam Pond at Veppampattu in Thiruvallur District of Tamil Nadu, India 467-470
28. L. Jose and Chethan Kumar, Evaluation of pollution by Palmer’s algal pollution index and physico-
chemical analysis of water in four temple ponds of Mattancherry, Ernakulam, Kerala 471-472
29. Artimes Ghassemi Dehnavi, Ramin Sarikhani and D. Nagaraju, Impact of arsenic on environmental
geochemistry of south east of Kurdistan, Iran 473-480
30. Ch. Maruthi Devi and T. Usha Madhuri, Study on groundwater quality in Prakasam District and its
suitability for drinking 481-483
31. D. N. Khairnar, A. S. Kelhe and A. B. Khairnar, Fungal diversity and mycotoxin effect on seed-borne
fungi, seed germination and seedling vigour of some cereals of Nashik District 485-486
32. Siddiqui Tasneem, Biodiversity of Ichthyofauna of Barul reservoir, Nanded District, Maharashtra 487-488
33. R. P. Mali and Shaikh Afsar, Effect of zinc sulphate on cardiac physiology of the crab Barytelphusa
guerini 489-490
34. Ayan Chandra, Gaurav Saha, Sayantan Goswami, Anwesha Saha, Soumi Sinha Roy, Payel Ganguly,
Avishek Ghosh, Kaveri Banerjee, Radhika Saraf, Arpita Banerjee and Arup Kr. Mitra, Effectivity of
vehicular banning and its reflection in disease incidence in Kolkata 491-496
35. Conferences/Symposia 414, 430
36. Environment News 458, 462, 466
37. Innovations, New Products/Processes 484
 Paryavaran Abstract, New Delhi, India

Chemical Abstracts, U.S.A.

Environment Abstract, U.S.A.

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Abstracts and full papers are available on the
LQ Journal’s Website: www.neptjournal.com

Listed in master list of journals abstracted by Thomson

Reuters

Elsevier Bibliographic Databases like Compendex, Geobase,

EnCompass, etc.

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Managaing Editor at Jaipur: Dr. Subhashini Sharma, Department of Zoology, Rajasthan University, Jaipur,
Rajastahn, India
Business Manager: Mrs. Tara P. Goel, Technoscience Publications, 2 Shila Apartment, Shila Nagar, Post Box
No. 10, Karad-415 110, Maharashtra, India

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1. Dr. Prof. Malay Chudhary, Department of Civil Engineering, 21. Dr. Prof. (Mrs.) Madhoolika Agarwal, Dept. of Botany,
Universiti Teknologi PETRONAS, Malaysia B.H.U., Varanasi, India
2. Dr. Saikat Kumar Basu, University of Lethbridge, 22. Dr. Prof. M. H. Fulekar, Deptt. of Life Sciences, University
Lethbridge AB, Canada of Mumbai, Mumbai, India
3. Dr. Sudip Datta Banik, de Instituto Politecnica Nacional 23. Dr. Prof. A.M. Deshmukh, Dept. of Microbiology, Dr. B.A.
(Cinevestav), Mexico Marathwada University Sub-Centre, Osmanabad, India
4. Dr. Elsayed Elsayed Hafez, Deptt. of of Molecular Plant 24. Dr. Prof. M.P. Sinha, Dept. of Zoology, Ranchi University,
Pathology, Arid Land Institute, Egypt Ranchi, India
5. Dr. Dilip Nandwani, CREES, Northern Marianas College, 25. Dr. Dr. G.R. Pathade, Dept. of Biotechnology, Fergusson
Northern Marina Islands, USA College, Pune, Maharashtra, India
6. Dr. Ibrahim Umaru, Department of Economics, Nasarawa 26. Dr. Dr. Ashutosh Gautam, India Glycols Ltd., Kashipur (U.P.),
State University, Keffi, Nigeria India
7. Dr. Prof. D.S. Mitchell, Albury, Australia 27. Dr. Dr. T.S. Anirudhan, Dept. of Chemistry, University of
8. Dr. Prof. Alan Heritage, Sydney, Australia Kerala, Trivandrum, Kerala, India
9. Mr. Shun-Chung Lee, Deptt. of Resources Engineering, 28. Dr. Ram Chandra, Industrial Toxicological Research
National Cheng Kung University, Tainan City, Taiwan Centre, Lucknow, India
10. Samir Kumar Khanal, Deptt. of Molecular Biosciences & 29. Dr. M.G. Bodhankar, Dept. of Microbiology, Yashwantrao
Bioengineering,University of Hawaii , Honolulu, Hawaii Mohite College, Pune, India
11. Dr. Prof. P.K. Bhattacharya, Dept. of Chemical Engineer- 30. Dr. K. Ahmed, Assam Agriculture University, Khanapara,
ing, IIT, Kanpur, U.P., India Guwahati, Assam, India
12. Dr. Prof. D.V.S. Murthy, Dept. of Chemical Engineering, IIT, 31. Dr. Biswajit Ruj, Dept.of Chemistry, C.M.E.R.I., Durgapur,
Chennai, India West Bengal, India
13. Dr. Prof. S.V.S. Chauhan, Dept. of Botany, Dr. B.R. Ambedkar 32. Dr. Sandeep Y. Bodkhe, NEERI, Nagpur, India
University, Agra, India 33. Dr. D. R. Khanna, Gurukul Kangri Vishwavidyalaya, Hardwar,
14. Dr. Prof. Arvind Kumar, Vice Chancellor, Vinoba Bhave India
University, Hazaribagh, Jharkhand, India 34. Dr. S. Dawood Sharief, Dept. of Zoology, The New College,
15. Dr. Prof. Shashi Kant, Dept. of Botany, Jammu University, Chennai, T. N., India
Jammu, India 35. Dr. B. N. Pandey, Dept. of Zoology, Purnia College, Purnia,
16. Dr. Prof. A.B. Gupta, Dept. of Civil Engineering, MREC, Bihar, India
Jaipur, India 36. Dr. B. S. Das, Indian Institute of Technology, Kharagpur, West
17. Dr. Prof. K.C. Sharma, Dept. of Environmental Science, Bengal, India
M.D.S. University, Ajmer, India 37. Dr. Ms. Shaheen Taj, Dept. of Chemistry, Al-Ameen Arts,
18. Dr. Prof. D.N. Saksena, Dept. of Zoology, Jiwaji University, Science & Commerce College, Bangalore, India
Gwalior, India 38. Dr. Nirmal Kumar, J. I., ISTAR, Vallabh Vidyanagar, Gujarat,
19. Dr. Prof. S. Krishnamoorthy, National Institute of Technol- India
ogy, Tiruchirapally, India 39. Samir Kumar Khanal, Dept. of Molecular Biosciences &
20. Dr. Prof. M. Vikram Reddy, School of Ecology & Environmenal Bioengineering,University of Hawaii, Honolulu, Hawaii
Sciences, Pondicherry University, Pondicherry, India
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N. H. Brahmbhatt and Rinku V. Patel
V. P. & R. P. T. P. Science College, S. P. University, Vallabh Vidyanagar-388 120, Gujarat, India

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,1752'8&7,21 chemical assessment of nitrogen fixing cyanobacteria,

In nature, metals constitute one of the major environmental
pollutants. Pollution by heavy metals in industrial waste ef-
fluents is now a global problem. Long term persistence of
heavy metals in the environment represents a hazardous prob-
lem leading to harmful effects on aquatic as well as human life.
Hashimoto & Furukawa (1985) indicated that the pres-
ence of algae in various effluents helped in treatment of such
wastewaters. Such information prompted us to use algae for
remediation of heavy metals from aquatic bodies. There are
several reports of algae accumulating heavy metals in their
tissues when grown in polluted waters. Through bio-
accumulation process, the concentration of the metals get
several folds higher in the organisms than their surround-
ings, which is called biomagnification. Other reports indi-
cated that carboxyl group on algal cell walls may be respon-
sible for a great portion of metal binding to inactivate algal
biomass (Gardea et al. 1990). In live algae, intracellular phos-
phate has been found to be responsible for metal sequestra-
tion (Zhang & Majidi 1994).
Pithophora is related to genus Cladophora. These algae
were found to clog waterways and lakes with their large mats
and filaments. These macro algae are also formed with the
removal of nutrients and soluble heavy metals as they form
large masses in ponds thereby providing large surface area
for binding metals. Pithophora was known to accumulate
large amounts of arsenic, lead and zinc (Imamul Huq et al.
2004).
Effect of light stress on peroxidase, succinate dehy-
drogenase and total chlorophyll content in Andrographis
paniculata has been noted by Kumar et al. (2009). Bio-
Stigonema ocellatum to acute toxicity of cadmium nitrate
was made by Kumar et al. (2009). Acute toxicity of lead
nitrate on biochemical changes of nitrogen fixing
cyanobacteria, Aulosira fertilissima has also been stud-
ied by Kumar et al. (2009). The present paper deals with
the biochemical changes in the alga Pithophora during
uptake of the heavy metal lead.
0$7(5,$/6$1'0(7+2'6
In the present work, selected algal species Pithophora sp.
for the metal removal was used. The study aimed at the ac-
cumulation of various concentrations of lead (Pb) at differ-
ent intervals of time by the algae. In in vivo experiment, short
term toxicity test was designed as per the standard methods
(APHA-AWWA-AWPC 1989). In this test to culture the
algae, metal accumulation and estimation of biochemical pa-
rameters were made.
Culture of algae: The wastewater alga Pithophora sp. was
collected from Ankleshwar GIDC area. Algae were thor-
oughly washed by tap water to remove any epiphytic algae
attached to it. The media preparation and the culturing meth-
ods were carried out following standard methods (APHA-
AWWA-AWPC 1989). Duration of incubation period was
3, 6 and 9 days.
Metal accumulation: The stock solution of 1000 ppm of
lead was prepared in double distilled water by dissolving
lead acetate (C2H3O2)2Pb.3H2O. For the experiment, 2, 5, 10,
20 and 30 ppm concentration of the metal was used. Algal
samples, harvested after 3, 6 and 9 days of incubation, were
washed repeatedly, dried and digested in HNO3/HClO4 mix-
ture (4:1, v/v) for heavy metal analysis. Estimation of lead
 N. H. Brahmbhatt and Rinku V. Patel

was made on ICP [Inductively Couple Plasma Spectrometer,

Perkin Elmer Corporation (ICP Optima 3300RL)].
Estimation of biochemical parameters: Peroxidase was de-
termined following Sadasivam & Manickam (1992) and
sugar was determined by GOD-POD test. Protein content
was determined in accordance with the method of Lowry et
al. (1951). Catalase and proline concentrations were deter-
mined following Thimmaiah (1999). Chlorophylls a,b and
total chlorophyll concentrations were determined by the pro- )LJ&RPSDULVRQRIDFFXPXODWLRQRI3ELRQDWGLIIHUHQW
GXUDWLRQVRILQFXEDWLRQSHULRGLQ3LWKRSKRUD
cedures of Arnon (1949).
In the present study, the activity of catalase decreased in
5(68/76$1'',6&866,21 Pb ion treatments after each day of exposure in Pithophora
Removal of harmful metals from the environment, particu- (Table 3). The increased exposure periods lower the enzyme
larly from the aquatic water bodies as well as from the in- activity because of loss of pigment, protein and proline in-
dustrial wastes is absolutely essential before they enter into duced due to metal ions interfering with the physiological
food chains. Algae have low and simple nutrient require- activity of the cells.
ments, grow rapidly and easily thrive in aquatic environ- Earlier, it was reported that exposure of several types of
ment and can be well studied physiologically as compared stresses viz., heat, salinity, osmotic stress, metal exposure,
to higher plants. Algae produce oxygen for other aquatic biota results in alterations in protein synthesis. In the present study
and oxidise organic materials present in wastes making
wastewater more favourable for release into the environment.
There are no reports of dead higher plant materials remov-
ing metals, but algae even when dead, efficiently remove
metal ions (Morendo-Garrido et al. 1998). The alga selected
in the present study is used with a view to provide a useful
solution of problems of metal pollution.
Somashekar (1982) reported Pithophora as one of the
dominant algae in the effluents from electroplating industry
Fig. 2: Pithophora after accumula- Fig. 3: Pithophora after accumu-
where it could accumulate 15mg/L of metals. It was also tion of 2 ppm Pb on 6 day. lation of 5 ppm Pb on 6 day.
investigated in the present work that Pithophora accumu-
lated higher level of lead. The differential absorption
capabilities of the metal ions in different algal groups de-
pend on several other external factors such as pH, background
metal ion concentration, light intensity, chemical make up
of the medium as well as growth phase and cell division
stages (Skipnes et al. 1975). Pithophora showed more affin-
ity towards Pb ions and this might be the reason that lead is
accumulated in higher amounts in the cells. In present work
accumulation of Pb ions was higher with 8.96 mg/L at 5 Fig. 4: Pithophora after accumula- Fig. 5: Pithophora after accumula-
ppm concentration of 6 days and lower with 0.28 mg/L at 10 tion of 10 ppm Pb on 6 day. tion of 20 ppm Pb on 6 day.
ppm concentration of 6 days (Table 1, Fig. 1).
In Pithophora, peroxidase activity increases after 3 days
of exposure to Pb (Table 2). There are reports suggesting the
increase in peroxidase activity under metal ion stress.
Mukherji & Maitra (1976) demonstrated the stimulated ac-
tivities of catalase, peroxidase and IAA oxidase under con-
ditions of Pb toxicity in rice seedlings. The increased per-
oxidase activity is a response to an increase in peroxides,
which reduce disruption of plasma membrane by lipid
peroxidation. Fig. 6: Pithophora after accumulation of 30 ppm Pb on 6 day.

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 BIOCHEMICAL CHANGES DURING LEAD UPTAKE BY PITHOPHORA 

Table 1: Uptake of Pb ions at various concentrations (mg/L) after 3, 6 and Table 2: Estimation of peroxidase (mg/g) at various Pb concentrations in
9 days of exposure by Pithophora sp. Pithophora sp.

No. Concentration (Pb) 3 Days 6 Days 9 Days No. Concentration (Pb) 3 Days 6 Days 9 Days

1 Control 0.1352 0.1352 0.1352 1 Control 0.890 0.890 0.889

2 2 ppm 0.6685 4.2451 1.4714 2 2 ppm 0.891 0.739 0.729
3 5 ppm 1.9631 8.9600 2.1775 3 5 ppm 0.893 0.740 0.735
4 10 ppm 4.5061 0.2808 3.2154 4 10 ppm 0.898 0.746 0.731
5 20 ppm 2.2129 7.7945 4.4331 5 20 ppm 0.900 0.751 0.743
6 30 ppm 6.9655 6.5344 8.9289 6 30 ppm 0.911 0.779 0.771

Table 3: Estimation of catalase (mg/g) at various Pb concentrations in Table 4: Estimation of protein (mg/g) at various Pb concentrations in
Pithophora sp. Pithophora sp.

No. Concentration (Pb) 3 Days 6 Days 9 Days No. Concentration (Pb) 3 Days 6 Days 9 Days

1 Control 25.4 25.3 25.1 1 Control 10.92 10.61 10.56

2 2 ppm 27.0 23.2 22.9 2 2 ppm 10.87 10.76 10.67
3 5 ppm 26.5 21.7 21.1 3 5 ppm 11.56 11.02 10.88
4 10 ppm 25.0 20.7 20.3 4 10 ppm 11.03 10.56 10.35
5 20 ppm 24.2 20.2 19.5 5 20 ppm 10.69 10.07 9.01
6 30 ppm 23.0 20.0 19.1 6 30 ppm 10.11 9.80 9.00

Table 5: Estimation of proline (mg/g) at various Pb concentrations in Table 6: Estimation of sugar (mg/g) at various Pb concentrations in
Pithophora sp. Pithophora sp.

No. Concentration (Pb) 3 Days 6 Days 9 Days No. Concentration (Pb) 3 Days 6 Days 9 Days

1 Control 0.883 0.880 0.877 1 Control 20.7 20.7 20.7

2 2 ppm 0.863 0.861 0.857 2 2 ppm 24.5 23.2 22.5
3 5 ppm 0.861 0.851 0.846 3 5 ppm 23.0 22.8 21.7
4 10 ppm 0.858 0.822 0.820 4 10 ppm 24.0 23.6 23.8
5 20 ppm 0.843 0.819 0.810 5 20 ppm 20.2 21.4 22.1
6 30 ppm 0.839 0.811 0.808 6 30 ppm 19.2 19.9 20.9

Table 7: Estimation (mg/g) of chlorophyll a, b and total chlorophyll at various Pb concentrations in Pithophora sp.

No. Concentration 3 days 6 days 9 Days

(Pb) Chl-a Chl-b Total chl Chl-a Chl-b Total chl Chl-a Chl-b Total chl

1 Control 0.217 0.158 0.375 0.216 0.158 0.374 0.216 0.157 0.373
2 2 ppm 0.217 0.165 0.382 0.213 0.162 0.375 0.212 0.158 0.370
3 5 ppm 0.211 0.167 0.378 0.208 0.159 0.367 0.202 0.159 0.361
4 10 ppm 0.212 0.147 0.359 0.205 0.145 0.350 0.201 0.144 0.345
5 20 ppm 0.216 0.161 0.377 0.212 0.160 0.372 0.210 0.146 0.356
6 30 ppm 0.213 0.151 0.364 0.207 0.149 0.356 0.205 0.145 0.350

it was observed that the algae showed enhanced protein level
after first exposure period i.e., after 3 days at 5 ppm concen-
tration, which is higher than the others (Table 4). Enany &
Issa (2001) investigated the accumulation of proline and cor-
related it with protein content. Pithophora showed reduc-
tion in the amount of protein at 20 ppm and 30 ppm Pb level.
This might be due to degradation of protein at extreme con-
centration of the metal ions. The degradation of protein con-
tent was reported after 6 and 9 days in Pithophora.
It was found that the proline content decreases day by
day after the treatment with Pithophora. After treatment pro-
line accumulation has highest value of 0.863 mg/g at 3 days
at 2 ppm concentration and lower value of 0.808 mg/g after
9 days at 30 ppm exposure (Table 5). As a result of less pro-
line amount after the metal exposures, the cells have lower
survival and less protected.
The sugar content has decreased almost continuously in
Pithophora except on 3 days at 2 ppm, when it was higher

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 N. H. Brahmbhatt and Rinku V. Patel
than others. The increased concentration of metal ion has
resulted in lower sugar content of 19.2 mg/g at 30 ppm after
3 days of treatment in Pithophora (Table 6).
The chlorophyll pigment of the algae Pithophora under
study was affected by heavy metal exposure (Table 7). Inhi-
bition in chlorophyll contents in the algae supports the ear-
lier findings, which suggested that the impaired metabolic
status imposed by toxic metals, might be associated with
either induction or activation of specific degrading enzymes
leading to the breakdown of cell organelles and macromol-
ecules (Nag et al. 1984).
&21&/86,21
The study proves that algae can sequester metal ion in their
cells with differential uptake capacity towards absorption
of different metals. As metals hinder normal biological proc-
esses in algae, they affect various biochemical parameters
like enzymes catalase and peroxidase, protein, proline, sugar
and chlorophylls in the algae . As a response to heavy metal
stress, proline accumulation was found in the algae. Pro-
longed exposure to metal ions caused inhibitory effect on
the synthesis of chlorophylls, protein and proline contents
of the algae. Peroxidase activity, in general, exhibited a rise
in Pithophora, while the activity of catalase decreased. There-
fore the survival of the algae under heavy metal stress might
be due to increased activity of peroxidase and not due to
catalase.
$&.12:/('*(0(17
The authors are grateful to UGC, New Delhi for providing
financial assistance to carry out the investigations under the
special assistance programmed of S.P. University, Gujarat.
5()(5(1&(6
APHA-AWWA-WPCF 1989. Standard Methods for the Examination of
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,1752'8&7,21 However, we have little knowledge on what components in

Water pollution, especially of surface water, is becoming a
serious environmental problem worldwide (Lemly 2004, Li
et al. 2007, Oberholster 2008, Yoshiaki 2008). There are
many different pollutants contaminating water environments
(Goel 2006) such as heavy metals (Dixit & Tiwari 2008,
Kar et al. 2008), industrial and agricultural wastes (Ramos
et al. 2006), etc. Water pollution can impact economic de-
velopment (Reddy & Behera 2006), living standards (Dwight
et al. 2005, Li et al. 2007), public health (Ake & Bo 2005),
etc. Among the pollutants, oil and grease from food manu-
facturing industries (such as dairies and slaughter houses)
and domestic activities is a serious one (Omil et al. 2003,
Khan et al. 2004, Cammarota & Freire 2006). It has many
ecological effects (Salanitro 1997, Jiang et al. 2009). Cook-
ing oil fume condensates (COFCs), a kind of waste dis-
charged from restaurants and domestic cooking, are one of
the sources of oil and grease pollutants (Ko et al. 2000,
Metayer et al. 2002, Miao et al. 2005, Jiang et al. 2009). It is
obvious that the amount of COFCs will increase with ur-
banization. It is reported that 4600 restaurants in Guangyuan
city will collect at least 460 kg/d COFCs (Yan et al. 2003).
There is no effective method for treating COFCs presently.
Much of it is discharged directly as rubbish. COFCs have a
significant effect on the vegetative growth of Salvinia natans,
a floating-leaves aquatic plant (Jiang et al. 2009). There are
over 100 components in COFCs, such as polycyclic aromatic
hydrocarbons, hexanal and 2-heptenal, aromatic amines,
alkyl, alkene, and aldehyde (Leson & Winer 1991, Chiang
et al. 1999, Zhu & Wang 2003, Kawai et al. 2006).
COFCs contribute to its ecological effects. Hydrocarbons
are very important chemical components in COFCs because
of their great amount and diversity (Liu et al. 2002). In our
earlier paper, we have reported that dodecane, one of hydro-
carbons in COFCs, has significant influences on the vegeta-
tive growth of S. natans (Wu et al. 2011). We discuss the
effects of another hydrocarbon, hexadecane, on the growth
of S. natans, and compare the influences on growth of float-
ing aquatic plants between dodecane and hexadecane in this
paper.
0$7(5,$/6$1'0(7+2'6
Collection of S. natans and treatment by hexadecane: The
collection and adaptation of culture of S. natans are con-
ducted as described in our earlier paper (Wu et al. 2011).
Static toxicity testing method has been used. Hexadecane
was diluted in distilled water to prepare test solutions with
the following concentration gradient: 0 mg/L (Control), 10
mg/L (C1), 20 mg/L (C2), 40 mg/L (C3), 80 mg/L (C4), 160
mg/L (C5), 240 mg/L (C6), and 320 mg/L (C7).
One hundred and sixty healthy individuals of S. natans
with similar size (four pairs of the latest floating leaves-old
leaves, similar lengths and fresh weights) were assigned ran-
domly to 8 groups (one control and seven treatments) and
treated by test solutions according to the method in our ear-
lier reports (Jiang et al. 2009, Wu et al. 2011). Each indi-
vidual of S. natans had only one apical bud and no branch at
the beginning of the experiment. A plastic pot (Ø12 cm and
10 cm high) contained one individual only and the pots were
 Shengnan Zhu et al.
kept in a greenhouse (at temperature 30±2°C, the light was
provided by metal halide bulbs for 12 h/d).
The following parameters were recorded on day 2, 4, 6,
9 and 12: mortality, number of leaves, number of buds, stem
length, number of leaves turning yellow and conductivity of
the test solution. The pH value was also measured on day 0,
2, 4, 6, 9 and 12. The leaf area, fresh weight and dry weight
of each individual were measured at the end of the experi-
ment.
The analysis of data: The relative growth rate (RGR) and
percent inhibition of growth rate (% I) were determined based
on the methods suggested by OECD (2006), the two equa-
tions have been introduced in our earlier papers (Jiang et al.
2009, Wu et al. 2011). In this paper,'t =12 days.
One-way analysis of variance (ANOVA) was used to de-
termine the differences among different treatments, and lin-
ear regression was used for analysis of correlation between
the parameters of the vegetative growth of S. natans and the
concentration of hexadecane with SPSS 16.0. All variables
were tested for normality and homogeneity of variances. The
differences were statistically significant at p<0.05.
5(68/76
Effects of hexadecane on RGR and % I of S. natans: The
RGR in different groups are shown in Fig. 1. The RGR was
much higher in control than in C4-C7 (df = 68, F = 6.421,
p<0.001), but there was no significant difference between
control and C1-C3 (df = 77, F = 0.577, p = 0.632). There was
a significant linear correlation between % I and the concen-
tration of hexadecane (R2 = 0.949, F = 93.501, p<0.001) (Fig.
2). The results indicated that hexadecane limited the RGR of
S. natans significantly, and its effect was correlated posi-
tively to its concentration, especially when > 40mg/L.
Effects of hexadecane on the leaves of S. natans: Although
the leaf number of S. natans in all experimental groups in-
creased stably during the experiment, C5-C7 increased more
slowly than other groups after day 4 (Table 1). As the same
as the variation of leaf number, the total leaf area of control
was significantly higher than that of C5-C7 (df = 49, F =
33.181, p<0.001). When the concentration of hexadecane was
up to 80mg/L, it had no significant effect on the leaf area of
S. natans (df = 96, F = 0.465, p = 0.761) (Fig. 3).
Hexadecane could accelerate the leaves of S. natans turn-
ing yellow, and its effect was correlated closely with its con-
centration and the exposure time (Fig. 4). The number of old
leaves turning yellow in C1-C7 was significantly higher than
that in control after day 6 (df = 23, F = 21.158, p<0.001).
The old leaves in C6-C7 completely turned yellow after day
9 (Fig. 4). However, the effect of hexadecane on the new
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
leaves was not as acute as the old leaves. There were no new
leaves turning yellow in control during the experiment, but
in all of the treatment groups until day 12. There were sig-
nificant differences between control and C4-C7 while no
difference between control and C1-C3 from day 4. The in-
fluence of hexadecane on the old leaves of S. natans was
more significant than the new leaves.
Effects of hexadecane on the stem and the bud of S.
natans: The stem length of S. natans in all experimental
groups went up during the experiment (Fig. 5). The stem
length in control was significantly higher than that in C6-
C7 (df = 14, F = 8.855, p = 0.004). However, there was no
significant difference between control and C1-C5 (df = 29,
F = 0.37, p = 0.864). The result indicated that hexadecane
would significantly limit the growth of the stems of S. natans
when its concentration was more than 160mg/L.
The accumulated number of buds in control and C1-C4
increased stably during the experiment (Table 2). At the end
of the experiment, the accumulated number of buds in con-
trol was significantly higher than that in C3 and C5-C7.
Hexadecane could influence development of the buds of
S. natans. The higher the concentration is, the more signifi-
cant effect on the growth of buds would be.
Effects of hexadecane on the root of S. natans: The sub-
merged leaves of S. natans have the function of absorbing
nutrients from water like ‘root’. The plants produced fewer
‘roots’ per frond when exposed to hexadecane, and the length
of ‘roots’ was also affected significantly by hexadecane in
C3 and C5-C7 during the experiment (Fig. 6).
Effects of hexadecane on the biomass of S. natans: The
dry weights of different organs of S. natans at the end of the
experiment are showed in Fig. 7. The dry weight of floating
leaves (df = 126, F = 5.107, p<0.001), stems (df = 126, F =
7.335, p<0.001) and roots (df = 126, F = 5.541, p<0.001) in
control were significantly higher than those in the treatment
groups. Except the root, the dry weight of floating leaves,
stems and whole plants were correlated negatively with the
concentrations of hexadecane (Table 3). The results showed
that hexadecane would influence the accumulation of
biomass of S. natans. The higher the concentration of
hexadecane is, the lower biomass of S. natans would be.
Effects of hexadecane on the mortality of S. natans: Only
in control and C2, no dead individuals of S. natans have
been observed during the experiment. At the end of the ex-
periment, there were only 7 individuals (35%) survived in
both C6 and C7. In these two groups, the dead individuals
were observed on day 2 firstly (Table 4). Thirty three indi-
viduals died during the experiment totally, and 24 (72.7%)
died until day 4. There was significantly positive linear cor-
 EFFECTS OF HEXADECANE ON GROWTH OF SALVINIA NATANS 

Table 1: The number of leaves of S. natans in different experimental groups during the experiment (m ± SE, n = 20).

Experimental group Day 2 Day 4 Day 6 Day 9 Day 12

Control 11.0±0.3 16.1±0.7 24.9±0.9 38.7±1.9 54.6±3.2

C1 11.2±0.3ns 16.7±0.6ns 24.4±1.1ns 36.6±2.1ns 51.2±2.4ns
C2 11.1±0.2ns 16.1±0.5ns 22.3±1.0ns 33.0±1.7* 48.4±2.7ns
C3 11.2±0.3ns 16.6±0.7ns 23.7±1.2ns 35.5±2.2ns 45.4±2.8*
C4 11.8±0.3ns 17.8±0.8* 26.2±1.5ns 38.2±2.0ns 48.5±3.0ns
C5 10.5±0.2ns 13.8±0.5* 18.1±0.9** 23.4±1.7** 27.8±2.2**
C6 10.2±0.3ns 13.1±0.7** 15.3±1.3** 19.4±2.4** 21.7±3.4**
C7 10.3±0.3ns 13.2±0.7* 15.6±0.7** 17.1±0.9** 18.9±1.1**

‘ns’ indicates that there are no differences between the control and the experimental group (p>0.05); * indicates that there are differences between the
control and the experimental group (p<0.05); ** indicates that there are significant differences between the control and the experimental group (p < 0.01).

Table 2: Mean number of buds of S. natans in different experimental groups during the experiment (m ± SE, n = 20).

Experimental group Day 2 Day 4 Day 6 Day 9 Day 12

Control 1.9±0.2 3.7±0.3 4.6±0.3 5.4±0.4 6.5±0.5

C1 2.4±0.3ns 3.3±0.3ns 3.8±0.3* 4.5±0.4ns 6.0±0.5ns
C2 1.7±0.2ns 3.4±0.3ns 3.6±0.3* 4.9±0.4ns 6.2±0.5ns
C3 2.4±0.2ns 2.6±0.3** 3.2±0.3** 3.7±0.3** 4.7±0.4**
C4 1.9±0.2ns 2.8±0.2* 3.6±0.3** 4.6±0.5ns 5.5±0.5ns
C5 1.8±0.2ns 2.3±0.2** 2.5±0.2** 3.1±0.3** 3.1±0.3**
C6 1.1±0.2* 1.7±0.2** 1.8±0.2** 1.7±0.2** 1.2±0.2**
C7 1.1±0.2* 1.9±0.1** 1.9±0.1** 1.9±0.1** 2.0±0**

‘ns’ indicates that there are no differences between the control and the experimental group (p>0.05); * indicates that there are differences between the
control and the experimental group (p<0.05); ** indicates that there are significant differences between the control and the experimental group (p < 0.01).

Table 3: The regression equations between the dry weight of different The pH value and conductivity of the cultivation media
organs (y) and the concentration of hexadecane (x). during the experiment: The pH value and conductivity of
Organ Regression equation R2 p
the cultivation media in different experimental groups had
similar trends with the treatment time (Fig. 9). There was no
Floating leaves y = -0.000021x + 0.028 0.605 0.023 difference of pH values (df = 47, F = 0.029, p = 1.000) and
Stems y = -0.0000078x + 0.005 0.654 0.015 conductivities (df = 47, F = 1.831, p = 0.108) among the
Roots y = -0.0000065x + 0.007 0.289 0.17
Whole plants y = -0.000035x + 0.04 0.608 0.023
experimental groups during the experiment. This result sug-
gested that the presence of hexadecane had no effect on the
Table 4: Mortality of S. natans in different experimental groups from day 2 pH value and conductivity of the water environment.
to day 12.
',6&866,21
Experimental Day 2 Day 4 Day 6 Day 9 Day 12
group Hydrocarbons are considered as the most threatening haz-
ardous pollutants from oils due to their persistence in envi-
Control 0% 0% 0% 0% 0% ronments as well as their mutagenic and carcinogenic prop-
C1 0% 0% 0% 5% 5% erties (Lemiere et al. 2005). Although hexadecane and
C2 0% 0% 0% 0% 0%
C3 0% 0% 0% 5% 5% dodecane both belong to hydrocarbons, they show a little
C4 0% 0% 0% 0% 5% different influence on the growth of S. natans. LC50 of
C5 0% 10% 10% 15% 20% hexadecane (275 mg/L) to S. natans on day 4 after treatment
C6 25% 55% 60% 65% 65% is between that of COFCs (801 mg/L) and dodecane (190
C7 15% 55% 60% 65% 65%
mg/L) (Jiang et al. 2009, Wu et al. 2011). This means that
relation between the mortality and the concentration of hexadecane is another important component in COFCs,
hexadecane (Fig. 8). LC50 of hexadecane for S. natans based which contributes greatly to the ecological effects of COFCs.
on probit analysis was 275 mg/L on day 4 and 244 mg/L on It can limit the bud, stem and leaf growth of S. natans. It
day 12, respectively. also can accelerate the leaves becoming yellow, then influ-

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Fig. 1: The RGR (mean ± SE, n = 20) of S. natans in different experimental
groups. * indicates that there are differences between the control and the

experimental group (p<0.05). ** indicates that there are significant differ- 'D\ 'D\ 'D\ 'D\ 'D\
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ences between the control and the experimental group (p<0.01).
 




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Fig. 4: The numbers of old and new leaves turning yellow (mean ± SE, n =
Fig. 2: The relationship between % I and the hexadecane 20) in the experimental groups on day 2, 4, 6, 9, 12. * indicates that there
concentration at the end of the experiment. are differences between the control and the experimental group (p<0.05).
** indicates that there are significant differences between the control and
the experimental group (p < 0.01).


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Fig. 3: The total leaf area (mean ± SE, n = 20) of S. natans in different        
experimental groups. * indicates that there are differences between the 7KHWLPHDIWHUWUHDWPHQWG
control and the experimental group (p<0.05). ** indicates that there are
significant differences between the control and the experimental group Fig. 5: The stem length of S. natans in different experimental groups
(p<0.01). during the treatment (mean ± SE, n = 20).

ence the photosynthesis and biomass accumulation of S. hexadecane and dodecane do not change the pH value of
natans. It can lead to the death of S. natans at the early stage. cultivation media significantly. Hexadecane has no effect
S. natans is more sensitive to hexadecane than to COFCs. on the conductivity of the cultivation media during the ex-
COFCs can influence the pH value of water when it en- periment. This means that hexadecane does not influence the
ters a water body (Jiang et al. 2009). However, both vegetative growth of S. natans by changing the water envi-

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Fig. 6: The length of ‘roots’ (mean ± SE, n = 20) of S. natans in different 
experimental groups. * indicates that there are differences between the        
control and the experimental group (p<0.05). ** indicates that there are 7KHWLPHDIWHUWUHDWPHQWG
significant differences between the control and the experimental group
(p<0.01). 

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experimental groups during the treatment (mean ± SE, n = 20).
Fig. 7: The dry weights of different parts of S. natans (mean ± SE, n = 20)
of S. natans in different experimental groups. * indicates that there are dif- film on the surface of water. This oil film will cover the sur-
ferences between the control and the experimental group (p<0.05). ** in-
dicates that there are significant differences between the control and the
face of leaves, stems and buds of S. natans, then influence
experimental group (p<0.01). the photosynthesis of the plant.
For the components in oil and grease with lower relative
 density (<1.0), they will float on the surface of water when
 \ [ they enter a water body. They contact the floating plants di-



5  S  rectly, especially their leaves. This oleaginous film may in-
fluence air exchanges of leaves through stomata with the
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outside (Jiang et al. 2009). Many studies have reported that

aqueous components from crude oil can disturb stomatal
behaviour significantly (Youssef 2002), or reduce chloro-

phyll contents in leaves (Achuba 2006). The variation of sto-

       
matal behaviour and chlorophyll content will influence the
+H[DGHFDQHFRQFHQWUDWLRQPJ/ 
 photosynthesis of plants (Rzepka et al. 2005, Fernandez
2006). So oil and grease may have more serious influence
Fig. 8: The relationship between mortality and different concentrations on floating-leaves plants than other kinds of aquatic plants.
of hexadecane.
&21&/86,21
ronment. It may directly affect the plant self. It also suggests
that there are other components in COFCs which lead to As main chemical components of COFCs and other oil and
change of the pH value of the water. The molecular weight grease, both hexadecane and dodecane significantly influ-
of hexadecane is 226.44 and the relative density is 0.773. It ence the growth of leaf and bud of S. natans by disturbing
is difficult to be dissolved in water and will form an ‘oil’ its photosynthesis. S. natans can be used as an indicator of

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Shengnan Zhu et al.
hexadecane, as well as of dodecane. Hydrocarbons in oil and
grease are important chemical components which can affect
the growth of aquatic plants, especially the floating-leaves
plants. It is very necessary to study the relationship between
the amount of hydrocarbons in COFCs and the heated tem-
perature of edible oil. The low amount of hydrocarbons in
COFCs may reduce its ecological effects. Additionally, be-
cause hexadecane may influence floating-leaves plants di-
rectly, such as influence their photosynthesis and other physi-
ological processes, it is also necessary to conduct experi-
ments to test how hexadecane affects floating-leaves plants
at physiological levels.
$&.12:/('*(0(17
Mr. Wei Xu, Mr. Shaohua Sun, Mr. Jianing Xu and Mr. Jialei
Zhu participated in parts of the experiment. This study is
supported financially by the Science Creative Program of
Yangzhou University.
5()(5(1&(6
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T. Padmavati, S. N. Padhy and K. C. Sahu
Department of Marine Sciences, Berhampur University, Berhampur-760 007, Orissa, India

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,1752'8&7,21 correlation between chlorinity and calcium (P>0.01) and

Major ions are defined as those elements whose seawater
concentration is greater than 1 ppm. The main reason for
this definition to be used is because salinity is reported to ±
0.001 ppt or 1 ppm. Thus, the major ions are those ions that
contribute significantly to the salinity. According to this defi-
nition there are 11 major ions present in seawater. Amongst
all the alkaline earths occur in seawater, magnesium and cal-
cium are the major constituents, which play an important
role in biogeochemical processes in sea. The behaviour of
major elements in seawater all over the world was reported
by Marcet (1819) and they bear nearly the same proportion
to each other. Culkin (1965) and Wilson (1975) reported
that the Ca/Cl and Mg/Cl ratios of seawater are used to as-
sess the existing chemical and biological processes.
They further stated that input of calcium and magnesium
is largely independent of salinity and varies from place to
place where they accounted for the local circumstances such
as (i) abundance of life forms, (ii) presence of rivers, and
(iii) geological and meteorological conditions. The biologi-
cal processes account for the uptake of calcium and magne-
sium by organisms and their removal from seawater is very
pertinent to geochemical processes. Though, the distribu-
tion of calcium and magnesium and their ratios to chlorinity
in Indian seas have been studied by several workers (Naik
1978, Naronha et al. 1981, Sen Gupta et al. 1978), limited
information is available for Bay of Bengal. Tripathy et al.
(1990) in their studies on distribution of Ca and Mg in the
Rushikulya estuary of Orissa, observed strong positive
chlorinity and magnesium (P>0.01) confirming the conserva-
tive behaviour in a more general way.
Apart from the above, very little efforts have been made
on the distribution of calcium and magnesium in coastal and
offshore waters of this country. Due to influx of pollutants
and freshwater, the variation of the major ions as well as
salinity affect the ratio of the ions to chlorinity. Making a
study on the major ions and their ratios with chlorinity will
give an index of coastal water pollution. In view of this, the
present work was undertaken for a period of one year (March
2007-February 2008) in the near shore waters of Gopalpur
Port.
678'<$5($
The study area of Gopalpur falls on 19°6’ lat. N and 84°55’
long. E., where three transacts were selected as shown in
Fig. 1. The water samples were collected both from surface
and bottom during monsoon (June-September), post-
monsoon (October-February) and pre-monsoon (March-
May) periods. Gopalpur on the south Orissa coast is endowed
with marine resources, which fetch good financial earning
to the state. The coastal sand is famous for its rich rare earth
minerals, and there is an operational public limited company
called Orissa Sands Complex (Indian Rare Earth Limited)
to tap the resources like monazite, ilmenite, rutile, etc. There
is a caustic soda producing unit named Jayshree Chemicals
Ltd. on the bank of Rushikulya river near the estuary. This
area also houses large patches of salt producing fields at
 T. Padmavati et al.
Humma and Sorol near Rushikulya and Bahuda estuarine
areas respectively. The study area is not only manifested by
different resource types, but also important to the tourist point
of view, and occupies a prominent status on the fishing
ground of the state. Gopalpur-on sea is a major tourist resort
in the study area and its scenic beauty attracts the tourists
from all over the country. Other tourist spots include
Potagarh, an archaeological site near the Rushikulya estu-
ary; Arjipalli, the widest bathing beach; Dhabaleswar and
Mantridi, religious spots, and Sonepur for bathing beach and
scenic beauty. The northern part of the coast shows a condu-
cive environment for breeding and mass nesting of Olive
Ridley (Lepidochelys olivacea) sea turtles, which come in
thousands to lay eggs on the beach near Purunabandh vil-
lage at Rushikulya estuary. This area is increasingly getting
attention by the environmental lobbyists to declare as a ma-
rine sanctuary by the State Government. In regard to the an-
thropogenic activities, the coastal waters exhibit variability
in the physico-chemical characteristics of the water and ac-
count for a divesting effect to the marine ecosystem. The
seasonal variation of some selected major elements (calcium,
magnesium, chloride) were assessed from the coastal water
of the coast.
0$7(5,$/6$1'0(7+2'6
The surface water samples were collected in polythene
bucket, while bottom water samples were collected using
Fig. 1: Map showing the sampling locations.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
shallow water sampler. The samples were filtered through
Whatman filter paper. Before filling; the acid cleaned poly-
thene bottles were rinsed 2-3 times with respective sample
water. These filled polythene bottles with leak proof stop-
pers were preserved in a refrigerator till analysis, which was
completed within a month from the date of collection. Gen-
erally, the samples preserved in refrigerator will not allow
change in ionic concentration even after one month of storage.
Determination of chlorinity was made by Knudsen’s
titrimetric method (Grasshoff 1976). Calcium and magne-
sium were determined after titrating the samples to the pho-
tometric end point with EDTA solution. Complexometric
titration with EDTA and Eriochrome Black T indicator
yielded combined concentrations of calcium and magnesium
concentration.
5(68/76$1'',6&866,21
The seasonal variability of the water characteristics of near
shore waters depends on the freshwater discharge and asso-
ciated inflow of saline water into the area. In the present
study, the near shore waters of Gopalpur receive a signifi-
cant influx of freshwater from the River Rushikulya on its
north and the Bahuda on its south apart from other creeks
contributing to some extent. The water discharge is regu-
lated predominantly by the regime of rainfall.
Salinity: The data on level of salinity in seawater are given
 DISTRIBUTION OF CALCIUM AND MAGNESIUM IN COASTAL WATERS OF GOPALPUR
in Table 1 and Fig. 2. The surface values of salinity vary
from 30.98 to 33.15 ppt during premonsoon, 32.92 to 29.04
ppt during monsoon and 27.94 to 30.48 ppt during
postmonsoon periods at Station-1; from 31.56 to 33.18 ppt
during premonsoon, 33.10 ppt during monsoon and 28.08
to 30.52 ppt during postmonsoon at Station-2; and from
31.16 to 33.12 ppt during premonsoon, 32.52 to 28.96 ppt
during monsoon and 22.09 to 22.09 ppt during post monsoon
at Station-3. The bottom values varied from 31.04 to 33.34
ppt during premonsoon, 33.44 to 29.48 ppt during monsoon
and 28.71 to 30.95 ppt during postmonsoon at Station-1;
from 32.78 to 33.38 ppt during premonsoon, 33.22 to 30.16
ppt during monsoon and 28.98 to 30.99 ppt during
postmonsoon at Station-2; and from 31.33 to 33.32 ppt dur-
ing premonsoon, 33.13 to 29.16 ppt during monsoon and
22.38 to 29.16 ppt during post monsoon at Station-3. The
high values of salinity were recorded during the premonsoon
months and also in June. The first sign of dilution was no-
ticed during August and September , which gradually de-
creased to minimum in the month of October. But there was
a sudden increase during November.
During premonsoon period, the monthly variation of sa-
linity at surface waters is lower than the bottom waters and
also high in June due to maximum evaporation and the in-
flux of freshwater. In the month of February the water slowly
begins to recover from the dilution. On an average, the sur-
face values were lower than the bottom values because of
continuous exchange of freshwater through adjacent creek.
High salinity at Gopalpur is attributed to the onset of high
salinity associated with northward flowing coastal current.
Calcium: The data on level of calcium in seawater are given
in Table 3 and Fig. 3. The values of calcium in surface wa-
ters varied from 0.406 to 0.401g/kg during premonsoon,
0.375 to 0.4 g/kg during monsoon and 0.421 to 0.371 g/kg
at Station-1; from 0.357 to 0.369 g/kg during premonsoon,
0.375 to 0.410 g/kg during monsoon and 0.411 to 0.333 g/
kg during postmonsoon at Station-2, and from 0.403 to 0.394
g/kg during premonsoon, 0.396 to 0.350 g/kg during
monsoon and 0.381 to 0.373 g/kg during postmonsoon at
Station-3. The bottom water values ranged from 0.446 to
0.395 g/kg during premonsoon, 0.390 to 0.415 g/kg during
monsoon and 0.421 to 0.358 g/kg during postmonsoon at
Station-1, from 0.373 to 0.401 g/kg during premonsoon,
0.385 to 0.395 g/kg during monsoon and 0.415 to 0.393
g/kg during postmonsoon at Station-2, and from 0.345 to
0.393 g/kg during premonsoon, 0.396 to 0.375 g/kg during
monsoon and 0.383 to 0.429 g/kg during postmonsoon at
Station-3. The average concentration of Ca in bottom water
was higher than the surface water. Lowering of Ca in sur-
face waters may be due to the biological utilization, degra-
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

dation of dead animals, precipitation of Ca and its carbon-
ates, and settling through the particulate matter. The maxi-
mum concentration of calcium was observed during the
months of March and April. The reason may be the less bio-
logical utilization and insignificant influx of freshwater.
During August, September and October, there was high
concentration of Ca due to maximum influx of freshwater.
During postmonsoon season, the Ca concentration was com-
paratively lower. Table 2 gives values of chlorinity and
Table 4 describes the monthly calcium to chlorinity ratio at
the three stations. Calcium to chlorinity ratio in surface wa-
ters vary from 0.02009 to 0.02723 at Station-1, 0.01964 to
0.02593 at Station-2, and 0.01943 to 0.03822 at Station-3.
In the bottom waters, the ratio varies from 0.01967 to 0.02649
at Station-1, 0.01958 to 0.02611 at Station-2, and 0.01924
to 0.03327 at Station-3. During monsoon months, in most
of the cases, the ratio for surface water was higher than the
bottom water. In October month Ca/Cl ratios show higher
values for all the stations, typically confirming that the dilu-
tion effect during these months play a significant role in Ca/
Cl ratio. Station-3 values confirm this condition in a more
regular sense. The Ca/Cl ratio for the surface water is higher
than the bottom during the monsoon months and October
due to the process of dilution. Its effect during these months
plays a significant role in variation of Ca/Cl ratio.
Magnesium: The data on level of magnesium in seawater
are given in Table 5 and Fig. 4. The surface values of mag-
nesium varied from 1.120 to 1.244 g/kg during pre monsoon,
1.186 to 1.172 g/kg during monsoon and 0.943 to 1.148
g/kg during post monsoon at Station-1; from 1.030 to 1.230
g/kg during pre monsoon, 1.208 to 1.217 g/kg during
monsoon and 0.900 to 1.170 g/kg during post monsoon at
Station-2; and from 0.872 to 1.118 g/kg during premonsoon,
1.238 to 1.302 g/kg during monsoon and 1.240 to 1.148 g/
kg during postmonsoon at Station-3. The bottom values var-
ied from 1.172 to 1.251 g/kg during premonsoon, 1.223 to
1.208 g/kg during monsoon and 0.968 to 1.163 g/kg during
postmonsoon at Station-1; from 1.001 to 0.913 g/kg during
premonsoon, 1.268 to 1.216 g/kg during monsoon and 1.230
to 1.223 g/kg during postmonsoon at Station-2; and from
0.799 to 1.251 g/kg during premonsoon, 1.349 to 1.221 g/
kg during monsoon and 0.894 to 1.194 g/kg during
postmonsoon at Station-3. In general, the values were rela-
tively higher in bottom waters compared to surface waters.
The high concentration of Mg in bottom waters is due to re-
mineralization taking place from the calcite skeletons con-
taining high Mg content. The adsorption of Mg through sus-
pended matter is one of the reasons for the decline in the
surface water concentration. However, lower values observed
during the monsoon and postmonsoon months were due to
 T. Padmavati et al.

Table 1: Salinity (ppt) of near-shore waters of Gopalpur. Table 4: Calcium to chlorinity ratio in near-shore waters of Gopalpur.

Seasons Station 1 Station 2 Station 3 Seasons Station 1 Station 2 Station 3

Months Surface Bottom Surface Bottom Surface Bottom Months Surface Bottom Surface Bottom Surface Bottom

Premonsoon Premonsoon
March 30.98 31.04 31.56 32.78 31.16 31.33 March 0.02369 0.02596 0.02044 0.02234 0.02209 0.02474
April 33.60 34.44 34.35 34.22 33.96 34.12 April 0.02312 0.02366 0.02046 0.02245 0.02262 0.02112
May 33.15 33.34 33.18 33.38 33.12 33.32 May 0.02185 0.02140 0.02008 0.02170 0.02200 0.02076
Monsoon Monsoon
June 32.92 33.44 33.10 33.22 32.52 33.13 June 0.02058 0.02106 0.02047 0.02095 0.01943 0.02046
July 31.48 32.16 31.81 32.44 31.55 31.94 July 0.02009 0.01967 0.02210 0.2116 0.01946 0.01924
August 29.78 30.18 30.03 30.68 28.56 29.14 August 0.02427 0.02311 0.02407 0.02368 0.02658 0.02359
September 29.04 29.48 33.10 30.16 28.96 29.16 September 0.02489 0.02546 0.02492 0.02367 0.02470 0.02455
Postmonsoon Postmonsoon
October 27.94 28.71 28.08 28.98 22.09 22.38 October 0.02723 0.02649 0.02593 0.02587 0.03224 0.03174
November 30.13 30.32 30.06 30.25 29.48 29.79 November 0.02230 0.02260 0.02417 0.02427 0.02330 0.02329
December 26.58 27.82 26.54 26.94 19.48 19.78 December 0.02434 0.02567 0.02432 0.02611 0.03822 0.02227
January 29.67 29.67 29.91 29.99 24.59 25.06 January 0.02016 0.02393 0.01964 0.01958 0.02301 0.02893
February 30.48 30.95 30.52 30.99 22.09 29.16 February 0.02199 0.02090 0.01972 0.02175 0.02352 0.02136

Table 5: Concentration of magnesium (g/kg) of near-shore waters of

Table 2: Chlorinity (ppt) of near-shore waters of Gopalpur.
Gopalpur.
Seasons Station 1 Station 2 Station 3
Seasons Station 1 Station 2 Station 3
Months Surface Bottom Surface Bottom Surface Bottom
Months Surface Bottom Surface Bottom Surface Bottom
Premonsoon
Premonsoon
March 17.13 17.18 17.47 17.59 17.25 17.34
March 1.148 1.172 1.170 1.223 1.148 1.194
April 18.61 19.06 09.01 18.93 18.79 18.89
April 1.238 1.309 1.262 1.303 1.294 1.304
May 18.36 18.46 18.37 18.48 18.32 18.45
May 1.244 1.251 1.230 1.230 1.240 1.251
Monsoon
Monsoon
June 18.23 18.52 18.32 18.38 18.01 18.33
June 1.186 1.223 1.208 1.216 1.302 1.349
July 17.42 17.79 17.60 17.96 17.47 17.67
July 1.325 1.423 1.224 1.299 1.225 1.226
August 16.47 16.70 16.62 16.98 15.80 16.11
August 1.267 1.306 1.106 1.137 1.232 1.240
September 16.07 16.30 16.45 16.69 16.03 16.13
September 1.172 1.208 1.217 1.268 1.238 1.221
Postmonsoon
Postmonsoon
October 15.46 15.89 15.54 16.04 12.22 12.38
October 0.943 0.968 0.900 0.913 0.872 0.894
November 16.68 16.77 16.63 16.73 16.31 16.49
November 1.011 1.074 1.108 1.100 1.178 1.186
December 14.71 15.39 14.68 14.90 10.78 10.94
December 0.918 0.986 0.942 0.902 0.673 0.705
January 16.42 16.42 16.55 16.60 13.60 13.86
January 0.982 1.016 1.012 1.018 0.780 0.799
February 16.87 17.13 16.89 17.15 15.86 16.15
February 1.120 1.163 1.030 1.001 1.118 1.138

Table 3: Concentration of calcium (g/kg) of near-shore waters of Gopalpur. Table 6: Magnesium to chlorinity ratio in near-shore waters of Gopalpur.

Seasons Station 1 Station 2 Station 3 Seasons Station 1 Station 2 Station 3

Months Surface Bottom Surface Bottom Surface Bottom Months Surface Bottom Surface Bottom Surface Bottom

Premonsoon Premonsoon
March 0.406 0.446 0.357 0.393 0.381 0.429 March 0.06704 0.06816 0.06697 0.06947 0.06655 0.06886
April 0.430 0.451 0.389 0.424 0.425 0.399 April 0.06656 0.06868 0.06644 0.06878 0.06887 0.06903
May 0.401 0.395 0.369 0.401 0.403 0.383 May 0.06785 0.06777 0.06696 0.06656 0.06769 0.06780
Monsoon Monsoon
June 0.375 0.390 0.375 0.385 0.350 0.375 June 0.06515 0.06604 0.06588 0.06616 0.07224 0.07359
July 0.350 0.350 0.389 0.380 0.340 0.340 July 0.07606 0.07993 0.06955 0.07227 0.06525 0.06938
August 0.400 0.386 0.400 0.402 0.420 0.380 August 0.07694 0.07838 0.06661 0.06702 0.07791 0.07697
September 0.400 0.415 0.410 0.395 0.396 0.396 September 0.07287 0.07417 0.07398 0.07597 0.07729 0.06200
Postmonsoon Postmonsoon
October 0.421 0.421 0.411 0.415 0.394 0.393 October 0.06093 0.06092 0.05792 0.05686 0.07144 0.07221
November 0.372 0.379 0.402 0.406 0.380 0.384 November 0.06001 0.06404 0.06663 0.06575 0.07216 0.07192
December 0.358 0.395 0.357 0.389 0.412 0.364 December 0.06241 0.06400 0.06431 0.06658 0.06252 0.06444
January 0.331 0.393 0.324 0.325 0.313 0.401 January 0.06017 0.06181 0.06115 0.06133 0.05735 0.05765
February 0.371 0.358 0.333 0.373 0.373 0.345 February 0.06639 0.06783 0.06098 0.05837 0.07055 0.07034

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 DISTRIBUTION OF CALCIUM AND MAGNESIUM IN COASTAL WATERS OF GOPALPUR
Table 7: Average values of chlorinity, calcium, magnesium and their ratios in Gopalpur, Andaman Sea and Visakhapatnam Harbour waters.
Chlorinity (ppt) Calcium (g/kg)
Observed in the 16.925 0.388± 0.006
present study
Reported * 17.8± 0.19 0.421± 0.00503
values ** 17.54 0.385 ± 0.007
*Values for Andaman Sea (Bay of Bengal); ** Values for Visakkhapatnam Harbour
the freshwater influx from rivers and creeks to the near shore
water.
Table 6 shows the monthly variation of magnesium to
chlorinity ratio at all the stations. For surface water the ratio
ranged from 0.06001 to 0.07694, 0.05792 to 0.07498 and
0.05735 to 0.07791, and for the bottom water from 0.06092
- 0.07993, 0.05686-0.07597 and 0.05765 to 0.07697 for Sta-
tion-1, Station-2 and Station-3 respectively. The bottom wa-
ter ratios were, in general lower than the surface water, which
may be attributed to the presence of comparative low salin-
ity in surface waters.
The variations in both calcium and magnesium concen-
trations are observed in the present investigation but statis-
tically not significant. Similarly, surface to bottom differ-
ence is also not significant and the minor differences no-
ticed in the present study may be due to some removal mecha-
nisms for surface waters. Bottom values are always higher
than surface. Extraction of calcium by living organisms from
surface water leading to lower calcium level in the surface
water and precipitation of calcium carbonate at bottom re-
gion could result in increased calcium concentration in bot-
tom water. Comparing the calcium and magnesium levels
during the three seasons, it is observed that the values are
always lower in the postmonsoon season than the
premonsoon with an intermediate value during monsoon.
These conditions in the postmonsoon may be related to the
biogeochemical processes prevailing in different locations.
The lowering of the levels of calcium in coastal waters dur-
ing the monsoon months is obviously due to dilution effect.
Freshwater run-off as a result of monsoonal rains could
account for the lowering of the calcium and magnesium con-
centration. Naik (1978) also described a similar situation in
near shore water of Goa. Higher concentrations during
premonsoon season appear to be largely due to recovery of
calcium for gradual attainment of normal oceanic conditions
and almost constant values of the concentration of calcium
at all the stations may be due to the attainment of stable equi-
librium conditions in the waters (Naik 1978). Sen Gupta et
al. (1978) reported that maximum recovery of calcium oc-
curs during premonsoon season which is more comparable
to oceanic ones.
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

Magnesium (g/kg) Ca/Cl Mg/Cl
1.163± 0.0008 0.02305± 0.0009 0.06799 ± 0.000172
1.268± 0.000176 0.02224± 0.000045 0.06655
1.203± 0.0011 0.02198± 0.000008 0.06856± 0.000008
The average Ca/Cl ratio is 0.02305 ± 0.00009 with an
average calcium concentration of 0.388 ± 0.0006 g/kg, the
average chlorinity being 16.93 ppt (Table 7). The Ca/Cl val-
ues are high when compared to those reported for Andaman
Sea (Bay of Bengal) and Visakhapatnam locality. Lower
Ca/Cl values are mainly due to comparatively lower salinity
(30.59 ppt) to that of reported values (Sen Gupta 1978, Sahu
1986). The average Mg/Cl ratio is 0.06799 ± 0.000172 with
an average magnesium concentration of 1.153 ± 0.0008
(Table 7). Average concentration of magnesium is lesser than
the reported values; whereas the ratio to chlorinity is higher
than Andaman Sea and lower than Visakhapatnam waters.
The scattered plots between chlorinity versus calcium
(Fig. 5) and chlorinity versus magnesium (Fig. 6) were drawn
and the least square method was adopted to draw the regres-
sion lines. The regression equation computed for calcium
with chlorinity is y = 0.0015 x + 0.3613, and magnesium
with chlorinity is y = 0.0041 x + 1.0023. It appears from the
calcium concentration graph (Fig. 5) that its concentration
ranges from 0.313 to 0.510, and magnesium concentration
from 0.673 to 1.423. The calcium versus chlorinity indicates
that the low ranges of calcium concentration were caused
due to the insignificant source of calcium, while the graph
(Fig. 6) on magnesium versus chlorinity indicates the wider
variations of magnesium concentration caused due to the bio-
chemical activity as well as addition of magnesium from
extraneous sources.
&21&/86,21
The major elements comprise of dissolved salts in seawater.
These are very few in numbers and the major cations and
anions play a significant role in seawater. These are con-
servative and bear a constant ratio with chlorinity. The con-
servative behaviour encountered was due to slight changes
in physical properties of seawater. The salinity varied from
19.78 ppt to 34.35 ppt. The calcium and magnesium varied
from 0.313 g/kg to 0.51 g/kg and 0.674 g/kg to 1.422 g/kg
respectively. The concentration of calcium and magnesium
is enriched with the influx of freshwater.
The scattered plots between chlorinity versus calcium and
magnesium were drawn on regression lines. The calcium
 T. Padmavati et al.

Salinity Station - 1

Station - 2

Station - 3

Fig. 2: Monthly distribution of salinity (ppt) of near shore water of Gopalpur Port at the three Stations from March 2007 to February 2008.

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 DISTRIBUTION OF CALCIUM AND MAGNESIUM IN COASTAL WATERS OF GOPALPUR 

Fig. 3: Monthly distribution of calcium concentration of near shore water of Gopalpur at the three Stations from March 2007 to February 2008.

Fig. 4 : Monthly distribution of magnesium concentration of nearshore water of Gopalpur Port at the three Stations from March 2007 to February
2008.

Fig. 5: The seasonal linear relationships of Ca with chlorinity. Fig. 6: The seasonal linear relationships of Mg with chlorinity.

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 T. Padmavati et al.
versus chlorinity indicated a low range of variability in con-
centration of calcium, and magnesium indicated a wider range
due to biochemical activity of organisms. Hence, the find-
ings of the present study indicate that there is monthly vari-
ation from time to time. The conservative behaviour of ions
(Ca++ and Mg++) does not show a deviation from the Marcets
principle of conservative behaviour of major ions in seawater
which reflects on the water quality of Gopalpur coast.
$&.12:/('*(0(17
The author T. Padmavati is thankful to the HOD of Marine
Sciences, Berhampur University for providing the labora-
tory facility to carry out the work. Thanks are also due to the
UGC, New Delhi for providing the financial assistance in
awarding JRF under the Rajiv Gandhi National Fellowship
(RGNF).
5()(5(1&(6
Culkin, F. 1965. The major constituents of seawater. In: Chemical Ocea-
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
nography, Vol. 1, Riley, J.P. Skirrow, G. (eds.), Academic Press, Lon-
don, 121-162.
Grashoff, K. 1976. Methods of Seawater Analysis. Verlag Chemie,
Weinhein, pp. 260.
Marcet, A. 1819. On the specific gravity and temperatures of seawaters in
different parts of the oceans and in particular seas with some account
of their saline contents. Phil. Trans., 161-208.
Naik, S. 1978. Calcium and magnesium concentration in the near shore
waters of Goa. Mahasager, Bull. Natl. Inst. Oceanogr., 11: 185-189.
Naronha, R.J., Caroline, M. and Sen Gupta, R. 1981. Calcium, magnesium
and chloride concentration in the Andaman Sea. Indian J. Mar. Sco.,
10: 234-237.
Sahu, S.D. 1986. Studies on Ca and Mg off Vishakapatnam Harbour, Bay
of Bengal. M.Phil Dissertation, Berhampur University, Berhampur,
Orissa.
Sen Gupta, R.S., Naik, S.S. and Singbal, S.Y.S. 1978. A study of fluoride,
calcium and magnesium in the northern Indian Ocean. Marine Chem-
istry, 6: 125-141.
Tripathy, S.K., Panigrahi, R.C., Gouda, R. and Panda, D. 1990. Distribu-
tion of calcium and magnesium in the Rushikulya estuary (Orissa),
East Coast of India. Indian J. Mar. Sci., 19: 212-214.
Wilson, T.R.S. 1975. Salinity and major elements of seawater. In: Chemi-
cal Oceanography, Vol. 1, Riley, J.P. and Skirrow, G. (eds.),
Academic Press, London, pp. 365-414.
 0CVWTG'PXKTQPOGPVCPF2QNNWVKQP6GEJPQNQI[ ,661 9RO 1R SS 
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B. Digamber Rao* G. Shamitha**, G. Renuka** and M. Ramesh Babu*
*Department of Botany, Kakatiya University, Warangal-506 009, A.P., India
**Department of Zoology, Kakatiya University, Warangal-506 009, A.P., India

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,1752'8&7,21
Cyanobacteria are a very old group of organisms and repre-
sent relics of the oldest photoautotrophic vegetation in the
world that occur in freshwater, marine and terrestrial habi-
tats (Mundt & Teuscher 1988). Cyanobacteria have drawn
much attention as prospective and rich sources of biologi-
cally active constituents and have been identified as one of
the most promising groups of organisms to be capable of
producing bioactive compounds (Schlegel et al. 1999). Re-
cent investigations on biologically active secondary
metabolites from cyanobacteria led to the identification of
wide range of compounds possessing antimicrobial, antivi-
ral, antineoplastic and toxic properties (Falch et al. 1995,
Moore 1996, Namikoshi & Rinehart 1996). The marine
cyanobacterial group is well documented for its bioactive
compounds possessing antimicrobial properties, while there
are only a few reports available with freshwater cyanobacteria.
In the present work, we report the C-phycocyanin, antifun-
gal activity and the toxicity evaluation of the culture filtrate
of four strains of Tolypothrix sp. and their impact on the
carbohydrate content and amylase activity of Eri silkworm,
Samia cynthia ricini.
0$7(5,$/6$1'0(7+2'6
Soil samples were collected from different agro-climatic re-
gions of paddy fields in North Telangana region of Andhra
Pradesh. Soil samples in laboratory were cultured in G 11
media with or without nitrogen source. The cyanobacteria
observed were transferred to the same medium. Unialgal
cultures were prepared using sub-culturing methods. Each
isolated cyanobacteria was cultured in a 500 mL flask con-
taining 150mL of BG 11 medium without shaking for 30
days. The incubation temperature was 28°C ± 2 and illumi-
nation at 3000 lux with a white continuous fluorescent light.
Preparation of supernatant and cell extracts: The cultures
were harvested after 30 days by centrifugation at 5000 rpm
for 15 minutes. The aqueous supernatant was collected and
the algal pellet was extracted with 15 mL of methanol, with
shaking for 20 minutes. The culture supernatants and sol-
vent extracts were dried under reduced pressure at 40°C and
were stored at -10°C for further studies.
The most commonly used method for the extraction of
phycobiliprotein C-phycocyanin was isolated following the
method of Boussiba & Richmond (1979) and estimation as
described by Bennett & Bogorad (1973). Ten mL of
cyanobacterial culture was homogenized and centrifuged at
5000 rpm for 5 minutes. The pellet was washed and sus-
pended in 2.0 mL of 0.05 M phosphate buffer (pH 6.8). The
aqueous phase containing cyanobacterial cells was subjected
to freezing and thawing. Then the content was centrifuged
at 5000 rpm for 5 minutes and the supernatant was collected
and stored in the refrigerator. Again the pellet was subjected
to freezing and thawing until a colourless supernatant was
obtained. The supernatant containing pigment was pooled
 B. Digamber Rao et al.
and final volume was observed. The pigment absorption was
measured at 615 and 652 µm in a Beckman DU-64
spectrometer against 0.05 M phosphate buffer as blank. The
concentration of C-phycocyanin was calculated using the
following formula. The phycobilin pigments C-phycocy-
anin content in the cyanobacterial cultures were expressed
as µg per mL of the cyanobacterial cells under investigation.
E -615-0.474(E 652)
C-Phycocyanin (PC ) = –––––––––––––––––
5.34
Where, E 615 and and E 652 are the absorbance at 615,
652 µm respectively.
Identification of cyanobacteria: Identification of the
cyanobacteria was done by using morphological variation
studies and taxonomical approaches mentioned in the pub-
lished literature of Desikachary (1959), Anand (1989) and
Santra (1993).
Antifungal bioassay: Antimicrobial activities of the C-phy-
cocyanin and extraction solvent of Tolypothrix sp. were de-
termined by the paper disk diffusion method. The following
fungi were used as test organisms: Candida albicans, Can-
dida guilliermondii, Aspergillus fumigatus and Aspergillus
niger. Filter paper discs (6mm) were saturated with 50 µL of
the test solution, dried under laminar air flow and placed on
the Saubouraud’s dextrose agar plate, which had been in-
oculated with a lawn of the test microorganisms. Plates were
incubated for fungi at 25°C for a period of 24-48 hours. Discs
treated with 50 µL methanol was used as negative control
and Gentamycin discs were used (10 µg) as positive con-
trols. The extracts and supernatants containing antifungal
components produced distinct and clear circular zones of
inhibition around the discs. The diameter of clear zones was
determined in millimetres and used as an indication of anti-
fungal activity. All tests were made in triplicate under ster-
ile conditions.
The following formula was used for comparison of the
antifungal produced activity of the sample with that of the
standard.
Inhibition zone of sample
Antimicrobial index = ––––––––––––––––––––––––– × 100
Inhibition zone of the standard
Silkworm toxicity assay: The present study also included
observations of the phycocyanin toxicity. Fifty number of
second instar larvae of Eri silkworm (Samia cynthia ricini)
were taken in a clean plastic tray of 20 cm diameter and 8 cm
height. A quantity of 50 g of clean tender and chopped leaves
of caster (Ricinus communis) was taken and one gramme of
the C-phycocyanin was dissolved in minimum quantity of
water and thoroughly mixed. The leaves were allowed to air
dry. After starvation for 5 hr, the larvae were allowed to feed
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
the treated leaves. Untreated leaves were also fed to another
set of larvae which served as a control. Observations were
made after 24 hr for the worm movement, death rate, feed-
ing rate and symptoms of ill health. The larval body weight
and gland weight of the silkworm were observed when the
larvae passed on to the late fifth instar from early fifth instar.
Estimation of carbohydrate was done (Mokrash 1954)
by collecting fifth instar larval haemolymph of both control
and treated worms and expressed in terms of mg/mL.
Haemolymph was collected from fifth instar silkworms by
cutting the last abdominal leg and collecting in precooled
tubes containing a few crystals of phenylthiourea. Digestive
fluid was collected from the 4th day of fifth larval stadium.
Larvae were starved for 4 h and digestive juice was collected
from mid-guts after dissection of larvae. Digestive fluid was
collected into precooled tubes. Five individual larvae were
used for sample collection. Since the sex was not discern-
ible during the larval stage, it is probable that the samples
were derived from both sexes. The digestive fluid was cen-
trifuged at 10,000 rpm for 10 min to remove undigested leaf
particles and stored at -20°C until use. Amylase activity was
measured with the standard procedure using soluble starch
as substrate (Bernfeld 1955, Baker 1991). Maltose was used
as a standard and the enzyme activity was expressed as mg
of maltose released/mL/min at 37°C. Samples were diluted
prior to assay to maintain linearity. Enzyme assays were car-
ried out for five individual larval samples calibrated against
controls for two trials and means of all the five values and
trials were taken as the final values (Abraham et al. 1992).
5(68/76
Certain cyanobacteria have drawn much attention as pro-
spective and rich sources of biologically active constituents
and have been identified as one of the most promising groups
of organisms capable of producing bioactive compounds
(Fish & Codd 1994, Schlegel et al. 1999). Although the po-
tential of cyanobacteria as biofertilisers is well known, at-
tention has recently been focused on the biotechnological
potentials of cyanobacteria for obtaining pharmacolog-ically
active secondary metabolites (Carmichael 1992). The ma-
rine cyanobacteria are well documented for their bioactive
compounds possessing antimicrobial properties, while there
are only few reports available with freshwater cyanobacteria.
Antifungal activity of culture filtrate and C-phycocyanin:
The results of solvent extracts and C-phycocyanin of the
selected strains of cyanobacteria Tolypothrix ceylonica, T.
fragilis, T. nodosa and T. tenuis were against the pathogenic
fungi (Candida albicans, C. guilliermondii, Aspergillus
niger and A. fumigatus) have been given in Table 1. In gen-
eral, the inhibition of culture extracts was more followed by
 ACTION OF C-PHYCOCYANIN AND CELL EXTRACTS OF TOLYPOTHRIX ON SILKWORM
the C-phycocyanin pigment. In the study of antifungal ac-
tivity T. fragilis culture extracts have shown higher inhibi-
tion (11.7 mm) followed by C-phycocyanin from T. tenuis
(9.4 mm) on the pathogenic fungi of Aspergillus fumigatus.
The culture extract from Tolypothrix tenuis (7.7 mm) and C-
phycocyanin from T. fragilis (6.6 mm) showed higher inhi-
bition of Aspergillus niger among the different treatments.
The culture extract of T. fragilis (11.6 mm) and C-phycocy-
anin of T. tenuis (9.4 mm) expressed higher inhibition on
the pathogen of Candida albicans under study. The Can-
dida guilliermondii appears to be more resistant by express-
ing the minimum inhibition zones for culture extracts (1.3-
4.8 mm) and C-phycocyanin (2.7-4.2 mm) when compared
with the control (Gentamycin).
In the present study the effects of C-phycocyanin pig-
ments of various strains of Tolypothrix ( T. fragilis, T. nodosa
and T. tenuis) on carbohydrate content in the haemolymph
of silkworms were found to be as 43.3 ± 1.4, 44.91 ± 1.2,
42.8 ± 2.8 and 49.5 ± 0.7 respectively and control was ob-
served as 40.8 ± 3.63 mg/mL. The percent increase in car-
bohydrate content in the haemolymph of silkworm treated
with the four strains of Tolypothrix was more (6.12, 10.04,
5.04 and 21.32) as compared to control under study (Table
2).
The digestive amylase activity values after treatment with
Tolypothrix ceylonica have expressed as 0.027 ± 0.001, fol-
lowed by Tolypothrix fragilis (0.033 ± 0), Tolypothrix
nodosa (0.026 ± 0.001) and Tolypothrix tenuis (0.03 ± 0.001
mg/mL) compare to control (0.026 ± 0.002 mg/mL/min).
The percentage increase of phycocyanin treated samples (T.
ceylonica, T. fragilis, T. tenuis) of digestive amylase activ-
ity of silkworm have shown 3.84%, 26.92%,0% and 15.38
respectively (Table 2) .
Silkworm toxicity test: Certain cyanobacteria have drawn
much attention as prospective and rich sources of biologi-
cally active constituents, and have been identified as one of
the most promising groups of organisms capable of produc-
ing bioactive compounds (Fish & Codd 1994, Schlegel et
al. 1999). Cyanobacteria are known to produce metabolites
with diverse biological activity such as antibacterial, anti-
fungal, antiviral, anticancer, antiplasmodial, algicide,
antiplatelet aggregation and immuno-suppressive activities
(Borowitzka 1995, Jaki et al. 2000, Kajiyama et al. 1998).
Patterson & Carmeli 1992, Gerwick et al. 1994, Luesch et
al. 2000, Papendorf et al. 1998, Papke et al. 1997, Rho et al.
1996, Koehn et al. 1992, Ghasemi et al. 2003). Few mem-
bers of Cyanobacteria are very toxic to human beings and
also to animals (Tyagi et al. 1999). The present study also
deals with observations of the toxicity cyanobacterial pig-
ment C-phycocyanin obtained from Tolypothrix sps. Toxic-
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

ity of phycocyanin pigments was tested in a sensitive insect
silkworm and the results were presented in Table 3. Phyco-
cyanin pigment from the four cyanobacterial cultures was
mixed individually with the mulberry leaves fed to the silk-
worm larvae once in a day caused no toxicity. The move-
ment of the worms was normal. No symptom of ill health
and abnormal secretion was detected. In all the treated worms
100 percent survival was observed. The body weight of the
silkworm and silk gland weight were increased when com-
pared to control worms ranging from 14.82-20.01% and
54.81-73.32% respectively.
',6&866,21
The cyanobacteria such as Fischerella ambigua (Flach et al.
1995), Fischerella musciola (Hagmann et al. 1996), Nostoc
commune (Jaki et al. 2000), Scytonema hofmanni (Pignatello
et al. 1983), Hapalosiphon fontinalis (Moore et al. 1987),
Anabaena sp. (Frankmolle et al. 1992), Nostoc spongiaeforme
(Hirata et al. 1996), Microcystis aeruginosa (Ishida et al.
1997), Phormidium sp. (Fish & Codd 1994) have been re-
ported as important cyanobacterial members to produce an-
timicrobial substances. These reports are in agreement with
the present study, since the extracts from Tolypothrix sps.
had similar effects on the pathogenic microbes used in the
present investigation.
Screening efforts aimed to identify antimicrobial agents
in cyanobacteria have revealed several promising com-
pounds. Some of these substances identified are Nostocyclyne
A (Plotono & Carmeli 2000), Nostofungicidine (Kajiyama
et al. 1998), Kawaguchipeptin B (Ishida et al. 1997), Nostocin
A (Hirata et al. 1996), Ambigol A and B (Falch et al. 1995),
Hapalindoles (Moore et al. 1987), and Scytophycins
(Ishibashi et al. 1986).
A few studies have been made to screen cyanobacteria
from paddy-fields for the production of antimicrobial sub-
stances. Possibly the synthesis of highly active toxin is a de-
fence option of cyanobacteria in these environmental condi-
tions against other organisms like bacteria, fungi, viruses and
eukaryotic microalgae (Mundt et al. 2001). Antimicrobial ac-
tivity of Tolypothrix sps. not yet been studied and the screen-
ing programme is among the first studies done for assessment
of its antibacterial and antifungal activity. The toxicity tests
are important for the algal products because, various members
of cyanobacteria produce toxins like cytotoxins, hepatoxins
and neurotoxins. In the present investigation C-phycocyanin
pigment from the four strains of Tolypothrix, T. ceylonica,
T. fragilis, T. nodosa, T. tenuis have not expressed toxicity
in the silkworm toxicity assay. The results on silkworm clearly
showed that the phycocyanin pigments are safe and also in-
crease body weight and silk gland weight under the study.
 B. Digamber Rao et al.

Table 1: Antifungal activity of cyanobacterial culture extract and C-phycocyanin.

Name of Inhibition annules (mm ± SEM)*

cyanobacteria Treatments Aspergillus Aspergillus Candida Candida
fumigatus niger albicans guilliermondii

Tolypothrix ceylonica Culture extract 11.4 ± 0.1 7.4 ± 0.1 8.1 ± 1.4 4.8 ± 0.2
phycocyanin 7.8 ± 0.4 5.2 ± 0.3 6.7 ± 0.5 4.2 ± 0.1
Tolypothrix fragilis Culture extract 11.7 ± 0.0 6.7 ± 0.2 11.6 ± 0.2 3.1 ± 0.0
phycocyanin 8.8 ± 0.3 6.6 ± 0.4 8.9 ± 0.4 2.7 ± 0.1
Tolypothrix nodosa Culture extract 6.7 ± 0.2 6.8 ± 0.5 11.4 ± 0.0 4.3 ± 0.2
phycocyanin 9.4 ± 0.0 4.9 ± 0.3 8.1 ± 0.2 3.0 ± 0.3
Tolypothrix tenuis Culture extract 8.8 ± 0.4 7.7 ± 0.5 8.0 ± 0.1 1.3 ± 0.5
phycocyanin 6.5 ± 0.3 5.5 ± 0.2 9.4 ± 0.4 2.8 ± 0.4
Gentamycin 13 ± 0.0 12 ± 0.0 14 ± 0.0 12 ± 0.0

*Data are presented as mean of three readings ± SEM.

Table 2: Effect of phycocyanin pigment on carbohydrate content in the haemolymph and digestive amylase activity.

Source of pigment Carbohydrate Percentage Digestive Percentage

content in increase (%) amylase activity increase (%)
haemolymph mg/mL/min
(mg/mL)

Tolypothrix ceylonica 43.3 ± 1.4 6.12 0.027 ± 0.001 3.84

Tolypothrix fragilis 44.91 ± 1.2 10.04 0.033 ± 0 26.92
Tolypothrix nodosa 42.8 ± 2.8 5.04 0.026 ± 0.001 -
Tolypotrix tenuis 49.5 ± 0.7 21.32 0.03 ± 0.001 15.38
Control 40.8 ± 3.63 - 0.026 ± 0.002 -

• The values are expressed in terms of standard error of the mean.

Table 3: Effect of phycocyanin pigment on the body weight and silk gland weight of Eri silkworm (Samia cynthia ricini).

Name of cyanobacteria Body weight of Percentage Silk gland Percentage

silkworm (mg) increase weight (mg) increase

Tolypothrix ceylonica 7447 ± 0.23* 20.01 1093 ± 0.13* 54.8

Tolypothrix fragilis 7634 ± 0.41* 15.345 1224 ± 0.21* 73.32
Tolypothrix nodosa 7125 ± 0.34* 14.829 1124 ± 0.22* 59.24
Tolypothrix tenuis 7339 ± 0.54* 18.27 1094 ± 0.26* 54.91
Control 6205 ± 0.48 - 706 ± 0.10 -

Values are mean ± SEM for ten larvae. *Statistically significant difference in comparison with the control group with p < 0.001

Trehalose is the major and metabolically active, non-
reducing disaccharide in the insect blood, which is synthe-
sized in the fat body, and utilized during spinning, flight
and starvation of insects (Saito 1960, Hori 1969). From the
present studies the increase in carbohydrate content in the
silkworms treated with phycocyanin extracted from various
Tolypothrix sps. suggests that carbohydrates are determinant
factor for the normal growth and development of the larva,
which ultimately determines the quality of silk produced.
The impact of biochemical parameters on yield revealed the
importance of amylase activity for the survival of silkworm,
Bombyx mori (Chatterjee et al. 1993). According to Hori
(1969), certain food compounds can stimulate or inhibit di-
gestive enzymes. Hirata & Yosuo (1974) found that
silkworm strains, which have more amylase activity showed
better cocoon weight, shell weight and shell percentage, and
rate of synthesis of amylase is dependent upon the rate of
feeding. Studies on quality of silk fibre through application
of rhizobacteria suggests that rhizobacteria could be an ef-
fective tool for enhanced biomass production in some plants,
which in turn has an impact on the growth of silkworms to
produce more silk fibre of good quality (Unni et al. 2008).
&21&/86,21
Many secondary metabolites are potent toxins, causing health
problems for animals and humans when the producer organ-
isms occur in masses in water bodies. The toxins produced
by cyanobacteria are grouped into three categories on the

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 ACTION OF C-PHYCOCYANIN AND CELL EXTRACTS OF TOLYPOTHRIX ON SILKWORM
basis of the bioassay methods used to screen them like
cytotoxins, hepatotoxins and neurotoxins. Cytotoxins are
studied with cultured cell lines, there are still no data on
cytotoxins from natural sources that are lethal to animals.
Recent investigation on biologically active secondary
metabolites from cyanobacteria led to the identification of
wide range compounds possessing antimicrobial, antiviral,
antineoplastic and toxic properties. The ability to produce
antimicrobial substances may be noticed not only as a de-
fensive instrument for the strains but also as a good source
of new bioactive compounds from pharmaceutical point-of-
view. The present study, aimed at the preliminary investiga-
tion of antifungal activity and toxicity evaluation studies
of Tolypothrix sp., indicated that this group of selected strains
displays a potential that warrants further work.
5()(5(1&(6
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digestive amylase from the tasar silkworm, Antheraea mylitta (Lepi-
doptera: Saturniidae). Comp. Biochem. Physiol., 110B (1): 201-209.
Namikoshi, M. and Rinehart, K.L. 1996. Bioactive compounds produced
by cyanobacteria. J. Ind. Microbiol., 17: 373-384.
Papendorf, O., Konig, G.M. and Wright, A.D. 1998. Hirridin B. and 2,4-
dimethoxy-6-heptadecylphenol, secondary metabolites from the
cyanobacterium Phormidium ectocarpi with antiplasmodial activity.
Phytochem., 49: 2383-2386.
Papke, U., Gross, E.M. and Francke, W. 1997. Isolation, identification and
determination of the absolute configuration of Fischerellin B. A new
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rahedron Lett., 38: 379-382.
 B. Digamber Rao et al.
Patterson, G.M.L. and Carmeli, S. 1992. Biological effects of tolytoxin (6-
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cyanobacteria. Arch. Microbiol., 157: 406-410.
Pignatello, J.J., Porwoll, J., Carlson, R.E., Xavier, A., Gleason, F.K. and
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Scytonema hofmanni. J. Org. Chem., 48: 4035-38.
Ploutno, A. and Carmeli, S. 2000. Nostocyclyne A, a novel antimicrobial
cyclophan from the cyanobacterium Nostoc sp. J. Nat. Prod., 63:
1524-1526.
Rho, M., Matsunaga, K., Yasuda, K. and Ohizumi, Y. 1996. A Novel
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308-309.
Santra, S.C. 1993. Biology of Rice Fields Blue-green Algae. Daya Pub-
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Schlegel, I., Doan, N.T., De Chazol. N., and Smith G.D.1999. Antibiotic
activity of new cyanobacterial isolates from Australia and Asia against
green and cyanobacteria. J. Appl. Phycol., 10: 471-479.
Soltani, Khavari-Nejad, Tabatabaei Yazdi, Shokravi and Feenandez-
Valiente 2005. Screening of soil cyanobacteria for antifungal and an-
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Tyagi, M.B., Thakur, J.K., Singh, D.P., Kumar, A, Prasuna, E.G. and Kumar,
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Biotechnol., 9: 9-21.
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*Department of Microbiology, Arts, Science & Commerce College, Kamrej Char Rasta, Surat, India
**C. G. Bhakta Institute of Biotechnology, Gopal Vidhyanagar, Tarsadi, India
***Department of Biosciences, Veer Narmad South Gujarat University, Surat, India

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,1752'8&7,21
Synthetic dyes are widely used in textile, rubber products,
enamel, plastic, cosmetic and many other industries (Guang
et al. 2005, Jo & Chia 2001). Textile processing industries
is a widespread sector in developing countries. Among vari-
ous processes in the textile industry, dyeing process uses
large volume of water for dyeing, fixing and washing proc-
ess (Husseiny 2008). Almost 106 tons of dyes are produced
annually around the world, of which azo dyes, characterized
by one or more azo groups (R1–N = N–R2) linking substi-
tuted aromatic structures, represent about 70% by weight
(Guang et al. 2005). Azo monochlorotriazine (MCT) fibre-
reactive dyes are commonly used for batch dyeing of
cellulosic fibres, but typically exhibit lower exhaustion and
fixation levels than other classes (Brent et al. 2006). Excess
dyestuff in process water is highly undesirable because of
environmental concerns (Andreas et al. 2004, Khalid et al.
2008), health hazards and aesthetical aspects. Colour is the
first contaminant in the wastewater, which should be recog-
nized and has to be removed before it is discharged in the
environment (Handaayani et al. 2007). Removal of colour
from dye containing wastewater is current issue of discus-
sion and regulation in many countries because of awareness
that water is valuable asset which should be protected (Casieri
et al. 2008). Nowadays physico-chemical methods used for
colour removal of the effluents are effective, but they show
disadvantages in terms of operational problems, high cost,
and sludge production (Kapdan et al. 2000, Lopez et al. 2004,
Kodam et al. 2005). Moreover, those methods use more en-
ergy and chemicals than biological processes. Because of
these disadvantages, in recent years a number of studies have
focused on microbial decolorization and degradation of azo
dyes (Handaayani et al. 2007). In the present study two diazo
monochlorotriazine dyes, namely reactive red 141 and reac-
tive red 120, were used. The objectives of the present study
were to isolate efficient dye decolourising bacteria and
optimizing various parameters for dye decolorization.
0$7(5,$/6$1'0(7+2'6
Dyes and chemicals: Reactive red 141 and reactive red 120
were procured from National Chemicals, Surat, Gujarat. The
various chemicals used in this study were of analytical grade
and procured from Hi-Media Pvt. Ltd., Mumbai.
Sample collection: Highly coloured effluent was collected
from a dyeing unit in the GIDC, Pandesara, Surat. The pH of
the effluent was 7.5. The effluent was collected in airtight
sterile plastic container and filtered through ordinary filter
paper to remove large suspended particles.
Enrichment of sample: The effluent sample was inoculated
with 50 mg/L of reactive red 141 and reactive red 120 and
 Naresh V. Butani et al.
incubated on rotary shaker (100 rpm) at 30°C. After 24 h
5% of inoculum was transferred to fresh effluent along with
reactive red 141 and reactive red 120. Three such transfers
were made.
Screening of dye decolourising microorganisms: After
third transfer, cell suspension from last enriched flask was
plated on the Bushnell Hass agar medium for screening of
dye decolourising microorganisms. Composition of BH agar
medium was (g/L), MgSO4 0.2, CaCl2 0.02, KH2PO4 1.0,
(NH4) NO3 1.0, FeCl3 0.05 supplemented with reactive red
141 (200 mg/L), pH 7.4. From that five bacterial colonies
were selected on the basis of formation of decolorization
zone surrounding the colonies.
Isolation and characterization of decolourisers: Out of
selected five decolourising colonies most promising bacte-
rial colony was selected on its capacity to produce largest
decolorization zone on BH agar plate containing dye. The
isolated bacteria were characterized by various morphologi-
cal and biochemical tests according to Bergey’s Manual of
Systematic Bacteriology.
Dye decolorization experiments: Dye decolorization by
isolated bacteria was tested in 250 mL Erlenmeyer flasks
with 100 mL BH medium containing 200 mg/L of each dye.
The sterilized medium was inoculated with the isolated bac-
terial culture of uniform cell density (optical density 1.0 at
550 nm). The medium-to-inoculum ratio (v/v) was 50:1.
Inoculated medium was incubated at 30°C on rotary shaker
at 100 rpm.
Spectrophotometric assessment of microbial dye decolori-
zation: After 24 h of incubation, 3 mL of medium was with-
drawn from each flask. Aliquots were centrifuged at 10,000
rpm in a centrifuge for 15 min to separate cell mass, clear
supernatant was used to measure the decolorization at ab-
sorbance maxima of dye (544 nm). Uninoculated mediums
were incubated as a control to check abiotic decolorization.
Experiment was performed in triplicate. Decolorization ef-
ficiency was expressed as percentage of decolorization and
was calculated using equation,
Decolorization (%) = AC - AT /AC × 100
Where AC is the absorbance of the control and AT is
average absorbance of the test samples.
To ensure that the change in pH of the dye solution had
no effect on the decolorization, the visible spectrum was re-
corded between pH 5.0 to 11.0, in which the pH did not show
any effect in spectrum.
Optimization of physico-chemical conditions for maximum
decolorization of dyes
Effect of different carbon sources on decolorization: Three
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different carbon sources, i.e. glucose, lactose and sucrose
were tested for decolorization at various concentrations, i.e.
0.2%, 0.5%, 1.0% (w/v). 2 mL of inoculum was inoculated
in 100 mL BH medium along with respective dyes and dif-
ferent concentrations of carbon source. All flasks were incu-
bated at 30°C on rotary shaker. Aliquot was removed from
each flask for the determination of decolourising activity at
different time intervals.
Effect of nitrogen sources on decolorization: Two nitro-
gen sources were tested for decolorization of dyes. The con-
centration of organic nitrogen (urea) and inorganic nitrogen
source (ammonium chloride) were 0.2%, 0.5%, 1.0% (w/v).
2 mL of inoculum was added to 100 mL of BH medium along
with respective dyes, 0.5% glucose and different concentra-
tions of nitrogen source. All flasks were incubated at 30°C
on rotary shaker. Aliquot was removed from each flask for
the determination of decolourising activity at different time
intervals.
Effect of pH and temperature on decolorization: Effect
of pH and temperature on decolorization was observed by
growing the isolate in the BH medium containing respec-
tive dyes having pH range from 5.0 to 11.0. In the same way
the effect of temperature was examined by growing cultures
at 25°C, 27°C, 29°C, 31°C, 33°C, 35°C, 37°C, 39°C, 41°C
by keeping the pH of the medium 7.4 for 7 days. Samples
were withdrawn at different time intervals and decolourising
activity was determined as described earlier.
5(68/76$1'',6&866,21
From the effluent sample collected from a dyeing unit, a
promising decolourising bacterial strain was isolated. This
strain formed a distinct clear zone on BH agar plate contain-
ing dye. To identify this bacterium, we investigated its mor-
phological and physiological properties using various bio-
chemical media. On the basis of results the isolate was iden-
tified as Enterobacter gergoviae (Table 1).
Microbial dye decolorization: The isolated strain was tested
for its capacity to remove the two dyes. Dyes were added to
BH medium at concentration of 200 mg/L as sole source of
carbon and nitrogen. The results indicate that the strain is
capable of decolourising the dyes up to 95% in 7 days. Rate
of decolorization is depicted in Fig. 1. The results show that
the isolated strain is effective in decolorization.
Optimization of culture condition: For the maximization
of decolorization of the dyes by the isolated strain, experi-
ments were conducted for the optimization of carbon source,
nitrogen source, pH and temperature.
Effect of different carbon sources: Three different carbon
sources, glucose, lactose and sucrose were tested for maxi-
mum decolorization by the isolated strain. Each carbon source
 BACTERIAL DECOLORIZATION OF DIAZO MONOCHLOROTRIAZINE DYES 

Fig.1: Microbial decolorization. Fig. 2(a): Effect of glucose. Fig. 2(b): Effect of lactose.

Fig. 2(c): Effect of sucrose. Fig. 2(d): Effect of urea. Fig. 2(e): Effect of ammonium chloride.

Fig. 2(g): Effect of pH.

Fig. 2(f): Effect of temperature.

was added at 0.2%, 0.5% and 1.0% in BH medium contain-
ing respective dye at 200 mg/L..
The strain is capable of decolourising the dyes in the pres-
ence of glucose at various concentrations. Complete decolori-
zation was observed when there was addition of 0.5% of
glucose (Fig. 2a).
There was no increase in the rate of decolorization when
lactose and sucrose were added (Figs. 2b & 2c). Maximum
percentage decolorization was observed when glucose was
used as carbon source at 0.5%.

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Naresh V. Butani et al.

Table 1: Physiological and biochemical characterization of the isolated bacteria.

Sr. No. Characteristics Result Sr. No. Utilization of Result

1 Gram Reaction Negative 13 L-Arabinose Positive

2 Cell Morphology Short Rods 14 Cellobiose Positive
3 Motility Positive 15 Dulcitol Negative
4 Pigmentation Negative 16 Glycerol Positive
5 Spore Formation Negative 17 Lactose Positive
6 Urea hydrolysis Test Positive 18 Maltose Positive
7 Indole Production Test Negative 19 Mannitol Positive
8 Methyl Red Test Negative 20 Raffinose Positive
9 Voges Proskauer Test Positive 21 Sucrose Positive
10 Gelatin Hydrolysis Test Negative 22 Trehalose Positive
11 Phenyl Alanine Deaminase Test Negative 23 Xylose Positive
12 Glucose Dehydrogenase Test Positive 24 D- Sorbitol Negative

Effect of different nitrogen sources on decolorization: Two
nitrogen sources, urea and ammonium chloride, were tested
for decolorization of dyes by the isolated strain, the results
of which are depicted in Figs. 2(d) & 2(e). BH medium
containing respective dyes was supplemented with 0.5%, and
0.2%, 0.5% and 1.0% of urea and ammonium chloride
respectively. Results suggest that the strain showed
maximum decolorization at concentration 0.2% of urea and
0.2% of ammonium chloride. The best decolorization was
observed at 0.2% of ammonium chloride.
Effect of temperature and pH on dye decolorization: The
effect of temperature and pH on the dye decolorization was
tested. It was found that a temperature of 31°C was optimum
for maximum decolorization (Fig. 2f). Decline in
decolorization activity at higher temperature more than 39°C
can be attributed to the loss of cell viability. Optimum pH
for maximum dye decolorization was 7.0 (Fig. 2g).
The present study has resulted in the isolation of a
bacterial strain which has capacity of decolourising diazo
mono- cholorotriazine dyes, reactive red 141 and reactive
red 120, thus showing the potential to be exploited as possible
candidate for bioremediation. Decolorization activity can be
enhanced by addition of glucose. The isolated strain can
decolourise dyes under wide range of pH and temperature,
which is the nature of effluent from dyeing industries.
5()(5(1&(6
Andreas, K., Angelica, E., Arthur, C. and George, M. G. 2004. Laccase-
catalysed decolorization of the synthetic azo dye diamond black PV
200 and of some structurally related derivatives. Biocatalyst and Bi-
otransformation, 22(5/6): 331-339.
Brent, S., Rebecca, B. and Harold, S.F. 2006. High affinity, high efficiency
fiber-reactive dyes. Color Technology, 122: 187-193.
Casieri, L., Varese, G.C., Anastai, A., Prigion, V. and Svobodova, K. 2008.
Decolorization and detoxification of reactive industrial dyes by im-
mobilized fungi Trametes pubescens and Pleurotus ostreatus. Folia
Microbiol., 53(1): 44-52.
Guang, F.L., Ji, T. Z., Jing, W., Zhi, Y.S. and Yuan, Y.Q. 2005. Bacterial
decolorization of azo dyes by Rhodopseudomonas palustris. World
Journal of Microbiology & Biotechnology, 22: 1069-1074.
Handaayani, W., Meitinirati, V. I. and Timotius, K.H. 2007. Decoloriza-
tion of acid red 27 and reactive red 2 by Enterococcus faecalis under a
batch system. World Journal of Microbial Biotechnology, 23:
1239-1244.
Husseiny, S.M. 2008. Biodegradation of the reactive and direct dyes using
Egyptian isolates. Journal of Applied Sciences Research, 4(6):
599-606.
Jo, S.C. and Chia, Y.L. 2001. Decolorization kinetics of a recombinant
Escherichia coli strain harboring azo-dye-decolorizing determinants
from Rhodococcus sp. Biotechnology Letters, 23: 631-636.
Kapdan, I.K., Kargi, F., MacMullan, G. and Marchant, R. 2000. Effects of
environmental conditions on biological decolorization of textile dye-
stuff by C. versicolor. Enzyme Microbial Technol., 26: 381-387.
Khalid, A., Arshad, M. and Crowley, D.E. 2008. Decolorization of azo
dyes by Shewanella sp. under saline condition. Applied Microbiology
and Biotechnology, 79: 1053-1059.
Kodam, K.M, Soojhawon, I., Lokhande, P.D. and Gawai, K.R. 2005. Mi-
crobial decolorization of reactive azo dyes under aerobic conditions.
World J. Microbiol Biotechnol., 21: 367-370.
Lopez, C., Valade, A.G., Combourieu, B., Mielgo, I., Bouchon, B. and Lema,
J.M. 2004. Mechanism of enzymatic degradation of the azo dye Or-
ange II determined by ex situ 1H nuclear magnetic resonance and
electrospray ionization-ion trap mass spectrometry. Anal. Biochem.,
335: 135-149.

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Department of Environmental Science, Pune University, Pune, Maharashtra, India
*Department of Biotechnology, Fergusson College, Pune, Maharashtra, India

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,1752'8&7,21 1. Oxygen depletion in the receiving boy of water.

The manufacture of paper from pulp is water-intensive in-
dustry. It involves conversion of raw materials into pulp by
digestion with various material and conversion of pulp into
paper. Raw material used in this industry is mainly cellu-
lose. The yield of pulp depends mainly on the raw material
as given in Table 1.
Two trillion gallons of wastewater are discharged
annually by the pulp and paper industry, much of which is
highly coloured. The brown coloured effluents discharged
by the pulp and paper industry result in poor water aesthetics
as well as cause harm and disturbance to the surrounding
aquatic environment. The receiving waters can potentially
experience an increase in temperature and a decrease in
photosynthesis as a direct result of the addition of these
brown coloured effluents. In production of 1 ton of paper
150 m3 of effluent is generated, which is heavily loaded with
organic compounds depending on nature of the raw material,
process and chemicals used, finished product, and extent of
water reused. The effluent generated at the pulping stage,
called black liquor, contains various compounds like
dissolved lignin and its degradation products, hemicelluloses,
resin acid, fatty acids, tannins and phenols (Ali &
Sreekrishnan 2001, Lara & Rodrguez-Malaver 2003,
Malaviya & Rathore 2007). These organic compounds are
also responsible for giving the effluent its characteristic dark
brown colour and toxicity. Thus, it is obligatory to treat the
effluent before disposal. Overall pollution effects of
discharging raw wastewater from a paper mail are:
2. Presence of undesirable colour, odour and taste in the
water.
3. Reduced photosynthesis.
4. Formation of blanket of suspended solids settling at the
bottom of the receiving body of water.
5. The death of fish.
6. Toxicity added to the aquatic life due to formation of
mercaptans, pentachlorophenol, sodium pentachloro-
phenate, etc.
These effluents are usually treated in aerated lagoons and
activated sludge by biological processes, which reduce the
chemical oxygen demand (COD) and the biochemical oxy-
gen demand (BOD), but are not efficient in reducing colour,
chlorophenols and high-molecular-weight chlorolignins
(Cammarota &Avaliacao 1991). Many studies have demon-
strated that the colour and the chlorinated organic compounds
of kraft effluents can be reduced by white-rot fungi, which
are considered promising for the treatment of these efflu-
ents (Cammarota &Avaliacao 1991). White rot fungi have
the capacity to metabolize both lignin and its derivatives and
therefore, have potentials in the lignin/phenolic wastewater
treatment (Eaton & Chang 1982). These organisms have abil-
ity to perform decolourisation as well as to reduce absorb-
able organic halides and chemical oxygen demand (COD)
(Livernoche et al. 1983). They produce extracellular
oxidative enzymes, like lignin peroxidase (LiP), manganese
peroxidase (MnP) and laccase. These enzymes are presented
as the most functional in the biodegradation of lignin.
 Fateneh Bajoul Kakani et al.
2%-(&7,9(6
• Development of an environmentally-friendly technology
and cost-effective way for treatment of pulp and paper
industrial effluent.
• Reduction of the environmental impact of wastewaters
from the pulp and paper industry by using microorgan-
isms (algae, bacteria, fungi) with special emphasis on
wastewater biodecolourisation, biodegradation and
bioremediation.
• Utilization of wastewaters from the pulp and paper in-
dustry to obtain valuable products such as enzymes
(xylanases), single cell protein (SCP) and/or high-value
fatty acids (gamma-linolenic acid).
• Reuse/recycling of treated effluent/feasibility studies.
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3$3(5,1'8675<())/8(17
High lignin content: The most important raw material for
the production of pulp and paper is wood. Main element
groups of wood are lignin, cellulose and hemicelluloses.
Mainly lignins are degraded, heavily modified, chlorinated
and finally dissolved in the effluent. Therefore, the effluent
from such a process is dark brown due to content of
chromophoric, polymeric lignin derivatives. The products
of lignin transformation, primarily lignin peroxidase (Lip),
are the main contributors of effluent colour in paper mill
wastewaters (Perez et al. 1998). However, lignin degrada-
tion is dependent on the presence of a readily metabolizable
co-substrate, such as glucose.
High absorbable organic halide (AOX) concentration:
The content of chlorinated substances in effluents are usu-
ally ensured as total organically bound chlorine (TOCl) or
high absorbable organic halide (Eriksson 1993). The mo-
lecular mass of chloroorganics has very serious effect when
toxicity, biodegradation and decolourisation of effluent are
examined (Martin et al. 1995).
Colour: Dyes and pigments are highly visible material. Thus,
even minor release into the environment may cause the ap-
pearance of colour, for example, in open waters, which at-
tracts the critical attention of public and local authorities.
Table 1: Yield of pulp from different sources.
Raw Material Yield (%)
Rags 70-80%
Esparto grassStraw 40-45%40-50%
Wood(using sulphite process) 40-50%
Waste paper, waste fibers, bagging 70-90%
Bamboo 40%
Jute 50%
Bagasse 50%
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In last few decades, the colour problems associated with
pulp and paper mill wastewater have got a pretty important
role in research (Dilek & Bese 2001). The colour causes an
aesthetic problem and contributes to the BOD (Bajpai &
Bajpai 1994). Pulp and paper wastewater treatment by the
various steps involving biological technologies are tended
towards removing BOD, colour is at best incompletely re-
duced (Perez et al. 1998). Moreover, the lignin derivative
colour remains firm and unchanged in spite of chlorinated
organic removal (Tarlan et al. 2002).
COD/BOD ratio: BOD represents only the organic matter
which is capable of being degraded/oxidized by microbes
whereas COD represents all the oxidizable matter, includ-
ing organic matter in any particular effluent (Marmagne &
Coste 1996). The low molecular weight fraction of the
chlorolignins is the main contributor to the effluent BOD
and acute toxicity. The high molecular weight chlorinated
compounds contribute little to BOD and acute toxicity due
to their inability to pass through cell membranes. They are
the major contributor to effluent colour, COD and chronic
toxicity (Eriksson et al. 1985).
Potential toxicity problems: The low molecular weight
phenolics and their methylated counterparts, which are more
lipophilic, cause toxicity and are bioaccumulable in fish
(Kolar et al. 1985). However, Archibald et al. (1990) have
reported that toxic levels of low-molecular weight chlorin-
ated compounds do not accumulate during the natural deg-
radation of chlorolignins. They found that the effluents in-
deed appear to stimulate the growth of algae and primary
consumers, probably because of the nutrients they contain.
About 75% of the dissolved organic material, 60% of the
COD load, 40 to 50% of the organically bound chlorines,
and 80% of the colour-imparting substances of bleach plant
effluents are reportedly contributed by extraction stage ef-
fluents. The organic compounds in these effluents, mainly
the chlorinated phenols, are toxic to aquatic organisms and
resistant to microbial degradation.
Characteristics of raw wastewater: The wastewater is usu-
ally alkaline in nature and has high suspended solids, high
total solids, high COD and relatively low BOD. The approxi-
mate analysis of wastewater from typical pulp and paper mill
is given in Table 2.
Treatment of pulp and paper industrial effluent by bac-
teria and ligninolytic fungi: Over the past decade, many
fungal strains have been studied for their abilities to degrade
a wide variety of structurally diverse pollutants. White-rot
fungi produce a large variety of extracellular enzymes like
laccase, lignin peroxidase, phenol oxidase, Mn dependent
peroxidase and Mn-independent peroxidase, which decom-
pose the highly stable natural compounds such as lignin,
 ROLE OF MICROORGANISM IN TREATMENT OF PAPER AND PULP INDUSTRY EFFLUENT
hemicellulose, cellulose, etc. (Moreira et al. 2003). For that
purpose, white-rot fungi show as a valuable alternative be-
cause they are capable of oxidising compounds of complex
structure such as lignin (Kirk & Farrell 1987). This feature
is based on the extracellular secretion of highly oxidative
oxidases and peroxidases; the most outstanding enzymes
being lignin peroxidase (LiP) and manganese peroxidase
(MnP) (Tien & Kirk 1988). The treatment of pulp and paper
mill effluent has been studied using a number of fungi from
quite some time. the enzyme system of the white rot fungi
includes a group of nonspecific extracellular enzymes, which
catalyse not only degradation of lignin and chlorolignins but
also oxidation of several constant aromatic and halogenated
compounds like lindane, DDT, PCP, benzopyrene, creosote,
coal tars and heavy fuels, etc. (Eaton et al. 1982).
There are several investigations of potential ability of
fungi for treatment of pulp and paper wastewater effluent.
Recent studies using ligninolytic enzymes from filamentous
fungi as Aspergillus fumigatus and A. flavus showed a re-
moval from the paper effluent colour (Sahoo & Gupta 2005).
In this case, A. fumigates was able to decolorize 55.5% un-
der stationary condition and 89.3% under shaking condi-
tions, while A. flavus removed 53.5 and 84.0%, respectively,
in the same conditions. The difference in the colour reduc-
tion was related to differences in aeration between station-
ary and shaking flasks conditions. Basidiomycetes were also
applied in the paper effluent aiming decoloration, as
Pleurotus sajor-caju, P. platypus and P. citrinopileatus
(Ragunathan & Swaminathan 2004). After 6 days, P. sajor-
caju decolorized 66.7% of the effluent. These studies show
a decolorization between 55% and around 90%. However,
in order to obtain such efficiencies, it is necessary 3 to 6
days of treatment, which cannot be applied industrially.
Aspergillus foetidus was studied for its ability to remove
colour, COD and lignin from bagasse-based pulp and paper
mill wastewater (Sumathi & Phatak 1999). Nearly 90-95%
of the total colour was removed, and the simultaneous re-
duction in colour and lignin level indicated a strong correla-
tion between the decolorization and lignolytic processes. The
degradation abilities of Oxysporus sp., Phanerochaete
chrysosporium and Schizophyllum commune were evaluated
for the removal of lignin from olive pomace (Haddadin et
al. 2002). Among the three fungi, Oxysporus sp. was the most
Table 2: Characteristic of raw wastewater.
Raw Material Yield (%)
pH value 8.0-9.0
Total solids, mg/L 1500-2500
Suspended solids, mg/L 600-1500
COD, mg/L 300-2500
BOD5, mg/L 150-1000
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

effective for lignin degradation with efficiency of 70%,
whereas P. chrysosporium achieved lignin removal of 60%.
The effluent generated at the pulping stage (black liquor)
contains various compounds like dissolved lignin and its deg-
radation products, hemicelluloses, resin acid, fatty acids,
tannins and phenols (Ali & Sreekrishnan 2001, Lara &
Rodrguez-Malaver 2003, Malaviya & Rathore 2007).
Several researchers have examined biological treatment
of black liquor by using various pure bacterial strains. The
degradation efficiency of lignin in black liquor was 70-80%,
and COD removal efficiencies of 70-80% were achieved with
Pseudomonas putida (Sahoo & Gupta 2005) and
Acinetobacter alcoaceticus (Jain et al. 1997). In a batch test
on the treatment of black liquor from a Kraft pulp and paper
mill by Aeromonas formicans, around 71% of COD and 78%
of lignin were removed (Waraporn et al. 2006). Previous
work has focused mainly on the decolorization of black liq-
uor and the degradation of lignin by suspended- growth of
white rot fungi (Garcia et al. 1987). Kerem et al. 1992).
Among the biological methods tried so far, in this review
we comprise some of current investigation (Table 3) using
wood-degrading white rot fungi, which have the potential
to successfully treat pulp and paper effluents. In case of pulp
and paper industries, Phanerochaete flavidoalba (Perez et
al. 1998), Coriolus versicolor, Aspergillus niger
(Khanongnuch et al. 2004) and Schizophyllum commune
(Kannan & Oblisami 1990) have shown to be able to re-
move the residual recalcitrant colour efficiently from paper
and pulp mill effluents. Some white rot strains such as
Ceriopsis subvermispora could decolourize Kraft-bleaching
effluent at 90% and also resulted in reduction of COD up to
45%. However, higher percentages of COD, BOD and total
solids reduction in pulp mill wastewater have been reported
using a reduction-biological technique (Ghoreishi &
Haghighi 2007).
Santos et al. (2002) used the ligninolytic fungus
Pleurotus ostreatoroseus to treat the first alkali extraction
stage (E1) effluent of a Kraft bleach plant. The best results
were obtained with 20% fungus and shaking of the effluent.
During the last three days of the experiment, the average
percent removal of colour, total phenols and lignin/
chlorolignin was 84.4±6.1, 82.1±5.7 and 72.4±8.9, respec-
tively. The results indicate the potential of Pleurotus
ostreatoroseus for use in the treatment of E1 effluent (Santos
et al. 2002). In previous works some other authors used di-
luted E1 effluent without an extra carbon source and the fun-
gus Lentinus edodes, and obtained 73% colour removal af-
ter five days of treatment (Esposito et al. 1991). Livernoche
obtained 60% decolorization using fungus Coriolus
versicolor in liquid culture, and 80% with the fungus immo-
bilized on calcium. They used E1 effluent enriched with gluof
 Fateneh Bajoul Kakani et al.

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
ROLE OF MICROORGANISM IN TREATMENT OF PAPER AND PULP INDUSTRY EFFLUENT 

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Fateneh Bajoul Kakani et al.
Taguchi approach indicated optimum conditions: tempera-
ture (30-35 C°); shaking condition (125 rpm); dextrose (1.0%
w/v); tryptone (0.1% w/v); inoculum size (7.5%
w/v); pH (5) and duration (24 h). Overall evaluation crite-
rion (OEC) value before optimization was 32.3. There was
38% improvement in the process with final OEC value,
44.6±2.02 at optimum conditions. The colour content of the
effluent reduced by 50-53% and lignin content 35-40% af-
ter treatment at optimum conditions. Variation in pH from 5
to 6 has most significant effect on decolourisation (72%)
while variation in temperature from 30°C to 35°C has no
effect on the process. Treated effluent was further evaluated
for toxicity by alkaline single cell (comet) gel electrophore-
sis (SCGE) assay using Saccharomyces cerevisiae MTCC
36 as model organism, indicated 45% reduction. The results
showed significant reduction in colour, lignin and toxicity
of the effluent and this process can be scaled up to industrial
level. However, very few studies have optimized the proc-
ess. Studies related to the interaction of various factors among
them are lacking. For Cryptococcus sp., there was no sig-
nificant effect of temperature variation (30-35°C) on efflu-
ent treatment. Hence, temperature was dropped from the
analysis. The most suitable shaking condition was 125 rpm.
One possible reason may be very rapid movement leads to
the shear and tear of fungal mycelium (Mandels & Stenberg
1976, Broche-Due et al. 1994).
Sehanat et al. (2009) collected eight isolates, which could
be cultured to test for thermotolerance and to screen for the
presence of ligninolytic enzymes using Phanerochaete
chrysosporium as a reference. Of the eight isolates, only three
species Daedaleopsis sp., Schizophyllum commune PT, and
S. commune SL were able to grow above 40°C. All the three
species exhibited 2, 20-azino-bis(3-ethylbenzothiazoline-6-
sulphonic acid) (ABTS)-oxidizing activity on ABTS agar
plates. Only P. chrysosporium could oxidize manganese on
MnCl2 agar plates. Both P. chrysosporium and Daedaleopsis
sp. were able to decolourize wastewater on wastewater agar
plates. All fungal cell suspensions tested decolourized
wastewater No. 1 (pH 8.07, COD 4347 mg/L) from the
pulping process, and wastewater No. 2 (pH 6.94, COD 4000
mg/L) from the pulping process combined with that from
the paper recycling process. Daedaleopsis sp. and P.
chrysosporium exhibited the highest ability to decolourize
wastewater No. 1 (52%) and No. 2 (86%), respectively.
Laccase activities were detected in the decolourized effluents
and all fungi tested reduced the COD by 59-71% (No. 1)
and 66-83% (No. 2). The use of bacterial enzymes for effluent
colour removal in the paper industry was reported previously
(Joyce et al. 1984, Chairattanamanokorn et al. 2006, Nimchua
et al. 2008). Bacterial aerobic and anaerobic treatment
systems can reduce the BOD and are able to remove the dark
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
colour in the effluents as verified previously
(Chairattanamanokorn et al. 2006, Nimchua et al. 2008). In
a current study, Patricia Lopes de Oliveira et al. (2009)
applied Bacillus pumilus and Paenibacillus sp. on the paper
mill effluent to investigate the colour removal. Inocula were
individually applied in effluent at pH 7.0, 9.0 and 11.0. The
real colour and COD removal after 48h at pH 9.0 were
41.87% and 22.08% for B. pumilus treatment and 42.30%
and 22.89% for Paenibacillus sp., respectively. Gel
permeation chromatography was used to verify the molar
masses of compounds in the non-treated and treated effluents
showing a decrease in compounds responsible for the paper
mill effluent colour.
&21&/86,21
In general, the high volumes and complexity of effluent pro-
duced by the pulp and paper industry present formidable
problems for the treatment of these wastewaters. Biological
treatment methods that utilize ligninolytic microorganisms,
especially white-rot fungi, which are able to degrade lignin,
cellulose, and hemicellulose concurrently at approximately
equal rates. These fungi secrete three extra cellular enzymes,
which are critical for wood degradation, but the requirement
for a co-substrate and the need for aeration would seriously
hamper their utilization from an economic perspective.
Cultivation conditions have an important impact on the
efficiency of white-rot fungi during biological treatment of
bleach plant effluent. Greater biodegradation efficiencies are
required in order to develop a practical biotreatment proc-
ess. Any way, the possibility of wastewater bioremediation
prior to wastewater discharge has been demonstrated. The
use of continuous biosystems such as trickling filters, rotat-
ing biological contactors and activated sludge systems have
been shown to reduce the environmental impact of these
wastewaters. The most efficient treatment system was proved
to be the rotating biological contactor where colour, bacte-
rial growth inhibition levels, adsorbable organic halogen and
chemical oxygen demand were decreased to a significant
extent. Definite differences in the decolouring mechanisms
between the white-rot fungi (adsorption + biodegradation)
and the mucoralean fungi (adsorption) have been observed.
Because of the complexity of treating paper mill efflu-
ent, combinations of physical, chemical and biological treat-
ment strategies might lead to a synergistic beneficial out-
come that would facilitate the development of economic and
efficient treatment procedures.
5()(5(1&(6
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Department of Microbiology, Institute of Science, 15 Madame Cama Road, Mumbai 400 032, Maharashtra, India

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,1752'8&7,21 not all structurally related molecules. Also, individual mi-

The bane of industrial progress has been the generation and
release into the environment of huge amounts of toxic com-
pounds and xenobiotics like polyaromatic hydrocarbons
(PAHs) which have caused widespread contamination of land
and water. PAHs besides being persistent are toxic, possess
low volatility and aqueous solubility, hydrophobic in na-
ture, and have high solid water distribution ratios and high
affinity for the sediments which stand against their ready
microbial utilization (Johnsen et al. 2005). Because of the
great number of petroleum contaminated sites requiring
cleanup and the high cost involved with conventional ap-
proaches, there is an overriding need for research aiming at
establishing cost-effective and environmentally acceptable
mitigation technologies, primarily for the PAHs ranging in size
from naphthalene (two rings, C10H8) to coronene (seven rings,
C24H12) (Marta et al. 2006).
One such remedial technology is bioremediation that uses
microorganisms to detoxify environmental pollutants and trans-
form them into simpler, less toxic compounds. Microorgan-
isms, especially bacteria, play an important role in the bio-
degradation of these hydrocarbons. It is now realized that
microbial metabolism provides a safer, more efficient, and
less expensive alternative to physico-chemical methods for
pollution abatement. The high solubility of naphthalene com-
pared to other PAHs and the fact that the naphthalene
degradative genes are plasmid encoded has facilitated re-
search on naphthalene degradation. It is often used as a model
compound for in situ biodegradation studies of PAHs
(Grimm & Harwood 1999). It has been shown that the
acclimation of a microbial community to one substrate fre-
quently results in the simultaneous acclimation to some, but
crobial species have the ability to act on several structurally
similar substrates, and therefore, more easily act on their ana-
logues after the first addition. These model organisms may be
very useful for the study of complex aromatic hydrocarbon deg-
radation and for bioremediation purposes (Nnamchi et al. 2006).
Toluene and naphthalene degrading bacteria, obtained from
contaminated lake sediment, were used to degrade both
monoaromatics and polycyclic aromatic hydrocarbons
(PAHs) (Horng et al. 2009). Tikilili & Chirwa (2009) suc-
cessfully used naphthalene degrading bacterial species such as
Pseudomonas aeruginosa, Microbacterium esteraromaticum
and Alcaligenes sp. isolated from contaminated soil from a
landfill site (Chloorkop, Johannnesburg, South Africa) in de-
grading naphthalene found in radioactive wastewater collected
from radioisotope processing facility, South Africa. Thus, naph-
thalene degrading isolates could be very helpful in
bioremediating PAH-contaminated sites.
Hence, the aim of the present study was to optimize the
conditions for the degradation of naphthalene by the naph-
thalene degrading organisms, isolated from petroleum con-
taminated soils and to evaluate their potential for the
bioremediation processes.
0$7(5,$/6$1'0(7+2'6
All the media chemicals (AR grade) and reagents were pur-
chased from Hi Media Laboratories, Mumbai, India.
Organisms: The present study was detailed on Burkholderia
glathei, Alcaligenes denitrificans and Pseudomonas putida
isolated by enrichment culture technique from petroleum
contaminated soils in Mumbai, India. The isolates were
preserved on nutrient agar slants at 4°C.
 Varsha K. Vaidya and Sharmila Kadam
Optimization of conditions for naphthalene degradation:
Conditions for the degradation of 1% naphthalene were
optimized with respect to various environmental factors such
as pH, temperature, salinity, and addition of nutrients such
as nitrogen and phosphorus. The cultures for optimiza-
tion were prepared as follows: 18 h old isolates grown
on LB medium (g/L, Tryptone 10, Yeast extract 5, NaCl
5, D/W 1000 mL, pH 7.4) were transferred to Mineral
Salt Medium (MSM) agar plate (g/L, NH 4Cl 2, KH2PO 4
0.4, CaCl2 2, FeSO4. 7H2O 0.2, D/W 1000 mL, pH 7.4)
with naphthalene supplied in the vapour form from the
crystals placed in the lid of the plate. The plate was incu-
bated at room temperature (28 ± 2°C) for 48 h. Inoculum
was prepared by transferring one loopful of the growth from
the MSM agar plate to MSM liquid medium containing 1%
naphthalene dissolved in dimethyl formamide (DMF). Cells
were allowed to grow overnight to an optical density of
1.0 at A600. Ten percent of the 18 h old inoculum was
added to 50 mL of sterile MSM medium containing 1%
naphthalene as the sole source of carbon. The control flask
was prepared by adding 1 mL of the solvent DMF alone
(without naphthalene) and equivalent saline instead of
the culture (Malakul et al. 1998). Flasks were incubated
on shaker (120 rpm) at room temperature (28 ± 2°C) for
48 h. At the end of the incubation period, medium was cen-
trifuged at 3000 rpm for 10 min. Supernatant was taken and
residual naphthalene was estimated by measuring optical
density using UV spectrophotometer at 276 nm. A standard
curve of naphthalene was also plotted in the range of 100-
1000 µg/mL. The residual naphthalene after 48 h of incuba-
tion for all the parameters used for optimization was esti-
mated by UV spectrophotometer (Shimadzu UV1800 UV/
VIS) at 276 nm.
Naphthalene degradation was studied over a pH range
6.0-9.0. The pH supporting the maximum degradation of
naphthalene was used in further work. The isolates were
further tested for their ability to grow and degrade naph-
thalene at 16°C at pH optimized in the earlier step. The
impact of salinity on degradation was evaluated using
various concentrations of NaCl (0.5, 1.0, 1.5, 2.0, 2.5 and
3.0%) under the optimized conditions. The effect of dif-
ferent sources of nitrogen and phosphorus on the degra-
dation of naphthalene was studied as follows: Three sets
of six flasks each containing varying concentrations of
ammonium chloride, ammonium nitrate and urea (5-25 and
50 mg nitrogen/100 mg naphthalene) in 50 mL of sterile
MSM medium were used to study the effective nitrogen
source for achieving the highest degradation of naphthalene.
Two sets of six flasks each containing varying concentra-
tions of K2HPO4 and Na2HPO4 (0.07-0.12 mg phosphorus/
100 mg of naphthalene) in 50 mL of sterile MSM medium
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
were used to evaluate the effect of phosphorus on degrada-
tion of naphthalene. The effect of Fenton’s reagent (5 mL
2.8 M H2O2; 5 mL 0.1 M FeSO4) on degradation was evalu-
ated by adding 5 mL of Fenton’s reagent to 50 mL of MSM
broth inoculated with ten percent of inoculums under the
optimized conditions.
Application of the naphthalene degrading organisms in
soil: Degradation of the naphthalene in soil microcosm was
determined as follows: Thirty g of moist soil was placed in
500 mL flask containing 100 mL of MSM broth with 1%
naphthalene. The soil-naphthalene complex was mixed thor-
oughly and allowed to equilibrate for 72 h. 10 mL Fenton’s
reagent was added to naphthalene contaminated soil, the
contents were gently mixed and inoculated with individual
cultures and incubated at 28 ± 2°C. The residual concentra-
tion of naphthalene in soil was determined over a period of
seven days. An aliquot of the soil slurry with an equal vol-
ume of hexane was mixed in a separator funnel, mixed for
two minutes followed by extraction in cold conditions. The
resulting organic phase was used for the spectrophotometric
analysis of naphthalene (Marta et al. 2006).
Use of immobilized cells in degradation of naphthalene:
The 18 h old cells of the isolates grown in MSM medium
containing 1% naphthalene were harvested by centrifuga-
tion and washed twice in sterile distilled water. Alginate
beads containing immobilized cells were prepared by dis-
solving 4 g of alginate in 100 mL of distilled water over-
night and sterilized by autoclaving (121°C, 15 min). Ten
percent (optical density of 0.1 at A 540) of inoculums was
resuspended in the sodium alginate solution, stirring to
complete homogenization. The mixture was extruded
drop-wise through a hypodermic needle into a cold, ster-
ile solution of CaCl2 (0.2 M). The beads were left to harden
in the same solution at room temperature with gentle stir-
ring for 1 h. Finally, the beads were washed several times
with 0.9% NaCl to remove excess calcium ions and untrapped
cells. Sterile beads (without microorganisms) were used to
monitor the abiotic loss of naphthalene (Feijoo-Siota et al.
2008). The concentration of residual naphthalene in the cul-
ture medium was determined over a period of seven days by
measuring the optical density at a wavelength of 276 nm
after cold extraction with hexane (Marta et al. 2006).
Effect of storage on the viability of immobilized cells: Cell
viability of immobilized cells stored at 4°C was analysed
after 15 days by suspending the beads in10 mL of a sterile
1% solution of sodium citrate. The suspension was vortexed
to achieve a complete dissolution of the alginate. Serial di-
lutions were spread on LB agar. The colonies were counted
on LB agar plates after incubation for 48 h at 28 ± 2°C
(Feijoo-Siota et al. 2008).
 OPTIMIZATION OF CONDITIONS FOR NAPHTHALENE BIODEGRADATION
5(68/76$1'',6&866,21
Microbial communities present in the contaminated soils can
metabolize PAHs at greater rates than soil microbial com-
munities found in uncontaminated soils. Bacteria exposed
to pollutants can adapt to them by mutating or acquiring
degradative genes. These bacteria can proliferate in the en-
vironment as a result of the selection pressures created by
the pollutants. The positive outcome of selection pressure is
that harmful compounds may eventually be broken down
completely through biodegradation (Alquati et al. 2005, Van
der Meer 2006).
As suggested by Feijoo-Siota et al. (2008), a slightly al-
kaline pH seems to be quite favourable for hydrocarbon deg-
radation. In the present investigation, the range of pH for
degradation of naphthalene was varied between 6.0 and 9.0.
The extent of degradation for all the three bacterial isolates
increased over a pH range of 6.0-8.0 with pH 8.0 showing
the highest degradation for all the isolates. There was a dras-
tic reduction in the degradation ranging between 60-68 % at
pH 9.0. The degradation was the most adversely affected at
the acidic pH of 6.0 for all the isolates, though the effect was
pronounced in Burkholderia glathei, which showed a 88.05
% reduction in degradation compared to the degradation
obtained at optimal pH (8.0) (Fig. 1). Dibble & Bartha (1979)
observed maximum degradation rate at pH 7.8 in oil sludge
samples, beyond which rate of degradation decreased.
Rhodococcus rhodochrous KUCC 8801 and KUCC 8802
isolates showed their maximum growth at pH 8.0 in a crude
oil containing medium kept at 40°C. Thavasi et al. (2007)
also showed similar results, where maximum crude oil deg-
radation occurred at pH 8.0, beyond that level a decreasing
trend was observed. Pathak et al. (2008) showed that naph-
thalene degradation by Pseudomonas sp. HOB1 was aug-
mented in the pH range of 7.5-8.5.
Temperature plays a very important role in the biodeg-
radation of petroleum hydrocarbons, firstly by its direct ef-
fect on the chemistry of the pollutants and secondly by its
effect on the physiology and diversity of the microbial mi-
lieu. A temperature increase decreases viscosity and increases
diffusion rates of organic compounds. Therefore, higher re-
action rates due to smaller boundary layers are expected at
elevated temperatures. Increasing temperatures facilitate the
solubility of PAHs and hence their bioavailability. Although
microbial activity is generally reduced at low temperatures,
many of the components in crude oil can actually be degraded
in these conditions (Atlas 1981). Cold-tolerant isolates of
Pseudomonas sp. from oil polluted Antarctic soils have been
reported to utilize naphthalene, phenanthrene and fluorene,
as well as BTEX, as the sole carbon and energy source
(Aislabie et al. 2006). Jeon et al. (2004) reported
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

Polaromonas naphthalenivorans sp. strain CJ2T, capable of
growth on naphthalene at 4°C and 25°C (optimum, 20°C).
In the present investigation, the biodegradation of naphtha-
lene at 28 ± 2°C was compared to that at 16 ± 1°C in order to
evaluate the potential of the isolates to be used as
bioremediating agents in temperate conditions. The results
indicated that at 16 ± 1°C the rate of degradation decreased
for all the isolates viz. for Burkholderia glathei from 32.55%
to 10.33% (a decrease of 68.27 %), for Alcaligenes
denitrifacans from 36.25% to 12.93% (a decrease of 64.33%)
and for Pseudomonas putida from 38.96% to 16.87% (a de-
crease of 56.69%). However, degradation did not cease com-
pletely indicating the ability of the isolates to grow and de-
grade naphthalene even under lower temperatures.
Salinity is a major factor which significantly affects bio-
logical activity in the marine environment. There are few
published studies which deal with effects of salinity on the
microbial degradation of hydrocarbons. In the study by
Thavasi et al. (2007) Pseudomonas aeruginosa showed maxi-
mum growth and biodegradation activity at 3.5% salinity.
In the present study, the MSM medium used in the earlier
steps did not contain NaCl. When the medium was supple-
mented with NaCl in concentrations ranging from 0.5 % to
3% with an interval of 0.5%, it was observed that addition
of 0.5% salt did not alter the rate of degradation for any of
the isolate (Fig. 2). Except for a slight increase in degrada-
tion, shown by Burkholderia glathei at 1%, the degradation
remained unchanged for the other two isolates. A sudden
rise in degradation was seen at salinity of 1.5% with
Burkholderia glathei showing a significant rise of 24.99%,
while other two isolates showed an increase between 16 and
18 %. The results of the study indicate that the isolates used
in the study would be less effective bioremediating agents
in the marine environment. For further work, salinity was
maintained at 1.5%.
Nutrients, especially nitrogen, phosphorus and in some
cases iron are very important ingredients for successful bio-
degradation of hydrocarbon pollutants. Depending on the
nature of the impacted environment, some of these nutrients
could become limiting thus affecting the biodegradation
processes. This is more pronounced in marine environments
due to the low background levels of nitrogen and phospho-
rus in seawater unlike in freshwater systems, which regu-
larly fluctuate in nutrients as a result of perturbations and
receipt of industrial and domestic effluents and agricultural
runoff. The addition of nitrogen and phosphorus fertilizers
stimulates the biodegradation of hydrocarbons. It was re-
ported that the biodegradation rate of hydrocarbons is di-
rectly proportional to the supplemented nitrogen (Atlas 1981,
Cooney 1984). Leys et al. (2004) suggested a general theo-
 Varsha K. Vaidya and Sharmila Kadam
retically calculated C/N/P ratio of 100/10/1 (expressed in
moles) for hydrocarbon degradation. The minimal medium
used in the study contained 0.2g NH4Cl (54 mg nitrogen) as
the source of nitrogen, which was replaced by three differ-
ent sources of nitrogen viz., ammonium nitrate, urea and am-
monium chloride, added to represent nitrogen concentration
ranging from 5-50 mg N/100 mg naphthalene (93 mg C)
i.e., C to N ratios of 18.6:1 to 1.86:1 (Table 1). The highest
degradation was seen at C : N ratio of 9.3:1 with all the
sources of nitrogen, with urea proving to be the best source
of nitrogen showing an increase in degradation around 15%.
The overall trend showed a decrease in the degradation be-
low and above the optimum C : N ratio by all the isolates.
Bastiaens et al. (2000) showed that the type of nitrogen sup-
plement (i.e., ammonium versus nitrate) affected the degree
of oil biodegradation observed. Supplementation with am-
monium led to the progressive acidification of cultures,
whereas nitrate amendment resulted in a neutral culture me-
dium. In the present investigation, urea proved to be supe-
rior to both the sources of nitrogen.
A surprisingly large number of studies report no benefit
or even deleterious effects when excessive levels of N are
applied. Reports of specific inhibitory effects of excess N
include an increased lag phase and preferential inhibition of
aromatic degradation, although most indicate overall inhi-
bition of microbial respiration and/or hydrocarbon degrada-
tion. These factors could have contributed to lowered rates
of degradation at excessive concentrations of nitrogen in the
medium. Although nitrogen is considered to be the major
limiting nutrient, maximal hydrocarbon degradation occurs
with supplementation of both N and P (Braddock et al. 1997).
Feijoo-Siota et al. (2008) investigated the effect of avail-
ability of nitrogen and phosphorous on the uptake and deg-
radation of naphthalene by free cells in seawater collected
from a beach on the Galician coast (Lagos beach, Spain),
(pH 8.20, and salinity 33 mg/L). When necessary, the
seawater was supplemented with K2HPO4 as a source of phos-
phorous at a C : P ratio of 30:1, and NH4NO3 as a nitrogen
source, at a C : N ratio of 10:1. The results at 30°C revealed
that an initial naphthalene concentration of 25 mM was al-
most completely degraded (93%) within the first 6 days of
incubation in samples supplemented with nitrogen and phos-
phorous, whereas in the non-supplemented samples, only
42% of the naphthalene was consumed after that time. In
samples supplemented with nitrogen and phosphorous, 55%
the naphthalene was degraded, whereas in the non-supple-
mented samples the degradation was only 34%. Thus, sup-
plementing seawater with phosphorus and nitrogen was im-
portant in order to obtain the highest possible degree of bio-
degradation.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
In the present work, two sources of phosphorus were
employed viz., KH2PO4 and NaH2PO4 in ratios ranging from
0.07-0.12 mg P/100 mg naphthalene (C : P ratio ranging from
9.3:0.07 to 9.3:0.12) at C : N ratio (9.3:1) optimized in the
earlier step. Both the sources of phosphorus showed an en-
hanced degradation at a C : P ratio of 9.3:0.09 with potas-
sium di-hydrogen phosphate serving as the best source of
phosphorus for all three isolates. Thus, the optimum ratio
for the degradation was found to be 93/10/0.9. Thus, overall
addition of sources of nitrogen and phosphorus to the me-
dium resulted in an increase in the degradation of naphtha-
lene for all the isolates viz., Burkholderia glathei (from
40.56% to 54.24% leading to an increase of 33.72% in deg-
radation), Alcaligenes denitrificans (from 43.25% to 55.64%
leading to an increase of 28.65% in degradation), and Pseu-
domonas putida (from 45.42% to 56.63% leading to an in-
crease of 24.69% in degradation). The effect of nutrient sup-
plementation on Burkholderia glathei was thus more
pronounced.
PAHs present in the atmosphere or natural waters are
believed to be degraded to a limited extent by indigenous
hydroxyl radicals (OH·). The use of OH· generated by mixing
of H2O2 with FeSO4 i.e., Fenton’s reagent (5 mL 2.8M H2O2;
5 mL 0.1 M FeSO4) has shown promise for the degradation
of a wide range of environmental contaminants in aqueous
systems as well as soil slurry bioreactors. Fenton’s reagent
brings about oxidation of PAH’s to a more biodegradable
compound for further degradation by microorganisms.
The application of Fenton’s reagent to remediate matrices
contaminated by polycyclic aromatic hydrocarbons (PAHs)
shows that the matrix itself plays an important role, as the
iron oxides seem to be able to decompose hydrogen peroxide,
and thus initiate Fenton’s reaction. The PAHs sorbed onto
inner surface of the flask, and therefore not undergoing
oxidation, or PAHs sorbed on sewage sludges, contaminated
soils and sediments can be oxidized by the addition of
Fenton’s reagent (Martens & Frankenberger 1995). In the
present study, the effect of addition of Fenton’s reagent in
the aqueous system (MSM medium) under the conditions
optimized in the earlier steps was studied. Addition of
Fenton’s reagent enhanced the degradation of naphthalene
for all the isolates. The pronounced effect was seen in case
of Burkholderia glathei showing an increase in extent of
degradation by 10.33 % (59.85% degradation), while
Alcaligenes denitrificans and Pseudomonas putida showed
an increase of 10.84 % (61.68% degradation) and 9.58 %
(62.06% degradation) respectively.
Differences in the physical, chemical and microbial char-
acteristics of ecosystems can affect the disposition and per-
sistence of chemicals in the environment. Consideration of
 OPTIMIZATION OF CONDITIONS FOR NAPHTHALENE BIODEGRADATION 

these differences as well as the complexity of natural


ecosystems and the physical and chemical properties of

3HUFHQWGHJUDGDWLRQ


toxicants have led to the use of multicomponent microcosms

as laboratory models to predict the toxic effects and degra-
 %JODWKHL dation of chemicals in the environment (Giesy 1980). In the
$GHQLWULILFDQV
 present investigation the individual isolates and the consor-
3SXWLGD
 tia consisting of 1:1:1 mixture of the isolates was tested for
 their ability to degrade naphthalene in soil microcosms un-

der all the optimized conditions. The study was carried over

   
a period of seven days in soil previously equilibrated with
S+
naphthalene. The results of the present investigation indi-
Fig. 1: Effect of pH on degradation of naphthalene. cated the ability of all the isolates to establish themselves in
the soil. Pathak et al. (2008) carried out simulated micro-
cosm studies in the presence and absence of indigenous

 microflora confirmed its ability for naphthalene degradation
 and to colonize the soil. Pseudomonas sp. HOB1 was found
3HUFHQWGHJUDGDWLRQ

 to be highly potent in degrading higher concentrations of


 %JODWKHL naphthalene under laboratory conditions as well as in simu-
 $GHQLWULILFDQV lated microcosms, similar to results found in this study.
 3SXWLGD Heitkamp et al. (1987) studied naphthalene biodegradation


in microcosms containing sediment and water collected from
 three ecosystems which varied in past exposure to anthro-
      pogenic and petrogenic chemicals. Mineralization half-lives
3HUFHQWVDOLQLW\ for naphthalene in microcosms ranged from 2.4 weeks in
Fig 2: Effect of salinity on degradation of naphthalene. sediment chronically exposed to petroleum hydrocarbons to
4.4 weeks in sediment from a pristine environment. Mixed
microbial communities have the most powerful

biodegradative potential because the genetic information of

more than one organism is necessary to degrade the com-
3HUFHQW'HJUDGDWLRQ



3SXWLGD
plex mixtures of organic compounds present in the contami-

$GHQLWULILFDQV nated areas (Pumphrey et al. 2007). This was reflected in the

 %JODWKHL consortia proving to be superior to the individual isolates,
 FRQVRUWLD showing 89.15% degradation at the end of seven days (Fig.


3). Amongst the individual isolates Pseudomonas putida de-
 graded 76.27% of naphthalene at the end of seven days. The
      rate of degradation increased gradually over a period of seven
1XPEHURIGD\V
days, though the degradation observed at the end of 48 h
was lesser compared to the degradation in the liquid me-
Fig. 3: Degradation of naphthalene in soil microcosm. dium. The degradation observed at the end of 48 h in liquid
medium was nearly equivalent to degradation observed on

day five in the soil (58.56% for Burkholderia glathei, 62.18%
for Alcaligenes denitrificans and 65.46% for Pseudomonas

putida). The decreased degradation in soil can be contrib-
3HUFHQWGHJUDGDWLRQ


uted to the presence of organic matter in soil and sorption of
 3SXWLGD naphthalene to the soil making it unavailable to the degrad-
$GHQLWULILFDQV
 %JODWKHL ing organisms, despite the addition of Fenton’s reagent. The

&RQVRUWLD
degradation of chemicals in the environment can be affected
 by several factors which may differ among ecosystems, such
      as organic and inorganic nutrient levels, temperature, pH,
1XPEHURIGD\V
previous chemical exposure, microbial adaptations and oxy-
Fig 4: Effect of immobilization of cells on degradation of naphthalene. gen tension. In soil, PAHs are heterogeneously distributed

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Varsha K. Vaidya and Sharmila Kadam

Table 1: Effect of addition of nitrogen source on naphthalene biodegradation.

Nitrogen source mg ofnitrogen/100 mg Percent degradation

of naphthalene (C:N) Burkholderia glathei Alcaligenes denitrificans Pseudomonas putida

NH4NO3 5 (18.6:1) 43.50 ± 1.10 45.00 ± 1.10 46.36 ± 1.12

10 (9.3:1) 48.20 ± 1.18 49.10 ± 1.03 52.35 ± 1.16
15 (6.2:1) 46.30 ± 1.05 47.80 ± 0.98 50.21 ± 1.21
20 (4.65:1) 38.50 ± 1.00 39.10 ± 0.79 42.18 ± 1.12
25 (3.72:1) 36.80 ± 0.95 37.00 ± 0.84 39.15 ± 0.93
50 (1.86:1) 33.90 ± 0.88 35.80 ± 0.98 37.64 ± 0.84
Urea 5 (18.6:1) 45.60± 1.01 46.90 ± 1.06 48.50 ± 1.15
10 (9.3:1) 49.70 ± 1.16 51.50 ± 1.22 54.00 ± 1.18
15 (6.2:1) 47.30 ± 1.13 49.36 ± 0.93 52.10 ± 1.15
20 (4.65:1) 46.10 ± 1.11 47.90 ± 1.08 48.20 ± 1.12
25 (3.72:1) 45.23 ± 1.08 45.25 ± 1.09 46.90 ± 1.15
50 (1.86:1) 41.16 ± 1.00 44.18 ± 1.10 45.20 ± 1.05
NH4Cl 5 (18.6:1) 42.16 ± 0.97 44.20 ± 1.05 51.20 ± 1.07
10 (9.3:1) 46.28 ± 1.02 48.12 ± 1.25 52.00 ± 1.12
15 (6.2:1) 45.60 ± 1.01 46.82 ± 1.16 48.90 ± 1.02
20 (4.65:1) 43.12 ± 1.01 44.25 ± 1.04 45.60 ± 1.00
25 (3.72:1) 42.74 ± 0.88 43.26 ± 1.02 45.60 ± 1.01
50 (1.86:1) 40.20 ± 1.00 43.25 ± 0.97 45.20 ± 1.13

Values in parentheses indicate C: N ratios.

Table 2: Effect of addition of phosphorus source on naphthalene biodegradation

Source of mg of phosphorus/100 mg Percent degradation

Phosphorus of naphthalene Burkholderia glathei Alcaligenes denitrificans Pseudomonas putida

KH2PO4 0.07 (93:0.7) 46.56 ± 1.10 48.12 ± 1.13 49.96 ± 1.04

0.08 (93:0.8) 51.96 ± 1.28 53.45 ± 1.05 54.24 ± 1.17
0.09 (93:0.9) 54.24 ± 1.17 55.64 ± 1.22 56.63 ± 1.12
0.10 (93:1) 46.65 ± 1.00 47.96 ± 1.14 49.30 ± 1.09
0.11 (93:1.1) 45.20 ± 1.09 46.12 ± 1.00 48.30 ± 1.00
0.12 (93:1.2) 42.56 ± 1.01 43.21 ± 1.04 47.00 ± 1.05
Na2HPO4 0.07 (93:0.7) 45.56 ± 1.16 46.23 ± 1.16 47.56 ± 1.06
0.08 (93:0.8) 51.96 ± 1.14 51.24 ± 1.18 52.13 ±1.23
0.09 (93:0.9) 54.21 ± 1.21 53.14 ± 1.23 54.32 ± 1.18
0.10 (93:1) 46.65 ± 1.01 46.20 ± 1.15 47.30 ± 1.15
0.11 (93:1.1) 45.20 ± 1.06 44.30 ± 1.04 46.50 ± 1.07
0.12 (93:1.2) 42.56 ± 1.02 42.30 ± 1.01 45.36 ± 1.03

Values in parentheses indicate C:P ratios.

and may be absorbed inside of organic particles, located in
small pores that are inaccessible for bacteria, or otherwise
occluded by the multitude of solid soil constituents
(Heitkamp et al. 1987). The results obtained in the present
study, however, showed between 8.46 % and 14.86 % deg-
radation at the end of 24 h.
Treating large amounts of hydrocarbon contaminated
liquid or soil in bioreactors is technically challenging and
costly. Bioaugmentation (i.e. seeding the soil with
hydrocarbon-degrading microorganisms such as fungal and/
or bacterial cells) can circumvent many technical difficulties,
but to date releasing free cells into a polluted environment
has produced variable results. Failure of hydrocarbon
degradation has often been attributed to the inability of the
introduced bacteria to compete with the indigenous
microorganisms that are better adapted to the rapidly
changing environmental conditions (Alexander 1999).
Therefore, it is suggested that immobilizing the bacteria in
non-liquid inocula can achieve more homogenous conditions
and better protection of the cells. An effective approach to
finding a solution to continuous contamination is to
concentrate these biodegraders on the natural or synthetic
supports with high porosity thus allowing their use in
different kinds of bioreactors (fluidized bed, packed bed).
Immobilization presents many advantages such as the
presence of high population density in a limited reactor
volume, high conversion rates, limiting substrate inhibition
and toxicity to micro-organisms by diffusional limitations,
reusability and reduction in the overall cost (Trevor & Elsas
1992). Immobilization would also have the added benefit of

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 OPTIMIZATION OF CONDITIONS FOR NAPHTHALENE BIODEGRADATION
reducing the cells’ exposure to the elements, thus preventing
them from being washed away, particularly in an open water
system. Immobilized materials, furthermore, have
comparatively longer operating lifetimes due to an enhanced
stability of the macromolecules or cells and, consequentially,
protection from adverse conditions. Various methods have
been described for the immobilization of bacteria and
entrapment within polymeric gel matrices has often been
successful. The most frequently used matrices are N-
carrageenan, alginate and polyacrylamide. Alginate,
however, represents several advantages such as high porosity,
chemical stability and low cost. It is a mild, fast and simple
immobilization method. Diaz and co-workers (2001)
reported that immobilization of bacterial cells significantly
enhanced the biodegradation rate of crude oil compared to
free-living cells over a wide range of culture medium salinity.
Immobilizing agents offer cellular protection against adverse
environmental conditions. In the present investigation, the
three naphthalene degraders along with the consortia were
immobilized and their ability to degrade naphthalene was
followed over a period of seven days (1, 2, 3, 5 and 7 days).
The rate of degradation increased steeply on day two
followed by a gradual increase thereafter (Fig. 4). The
immobilized cells degraded naphthalene to a greater extent
compared to the free cells. Immobilized Burkholderia glathei
showed an increase of 5.09 % in the rate of degradation, while
Alcaligenes denitrificans and Pseudomonas putida showed
an increase of 4.67 % and 7.46% respectively at the end of
48 h. Consortia again proved to be superior to the individual
isolates showing 98% degradation of naphthalene at the end
of seven days as compared to the individual isolates viz.,
Burkholderia glathei (71.16%), Alcaligenes denitrificans
(76.25%) and Pseudomonas putida (89.12%). The effect of
storage on the viability of the immobilized cells (initial
inoculum 1 × 108 cells/mL) after storing for 15 days at 4°C
was analysed and compared to the viability of freshly-made
beads. The results showed no appreciable loss of viability
indicating that storage at 4°C for 15 days did not affect the
viability of the immobilized cells adversely. In a similar study
Feijoo-Siota et al. (2008), showed that the immobilized cells
can be stored at 4°C for at least one month with no
appreciable loss of viability, whereas storage at -20°C was
not as suitable. This is advantageous for practical use of the
beads, because it shows that they do not require any special
or expensive storage conditions.
The results of this work indicate that bioremediation
technology has an immense potential for decontamination
of polluted natural environments and concludes that
successful application of bioremediation depends on
hydrocarbon degrading microorganisms and in situ
environmental conditions.
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

5()(5(1&(6
Aislabie, J., Fraser, R., Duncan, S. and Farrell, R.L. 2006. Effects of oil
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Kailas F. Khairnar, S. T. Ingle, Nilesh D. Wagh and Lalit T. Ingale
School of Environmental and Earth Sciences, North Maharashtra University, Umavi Nagar, Jalgaon-425 001, Maharashtra

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,1752'8&7,21 age to lung tissue, carcinogenesis and premature death (Cotes

The issue of transportation and the environment is paradoxi-
cal in nature. From one side, transportation activities sup-
port increasing mobility demands for passengers and freight,
and this ranging from urban areas to international trade. On
the other side, transport activities have resulted in growing
levels of motorization and congestion. As a result, the trans-
portation sector is becoming increasingly linked to environ-
mental problems. The relationships between transport and
the environment are multidimensional. The most important
impacts of transport on the environment relate to climate
change, air quality, noise, water and soil quality as well as
loss of biodiversity. Transport systems becomes nuisance to
the public when they pass through or near the residential
areas (Carpenter 1994). Transport sector encompasses high-
way vehicles, marine engines, locomotives and aircraft,
which are the sources of pollution in the form of gases and
particulate matter emissions that affects on air quality caus-
ing damage to human health. Ambient air is most polluted
in cities, with the increase in the number of motor vehicles
caused by economic growth and industrialization, the level
of pollution is expected to worsen further in the future (Alam
et al. 1999). Long term exposure to air pollution is an im-
portant factor in the development of chronic respiratory dis-
eases (Karakatatsani et al. 2003). The effects of air pollution
include breathing and respiratory problems, aggravation of
existing respiratory and cardiovascular diseases, alteration
in the body defence systems against foreign materials, dam-
1978, NHLBI/WHO 1995).
The particles emitted from the exhaust of more than 10
micron size are held in upper respiratory tract and particles
less than 10 micron size (PM10) accumulate in lungs and pro-
duce respiratory abnormalities. Hence, PM10 are of great con-
cern in air pollution studies (Ingle et al. 2005). Long term
exposure to particulate matter for years or decades is associ-
ated with elevated cardiovascular problems, infant mortal-
ity and morbidity, respiratory symptoms, and effects on lung
growth and function of immune system. Short term study
show consistent association of exposure to daily concentra-
tions of PM with mortality and morbidity on same day or
subsequent days. The major subgroups of the population that
appear to be most sensitive to the effects of particulate mat-
ter include individuals with chronic obstructive pulmonary,
cardiovascular disease, influenza, pneumonia and other res-
piratory diseases and asthma (Kappos et al. 2004, Balmes
1993).
Railway is an important means of transport in India. In-
dian Railways has one of the largest and busiest rail net-
works in the world, transporting over 18 million passengers
and more than 2 million tonnes of freight daily (Indian Rail-
ways Year Book 2007, Rao 2008). The emissions from rail
activities are directly related to the construction during the
completion of the foundation work, superstructure and track
laying, signal and telephone lining, and electrical lining
(Banverket & Luleå 2006) before implementation of regular
 Kailas F. Khairnar et al.
railway activity. Coal generated smoke in steam days was a
significant part of atmospheric pollution. Although steam
traction is now only of historic interest due to introduction
of electric traction, it still has a part to play in the areas where
electrification has not been entirely phased in. Steam loco-
motives emit smoke particulates in the vicinity and contrib-
ute sulphur and nitrogen oxides and CO2 to the wider at-
mosphere. Diesel locomotives and rail motor units produce
the same sort of emissions as do road lorries, including car-
bon monoxide, nitrogen oxides, hydrocarbon and carbon
based particulates. The contribution of electric transport to
atmospheric pollution occurs where electricity is generated
at fossil fuel burning power stations (Carpenter 1994). The
sources of pollution in railway stations are welding fumes
and iron particulate emission from rail track due to speed of
train. People are exposed to iron oxide, assuming the fine
iron abrasion dust is rapidly oxidized, with trace amounts of
chromium, copper, zinc, manganese, and quartz. These cases
are mostly observed in underground railways (Seaton et al.
2005). Godowns located on or nearby areas of railway sta-
tions are also the source of dust emission in railway stations.
Varieties of goods carried out through freights are emptied
in the godowns for its consecutive journey. Particulate mat-
ter generated by these goods is added in the atmosphere dur-
ing its handling, which acts as a source for ambient air pol-
lution on platform. Renovation activities in the railway sta-
tion or nearby areas, which include breaking of old construc-
tions and construction of new structures, are the source of
dust pollution in railway stations.
The volume and spatial distribution of the emissions, as
well as dispersion conditions, affect pollution levels. Sev-
eral other factors also play a part in determining the expo-
sure of a population. Pollution intake is also determined by
the number of people in polluted areas, how long they stay
there and what they do (Krzyzanowski, WHO 2005). Time-
activity patterns, particularly residence or work near busy
stations (or both), and time spent in traffic are critical for
population exposure. Population growth and future expan-
sion of urban centers are not considered while planning of
rail projects in India. As a result large proportion of popula-
tion is exposed to railway transportation (Khairnar & Ingle
2009).
Occupational activities, which involve exposure to dust
and plant source particulate matter affect the lung capacity
and cardio-respiratory fitness of workers (Debray et al. 2002).
The impairment in lung efficiency increases with duration
of exposure in workers (Wagh et al. 2006). Shopkeepers work
at high risk of exposure to the air pollution (Ingle et al. 2005).
Shop assistants in an air conditioned environment should be
less exposed to traffic fumes and their lung should be better
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
preserved compared to unprotected vendors exposed directly
to vehicular pollution (Jones et al. 2008). Travelers are of-
ten exposed to levels that are three times the background
levels. Groups with high levels of exposure include people
who live near transportation activity and people whose jobs
require them to spend a long time on the stations. Urban
planning and development also strongly shape exposure; they
determine not only patterns of residence and mobility but
also the availability of public transport.
The present study emphasizes on the health effects, es-
pecially pulmonary function test and ventillary impairment,
among population exposed to railway transportation in
Jalgaon region. The Jalgaon railway shares 350 km from
5,440 km of total railway network in Maharashtra. Bhusawal
railway division is the Central Railway’s one of the most
important headquarters located at 24 km from Jalgaon. The
rail route passes through Jalgaon covering approximately 8
km distance through city. The residential colonies are well
developed on both sides of railway track. Many vendors are
also established their business on both sides of the track.
0$7(5,$/6$1'0(7+2'6
Site description: Jalgaon and Bhusawal railway stations
were selected for the present study. Jalgaon is a city in west-
ern India, to the north of Maharashtra state. Jalgaon is one of
the fastest developing cities in north region of the state (Wagh
et al. 2003). Jalgaon railway station is a junction for the trains
coming from Central and Western Railways. Bhusawal, is
located on the bank of Tapi river, and is biggest taluka of
Jalgaon district. Bhusawal has very good railway connec-
tivity, and an important divisional headquarters of Indian
Central Railway. Workplaces of IWO (Inspection of Works),
C&W (Carriage & Wagons), RPF (Railway Police Force)
and Operating Department along with workplace of coolie
of stations, residential areas and shops located on both sides
of railway track passing through the Jalgaon city were se-
lected for the study purpose.
Study population: Working population of IWO, C&W,
RPF, Operating Department along with licensed Coolie,
Population doing daily up-down from same stations were
selected as subjects for the study purpose. Residential peo-
ple living either sides of track and shopkeepers of shops lo-
cated around railway activity were also considered as sub-
jects. The control subjects were selected from general popu-
lation of the Jalgaon city working in Banks or Government
offices. It was ensured that these subjects are not exposed to
any type of air pollution. Subjects having asthmatic history
were rejected from the study. Number of exposed subjects
was 50 each, and control subjects 60 respectively.
 RAILWAY TRAFFIC POLLUTION AND HEALTH EFFECTS
Screening questionnaires: The data on the health status of
the study groups were collected using the standard Respira-
tor Medical Evaluation Questionnaire (OSHA 1998) trans-
lated into a local language.
Workplace environment: The work was carried out in
March-April. The humidity of workplace environment of
railway employees and indoor environment of exposed popu-
lation varied from 46% to 53.5%, while temperature was in
the range of 30.5°C to 40.5°C.
Dust exposure monitoring: The exposure of dust was meas-
ured by a portable dust sampler over an 8-h period. The sam-
pling unit contained an air pump powered by an internally
sealed lead-acid gel battery. Air was drawn at a flow rate of
0.5 to 3.5 litres per minute, and the dust sampling unit was
attached to the body of the object for 8-h per day. The dust
(PM10) was collected by filtration of air through glass fibre
filter (25 mm diameter). The samples collected were meas-
ured by the gravimetric method and expressed as PM10 dose
in µg/m3.
Pulmonary function test: The target groups and control
samples were subjected to the Pulmonary function test
(Williams 1986, Jeelani 1992, Krelt et al. 1989). The pul-
monary function tests were conducted by the instrument
Spirometer (Medspiror Recorder & Medicare Systems
Chandigarh, India). The Medspiror is based on the volume
differential method for flow detection. Forced vital capacity
(FVC) is the maximal amount of air that can be exhaled fol-
lowing maximal inspiratory effort (Normal: Male 4.8 L,
Female 3.1 L). Forced expiratory volume (FEV1) is the vol-
ume of air exhaled in one second during a forced vital ca-
pacity effort and peak expiratory flow rate (PEFR) is the
maximum amount of air exhaled with forced effort during
the FVC. Before spirometric test of the subjects, their age,
height, weight and gender were fed into the spirometer.
Spirometer gives two values viz., predicted and actual. The
predicted values are based on the age, height, weight and
gender whereas the actual values are based on the maximal
exhalation followed by maximal inspiration of the subject.
The pulmonary function test was conducted by sitting the
subject comfortably in a chair for clearance manoeuvre (Badr
et al. 2002). Regular sterilization of mouthpieces was done
before the use. The subjects were asked for maximum exha-
lation followed by maximum inspiration. Three such tests
were performed and subjects were asked to improve the per-
formance. Best of three performances of FVC, FEV1 and
PEFR were taken into consideration. The equations for pre-
diction were as follows.
FVC (L) = 0.050H – 0.014A – 4.49
FEV1 (L) = 0.040H – 0.021A – 3.13
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

PEFR (L /Sec) = 0.071H – 0.035A – 1.82
Where, H is height in cm and A is age in years.
The results of spirometry were assessed as per the crite-
ria given in the manual of the Medspiror. The Medspiror
software using a set of prediction equations for the adults
calculates the expected values (Krelt et al. 1989).
The tests were conducted in the morning hours and en-
sured that the subjects were not exposed to air pollution at
least for 12 hours before the test (Pandey 2001).
Statistical analysis: In the present study, some simple sta-
tistical parameters were applied with advanced statistical
parameters such as Analysis of Variance (ANOVA) to the
collected data. The data were processed for mean, standard
deviation and two-way ANOVA (Armitage & Berry 1994).
It comprises recording of FVC, FEV1 and PEFR.
Risk assessment: The data on health status of the study
groups were collected by standard questionnaire. The Res-
pirator Medical Evaluation Questionnaire (OSHA 1998, ISO-
7708, 1995) was used for collection of the data. The symp-
toms viz., frequent coughing, shortness of breath and irrita-
tion in respiratory tract were considered for risk analysis.
The risk (Ito & Thurston 1996, Sengupta 1996) were calcu-
lated for the target groups (IWO, C&W, RPF, Operating
Department, Coolie, Residential, Shopkeepers and Up-down-
ers) against control group having different exposure to risk
factors. The odds ratio, relative risk and attributable risk were
calculated by setting a simple 2 × 2 matrix (Gilbert 2004)
such that the rows divide the population according to those
who had been exposed and those who have not been exposed
(control) to the risk factor. According to Hennekens & Buring
(1987), odd ratio is nothing but the odds of disease in ex-
posed persons divided by odds of disease in unexposed per-
sons. Attributable risk is, finding the probability of disease
for exposing to the particular factor. Sackett et al. (1996)
reported that the attributable risk is a measure of excess risk
accounted for by exposure to a particular factor. Relative
risk is the ratio of two absolute risks. It measures the strength
of effect of an exposure on risk (Sackett et al. 1996). The
columns were based on the number of individuals who had
acquired the symptoms being studied and those who had not
in both of the target groups.
5(68/76
Environmental monitoring of exposed and control
groups: Table 1 shows the environmental monitoring of each
group. The average temperature of IWO, C&W, RPF, oper-
ating, Coolie, Up-Down, Residential, Shopkeepers and Con-
trol groups during study period were 34°C, 40.5°C, 37.5°C,
31°C, 39.5°C, 39°C, 32.5°C, 33.5°C and 30.5°C respectively,
 Kailas F. Khairnar et al.
while the relative humidity of the same places was 49.5%,
46 %, 49%, 53%, 49%, 48%, 53.5%, 46.5% and 49.5% re-
spectively. Air monitoring of these places shows PM10 con-
centration of 366.37, 460.52, 206.76, 182.73, 394.73, 308.02,
174.36 and 293.21 respectively. Each group is classified as
indoor, mix and outdoor. IWO and Shopkeepers are the
groups working in mix environment whereas Residential,
operating and C&W are categorized in indoor environment.
Remaining groups i. e., Coolie, R.P.F. and Up-downers
groups are categorized in outdoor environment.
Pulmonary status of exposed and control groups: Table 2
shows the physical parameters of the subjects i. e., age, height
and weight, which were considered for the spirometric test.
The age of subjects selected for the study ranged between 18
and 53 years. Average weight of the subjects was ranged
from 24 kg to 116 kg. The average height of the respondents
was 152 cm and ranged from 131 to 187cm.
Observed values and percentage of pulmonary function
tests show decline in the FVC, FEV1 and PEFR indices of
the exposed population as compared to the control group.
Average value of FVC in IWO, C&W, RPF, operating,
Coolie, Up-Down, Residential, Shopkeepers and control
groups was 76%, 83%, 83%, 77%, 75%, 79%, 73%, 77%
and 92% respectively. The average value of FEV1 in the same
groups was 86%, 96%, 92%, 87%, 80%, 85%, 82%, 83%
and 101%. Peak Expiratory Flow Rate (PEFR) is the best
test of expiratory effort. Its values in the exposed and con-
trol groups were 69%, 69%, 79%, 72%, 66%, 76%, 73%,
62% and 84% respectively.
Ventillary impairment in target groups: The ventillary
impairment of exposed group is categorized on the basis of
air flow obstruction (FVC), restrictive defect (FEV1) and
expiratory flow rate (PEFR) in as given in Table 3. The data
show that all the groups have warning signal and fatal asthma.
Coolie and IWO workers are more affected groups having
32% and 20% fatal asthma respectively, whereas operating
and shopkeepers have 62% and 74% warning signals. All
the groups show stable asthma less than 50%. Control group
shows 65% stable asthma. Severe restrictive defect is ob-
served in Coolie (10%), Up-downers (2%), Residential (2%)
and Shopkeepers (2%). All the groups except C&W shows
moderate restrictive defect. Mild restrictive defect is observed
in all groups, whereas it is more in IWO (32%), Up-down-
ers (34%) and Residential (36%) groups. Control group
shows no severe and moderate effect. Severe air flow ob-
struction is observed in IWO (2%), Coolie (10%), Up-down-
ers (2%), Residential (8%) and Shopkeepers (4%). Moder-
ate air flow obstruction is observed in each group, whereas
mild air obstruction is observed more in Up-downers (60%)
group. Control shows no severe and moderate air flow ob-
struction effects.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
Risk assessment of population exposed to workplace
environment: Table 4 shows the higher risk for all the groups
exposed to the ambient air prevailing at the workplace envi-
ronment. Relative risk and odd ratio for the symptoms stud-
ied were above one in all the exposed groups, which indi-
cates an association between exposure and group. Coolies
are on higher risk having relative risk of 2.58, 6.33 and 6.5,
whereas odd ratio 3.84, 9.6 and 8.43 for symptoms like fre-
quent coughing, shortness of breath and irritation in respira-
tory tract respectively. Attributable risk in all the groups
shows strong relationship in exposure and groups.
Pulmonary status (as per years of exposure) of exposed
and control groups: All the groups were categorized into 5
categories as per their duration of exposure. Test was per-
formed for comparison of individual pulmonary function test
of IWO, C&W, RPF, operating, Coolie, UP-Down, Residen-
tial, and Shopkeeper against control group. All the groups
are categorized according to 0 to 5, 6 to 11, 12 to 16, 17 to
22 and above 22 years of exposure as in Tables 5, 6, 7,8 and
9 respectively, where data show that p values in ANOVA
are significant p<0.05 by multiple comparison tests. The
comparison of duration of dust exposure with pulmonary
function test of each individual group against control group
clearly indicates a relation between dust exposure and pul-
monary functions in exposed groups.
',6&866,21
Temperature of exposed groups was in the range of 30.5°C
to 40.5° C. Workplace temperature of Coolie and C&W group
is more as they are working on platform (outdoor) and in the
carriage and wagons (indoor) respectively. Observed value
of PM10 concentration of C&W and Coolies was more. Pas-
senger trains, freights containing variety of goods like coal,
food items, agriculture products, fertilizers, etc. are carried
out from the stations. Particles of these goods sometimes
mix with the atmosphere and increase the concentration of
particulate matter on the stations. Godowns, which are lo-
cated on Bhusawal as well as on Jalgaon railway stations,
are also the source of dust emission in ambient atmosphere
while handling or transferring the goods. Due to speed of
trains dust from surroundings is carried out with the flow
when trains enter the stations. At the same time the coolies
become exposed population to this dust who are waiting for
the customers on platform. C&W workers work in carriage
and wagons which are standing on platforms, so they are
also exposed to heavier concentration of dust. Whenever
renovation activity is conducted on Bhusawal station, it con-
tributes to the source of dust pollution on platform. FVC of
all the groups has declined as compared to control group,
where group of coolie and residential show less FVC i. e.,
75% and 73% respectively. It may be due to accumulation
 RAILWAY TRAFFIC POLLUTION AND HEALTH EFFECTS 

Table1: Environmental monitoring of IWO, C&W, RPF, Operating Department, Coolie, Up-Downers, Residential, Shopkeepers and Control Samples.

Sr No. Target PM10 dose Average Relative Type

Groups (µg/m3) Temperature °C Humidity, %

1 IWO 366.37±71.11 (316.09-416.6) 34 ± 2.82 (32-36) 49.5 ± 3.53 (47-52) Mix

2 C&W 460.52 ± 62.02 (416.6-504.38) 40.5 ± 3.53 (38-43) 46 ± 4.24 (43-49) Indoor
3 RPF 206.76±44.30 (175.43-244.25) 37.5 ± 4.94 (34-41) 49 ± 8.48 (43-55) Outdoor
4 Operating Deptt. 182.73±87 (121.21-244.25) 31 ± 4.24 (28-34) 53 ± 5.65 (49-57) Indoor
5 Coolie 394.73 ± 31.01 (372.8-416.6) 39.5 ± 6.36 (35-44) 49 ± 8.48 (43-55) Outdoor
6 Up-Downers 308.02±8.91 (301.72-314.32) 39 ± 4.24 (36-42) 48 ± 7.07 (43-53) Outdoor
7 Residential 174.36±73.87 (122.12-226.60) 32.5 ± 4.94 (29-36) 53.5 ± 7.7 (48-59) Indoor
8 Shopkeepers 293.21 ± 42.5 (263.15-323.27) 33.5 ± 6.36 (29-38) 46.5 ± 7.77 (41-52) Mix
9 Control NA 30.5 ± 4.94 (27-34) 49.5 ± 10.6 (42-57) Indoor

Table 2: Pulmonary status of different exposed and Control groups.

Sr.Parameter A (n=52) B (n=50) C (n=50) D (n=50) E (n=50) F (n=49) G (n=52) H (n=50) I (n=59)
No.

1 Age (yr) 41.16±7.59 40.14 ± 8.52 39.8 ± 9.02 37.32±9.42 37.04 ±9.24 29.6±5.79 33.0 ±10.90 35.88±9.87 40.52±7.27
(24-52) (24-51) (20-51) (18-51) (21-51) (19-46) (19-51) (20-51) (27-51)
2 Height 161.58 ± 7.68 161.94± 8.53 165.26 ± 7.42 166.42 ± 7.65 163.8 ± 5.92 164.2± 9.87 159.89± 9.82 164 ± 9.01 161.83±15.04
(cm) (143-177) (145-180) (150-179) (153-186) (148-175) (140-181) (132-180) (143-176) (140-181)
3 Weight 60.06±10.42 63.62±7.95 66.48±12.72 63.18±13.44 58.24±8.68 57.96±7.49 56.21±13.07 60.06±12.89 65.30±8.27
(kg) (40-86) (45-84) (41-95) (44-116) (44-82) (48-86) (25-82) (35-86) (45-82)
4 FVC Obs 2.15±0.85 2.43±0.68 2.54±0.81 2.52±0.64 2.27±0.82 2.46±0.63 2.13±0.66 2.44±0.86 7.53±35.82
(L) % 76 83 83 77 75 79 73 77 92
5 FEV1 Obs 2.0±0.66 2.32±0.58 2.28±0.62 2.33±0.60 2.02±0.80 2.21±0.55 1.97±0.58 2.17±0.74 2.54±0.51
(L) % 86 96 92 87 80 85 82 83 101
6 PEFR Obs 5.46±1.70 5.56±1.62 6.08±1.72 6.13±1.79 5.44±1.99 8.03±13.35 7.36±1.90 5.18±1.70 6.78±1.48
(L/s) % 69 69 79 72 66 76 73 62 84
7 FEV1/ Obs 92.0±2.10 93.80±8.48 91.79±12.13 92.02±16.02 5.44±1.99 93.60±14.43 91.92±10.79 88.59±16.63 89.54±12.54
FVC (%) % 107 114 114 113 106 112 111 107 111

A = IWO, B = C&W, C = RPF, D = Operating Department, E = Coolie, F = UP-Downers, G = Residential, H = Shopkeeper, I = Control

Table 3: Ventillary impairment in target groups.

Sr. No. Lung status A B C D E F G H I

1 Air flow obstruction

Normal (FVC > 80 %) 34 46 52 44 42 32 34 42 91.5
Mild (FVC 60 - 80 %) 36 46 40 40 30 60 42 36 8.4
Moderate (FVC 40 - 60 %) 28 8 8 16 18 6 20 16 0
Severe (FVC < 40 %) 2 0 0 0 10 2 8 4 0
2 Restrictive defect
Normal (FEV1 > 80 %) 52 86 70 62 54 56 42 56 93.3
Mild (FEV1 60 - 80 %) 32 14 28 28 20 34 36 28 6.7
Moderate (FEV1 40 - 60 %) 16 0 2 10 16 6 14 12 0
Severe (FEV1 < 40 %) 0 0 0 0 10 2 2 2 0
3 Asthma symptoms
Stable asthma (PEFR > 80 %) 30 28 42 28 26 38 38 12 64.04
Warning signal (PEFR 50 - 80 %) 50 54 52 62 42 52 50 74 32.2
Fatal asthma (PEFR < 50 %) 20 18 6 10 32 10 16 14 5.08

A = IWO, B = C&W, C = RPF, D = Operating Department, E = Coolie, F = Up-Downers, G = Residential, H = Shopkeeper, I = Control

of dust particles in the lung airways of both the groups as
these groups are more exposed to the dust due to workplace
environment of platform and residential group which is lo-
cated on either sides of track. FEV1 of all the groups shows
decline compared to the control group but coolies are more
affected. PEFR is the parameter which is adversely affected
in shopkeepers and coolie i. e., 62% and 66% respectively
among all the exposed groups. RPF is the group which is
least affected in all the groups with respect to FVC, FEV1
and PEFR parameters. It may be due to their workplace ex-
posure of dust which is very less in all the groups. The healthy
physical condition of RPF group is also responsible for fewer

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 Kailas F. Khairnar et al.

Table 4: Risk assessment of population exposed to workplace environment.

Sr. Target Risk Assessment

No. Group Frequent coughing Shortness of Breathe Irritation in respiratory tract
Relative Attributable Odd Relative Attributable Odd Relative Attributable Odd
risk risk Ratio risk risk Ratio risk risk Ratio

1 IWO 1.88 0.15 2.3 3 0.12 3.44 3 0.28 3.27

2 C&W 2 0.17 2.52 3.16 0.13 3.67 4.25 0.23 4.92
3 RPF 1.35 0.06 1.46 2 0.06 2.14 2.75 0.07 2.97
4 Operating Deptt. 1.47 0.08 1.63 2.16 0.07 2.34 2.25 0.05 2.37
5 Coolie 2.58 0.27 3.84 6.33 0.32 9.6 6.5 0.22 8.43
6 Up-Downers 1.05 0.01 1.07 2.16 0.07 2.34 2.75 0.07 2.97
7 Residential 1.05 0.01 1.07 2 0.06 2.14 2 0.04 2.09
8 Shopkeepers 1.88 0.15 2.3 4 0.18 4.95 4.75 0.15 5.63

a
Relative risk and odds ratio above 1.0 indicates an association between exposure and risk.
b
An attributable risk of 0.0 suggests no relationship, and above it strong relationship.

Table 5: Pulmonary status of different exposed and Control groups (0 to 5 yrs exposure).

Sr. T.G. 0 to 5 yrs Exposure

No. FVC (L) FEV1 (L) PEFR (L/s) FEV1/FVC (%)
Obs % Obs % Obs % Obs % p-value

1 A (n=8) 2.05 ± 0.65 68.10 ±29.00 1.97 ± 0.73 76.35 ±32.27 5.93 ± 1.79 71.70 ± 22.19 94.29 ± 14.83 107.63 ±14.52 <0.0001
2 B (n=11) 2.98 ± 0.84 99.00 ±27.74 2.56 ± 0.66 100.78 ±26.44 5.08 ± 1.5 61.72 ±16.67 88.91± 11.85 103.27 ±12.02 <0.0001
3 C (n=8) 2.43 ± 0.68 81.27 ± 16.67 2.35 ± 0.71 93.62 ± 20.06 6.22 ± 1.85 79.64 ±13.28 96.24 ± 8.55 114.83 ± 9.54 <0.0001
4 D (n=12) 2.72 ± 0.74 74.11± 16.47 2.49 ± 0.77 79.55 ± 18.11 6.23 ± 1.84 67.49 ±18.24 85.63 ± 29.71 100.25 ± 20.83 <0.0001
5 E (n=12) 2.52 ± 0.92 78.01 ± 27.37 2.52 ± 0.80 86.89 ± 28.53 6.73 ± 1.92 76.65 ±23.45 89.99 ± 14.90 102.59 ± 13.73 <0.0001
6 F (n=39) 2.53 ± 0.63 79.31 ± 20.74 2.25 ± 0.55 84.26 ± 18.12 8.71 ± 15.04 85.14 ±19.08 93.68 ± 15.99 112.98 ± 10.31 <0.0001
7 G (n=11) 2.18 ± 0.69 74.65 ± 18.66 1.90 ± 0.59 78.18 ± 21.55 4.56 ± 1.80 59.76 ±18.93 90.48 ±18.35 109.23 ± 23.25 <0.0001
8 H (n=18) 2.88 ± 0.77 80.44 ± 19.43 2.68 ± 0.63 87.29 ± 19.82 6.15 ± 1.02 66.12 ±10.77 94.10 ± 5.69 109.21 ± 6.40 <0.0001
9 I (n=10) 3.37 ± 0.53 101.3 ± 8059 2.84 ± 0.69 103.64 ± 16.31 6.82 ± 1.28 81.28 ±15.26 84.798 ± 17.4 103.54 ±14.13 <0.0001

Overall difference is based on two-way ANOVA. Test was performed for comparison of individual pulmonary function test of IWO, C&W, RPF,
Operating Deptt., Coolie, Up-Downers, Residential, Shopkeeper against Control group, where p values in ANOVA are significant p<0.05 by multiple
comparison tests. A = IWO, B = C&W, C = RPF, D = Operating Deptt., E = Coolie, F = Up-Downers, G = Residential, H = Shopkeeper, I = Control
Table 6: Pulmonary status of different exposed and Control groups (6 to 11 yrs exposure).

Sr. T.G. 6 to 11 years exposure

No. FVC (L) FEV1 (L) PEFR (L/s) FEV1/FVC (%) p-value
Obs % Obs % Obs % Obs %

1 A (n=11) 1.66 ±0.49 64.34 ± 23.45 1.88 ±0.84 81.38 ± 36.94 4.41 ± 1.79 59.59 ± 20.44 93.77 ± 10.30 112.94 ± 29.74 <0.0001
2 B (n=7) 2.32±1.01 82.85 ± 26.62 2.74±0.81 103 ± 35.22 5.38 ± 2.06 65.85 ± 23.38 95.89 ± 7.27 115.40 ± 7.364 <0.0001
3 C (n=5) 3.20±0.70 101.26 ± 13.28 2.58±0.55 98.47 ± 19.56 5.66 ± 1.37 68.68 ± 11.9 79.84 ± 20.42 98.81 ± 25.2 <0.0001
4 D (n=9) 2.37±0.63 73.60 ± 19.6 2.26±0.59 84.32 ± 21.66 6.38 ± 2.00 76.77 ± 20.32 96.08 ± 7.73 115.71 ± 10.80 <0.0001
5 E (n=4) 2.28±0.44 76.25 ± 13.22 1.91±0.68 83.05 ± 31.88 4.49 ± 2.47 55.56 ± 24.28 91.33 ± 6.73 112.71 ± 10.34 <0.0001
6 F (n=10) 2.34±0.61 76.25 ± 9.02 2.17±0.49 84.76 ± 13.76 6.01 ± 2.21 76.17 ± 27.75 93.93 ± 6.86 112.35 ± 8.99 <0.0001
7 G (n=7) 2.08±0.50 69.33 ± 17.65 1.99±0.51 78.03 ± 21.39 5.10 ± 1.73 71.83 ± 30.50 95.63 ± 5.88 112.29 ± 8.24 <0.0001
8 H (n=10) 2.42±0.56 77.81 ± 14.92 2.23±0.44 88.49 ± 14.61 5.74 ± 1.54 70.25 ± 14.05 92.94 ± 5.68 114.26 ± 8.52 <0.0001
9 I (n=15) 2.81±0.56 92.13 ± 8.37 2.68±0.51 107.2 ± 9.12 6.49 ± 1.59 81.12 ± 13.10 95.09 ± 9.85 115.83 ± 11.15 <0.0001

Overall difference is based on two-way ANOVA. Test was performed for comparison of individual pulmonary function test of IWO, C&W, RPF,
Operating Deptt., Coolie, Up-Downers, Residential, Shopkeeper against Control group, where p values in ANOVA are significant p<0.05 by multiple
comparison tests. A = IWO, B = C&W, C = RPF, D = Operating Deptt., E = Coolie, F = Up-Downers, G = Residential, H = Shopkeeper, I = Control

effects. Stable asthma in less than 50% is observed in each
group, whereas fatal asthmatic condition is observed in coolie
(32%) and IWO (20%). This is due to the less PEFR values.
Ventillary impairment data show that severe restrictive de-
fects and severe air flow obstruction are high in coolie fol-
lowed by up-down, residential, IWO and shopkeepers. All
the groups except C&W show moderate restrictive defect as
well as moderate air flow obstruction due to the decline in
FVC and FEV1 values. All the groups are subject to risk.
Coolies are on high relative risk due to the workplace expo-

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 RAILWAY TRAFFIC POLLUTION AND HEALTH EFFECTS 

Table 7:Pulmonary status of different exposed and Control groups (12 to 16 yrs exposure).

Sr. T.G. 12 to 16 Years of exposure

No. FVC (L) FEV1 (L) PEFR (L/s) FEV1/FVC (%) p-value
Obs % Obs % Obs % Obs %

1 A (n=12) 2.21 ± 0.80 78.09 ± 23.81 1.82 ±0.69 84.25 ± 21.01 4.74 ± 1.54 62.53 ± 22.33 89.70 ± 13.43 108.30 ± 16.65 <0.0001
2 B (n=8) 2.35 ± 0.73 84.83 ± 17.28 2.06±0.71 94.93 ± 16.86 5.35 ± 2.11 71.04 ± 22.32 91.69 ± 9.54 114.22 ± 10.16 <0.0001
3 C (n=6) 2.23 ± 1.05 88.49 ± 24.66 1.81±0.44 89.16 ± 10.10 4.34 ± 0.85 64.67 ± 5.66 87.07 ± 15.66 108.12 ± 21.25 <0.0001
4 D (n=8) 2.89 ± 0.49 41.40 ± 20.98 2.62±0.42 97.03 ± 20.5 6.22 ± 1.17 74.49 ± 11.0 91.26 ± 8.90 109.26 ± 9.48 <0.0001
5 E (n=10) 2.16 ± 0.89 77.41 ± 21.47 2.24±0.88 93.72 ± 26.50 6.07 ± 1.99 75.68 ± 26.90 90.29 ± 12.73 107.7 ± 17.53 <0.0001
6 F (n=0) - - - - - - - - -
7 G (n=9) 1.80 ± 0.77 74.07 ± 19.78 1.70±0.65 86.73 ± 20.23 4.21 ± 0.91 65.67 ± 13.36 95.58 ± 6.58 118.79 ± 10.42 <0.0001
8 H (n=13) 2.28 ± 0.94 44.35 ± 25.02 1.90±0.64 83.33 ± 18.79 4.49 ± 1.81 59.23 ± 18.38 84.11 ± 20.10 104.74 ± 27.70 <0.0001
9 I (n=13) 2082 ± 0.28 89.80 ± 9.30 2.59±0.39 101 ± 9.10 6.67 ± 1.61 81.94 ± 17.93 93.37 ± 8.78 116.04 ± 0.95 <0.0001

Overall difference is based on two-way ANOVA. Test was performed for comparison of individual pulmonary function test of IWO, C&W, RPF,
Operating Deptt., Coolie, Up-Downers, Residential, Shopkeeper against Control group, where p values in ANOVA are significant p<0.05 by multiple
comparison tests. A = IWO, B = C&W, C = RPF, D = Operating Deptt., E = Coolie, F = Up-Downers, G = Residential, H = Shopkeeper, I = Control

Table 8: Pulmonary status of different exposed and Control groups (17 to21 yrs exposure).

Sr. T.G. 17 to 21 Years of exposure

No. FVC (L) FEV1 (L) PEFR (L/s) FEV1/FVC (%) p-value
Obs % Obs % Obs % Obs %

1 A (n=9) 2.84 ± 1.38 99.77 ± 45.15 2.40±0.52 102.44 ± 23.53 6.29 ± 1.42 77 ± 14.39 87.75 ± 14.50 106.4 ± 19.16 <0.0001
2 B (n=8) 2.38 ± 0.22 84.87 ± 18.48 2.18±0.23 95.62 ± 16.35 5.94 ± 0.97 75 ± 12.76 95.44 ± 5.50 117.25 ± 7.36 <0.0001
3 C (n=19) 2.56 ± 0.87 82.15 ± 20.15 2.30±0.65 90.89 ± 17.77 6.64 ± 1.74 89.52 ± 26.10 91.98 ± 11.31 112.52 ± 13.76 <0.0001
4 D (n=5) 2.54 ± 0.27 77.6 ± 12.09 2.33±0.32 87.4 ± 14.63 7.09 ± 2.06 82.8 ± 24.13 92.4 ± 13.24 112.6 ± 15.22 <0.0001
5 E (n=16) 2.41 ± 0.62 81.25 ± 19.43 1.96±0.59 80.12 ± 24.45 5.00 ± 1.55 63.56 ± 20.8 85.06 ± 23.05 101.8 ± 27.98 <0.0001
6 F (n=0) - - - - - - - - -
7 G (n=19) 1.96 ± 0.69 67.15 ± 18.32 1.81±0.62 76.67 ± 18.63 5.34 ± 2.46 84.81 ± 35.59 94.26 ± 9.74 109.78 ± 23.90 <0.0001
8 H (n=5) 2.03 ± 0.72 67.8 ± 17.96 1.74±0.52 73.58 ± 19.09 4.50 ± 0.76 58.58 ± 8.50 81.25 ± 28.90 101.6 ± 36.92 <0.0001
9 I (n=12) 2.67 ± 10.56 88.75 ± 10.29 2.28±0.36 96.08 ± 14.65 7.06 ± 1.43 89.08 ± 11.41 87.09 ± 13.26 109 ± 17.21 <0.0001

Overall difference is based on two-way ANOVA. Test was performed for comparison of individual pulmonary function test of IWO, C&W, RPF,
Operating Deptt., Coolie, Up-Downers, Residential, Shopkeeper against Control group, where p values in ANOVA are significant p<0.05 by multiple
comparison tests. A = IWO, B = C&W, C = RPF, D = Operating Deptt., E = Coolie, F = Up-Downers, G = Residential, H = Shopkeeper, I = Control

Table 9: Pulmonary status of different exposed and Control groups (above 22 yrs of exposure).

Sr. T.G. Above 22 Years of exposure

No. FVC (L) FEV1 (L) PEFR (L/s) FEV1/FVC (%) p-value
Obs % Obs % Obs % Obs %

1 A (n=12) 2.14±0.31 70.91 ± 8.4 2.05±0.35 86.5 ± 14.7 6.19±1.10 76.91 ± 14. 95.89 ± 7.1 113.5 ± 31. <0.0001
2 B (n=16) 2.16±0.23 72.5 ± 9.71 2.17±0.27 88 ± 8.07 5.88±1.54 72.37 ± 17.53 96.48 ± 5.68 120.93 ± 7.03 <0.0001
3 C (n=12) 2.45±0.67 77.25 ± 18.84 2.33±0.56 93.66 ± 19.02 6.14±1.66 75.16 ± 17.53 95.87 ± 5.16 124.75 ± 11.37 <0.0001
4 D (n=16) 2.25±0.65 74.64 ± 17.34 2.11±0.58 88.3 ± 18.32 5.58±1.85 69.81 ± 21.36 94.76 ± 4.97 119.25 ± 7.63 <0.0001
5 E (n=8) 1.72±0.96 57.5 ± 29.46 1.17±0.41 53.75 ± 19.99 4.09±1.69 55.12 ± 23.87 88.11 ± 27.66 112.87 ± 35.84 <0.0001
6 F (n=0) - - - - - - - - -
7 G (n=6) 1.52±0.40 63.33 ± 19.89 1.49±0.37 80.16 ± 21.12 4.68±1.34 72 ± 17.58 98.58 ± 1.67 127.5 ± 7.529 <0.0001
8 H (n=4) 1.52±0.92 65.75 ± 31.98 1.15±0.71 65.45 ± 38.16 2.58±1.45 42.11 ± 22.01 76.73 ± 29.95 98 ± 37.76 <0.0001
9 I (n=9) 2.75±0.19 92.33 ± 7.58 2.29±0.39 98.33 ± 14.94 7.02±1.56 87.88 ± 98.94 83.4 ± 11.43 107 ± 17.33 <0.0001

Overall difference is based on two-way ANOVA. Test was performed for comparison of individual pulmonary function test of IWO, C&W, RPF,
Operating Deptt., Coolie, Up-Downers, Residential, Shopkeeper against Control group, where p values in ANOVA are significant p<0.05 by multiple
comparison tests. A = IWO, B = C&W, C = RPF, D = Operating Deptt., E = Coolie, F = Up-Downers, G = Residential, H = Shopkeeper, I = Control

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Kailas F. Khairnar et al.
sure and unhealthy work conditions. Statistical analysis of
all the groups shows significant relationship between dust
exposure and decline in pulmonary function.
&21&/86,21$1'5(&200(1'$7,216
Unhealthy work conditions were observed in all the groups
during the survey. These conditions affect health of the ex-
posed population. Most of the exposed population was una-
ware about the effects of the dust pollution. Group of coolie
is most vulnerable group to respiratory impairment whereas
group of RPF shows fewer effects. In the control group, FVC,
FEV1 and PEFR were close to the expected values, which
show better lung efficiency in unexposed population. The
study concludes that all exposed population to the railway
transportation is vulnerable to respiratory impairment. It is
recommended that equipments like dust collectors must be
kept on railway platform for collection and separation of
dust particles. Tree plantation must be done on either sides
of track which will restrict dust exposure of shopkeepers as
and residential people. IWO, C&W and coolie workers must
use the mask at workplace area. All work places of the groups
should introduce the dust exhaust system wherever possi-
ble. Group of operating, up-down and RPF must use mask
whenever they are exposed to dust on platform. Campaign
should be organized in context of public awareness about
dust and effects of air pollution.
5()(5(1&(6
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Khairnar, K.F. and Ingle, S. T. 2009. Railway transportation noise: A study
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Krelt, J.W., Gross, K.B., Moore, T.B., Lorenzen, T.J., D. Arcy, J. and
Eschenbacher, W.L. 1989. Ozone induced changes in pulmonary func-
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217-222.
Karakatatsani, A., Andreadaki, S., Katsouyanni, K., Dimitroulis, I.,
Trichopoulos, D., Benetou, V. and Trichopoulou, A. 2003. Pulmonary
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nary disease: A nested case-control study in Athens, Greece. Euro-
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Kappos Andreas, D., Bruckman Peter, Eikmann Thomas, Englert Norbert,
Heinrich Uwe, Hoppe Peter, Koch Eckehard, Krause Georg H.M,
Kreyling Wolfgang, G., Rauchfuss Knut, Rombout Peter, Schulz-
Klemp Verena, Thiel Wolf R. and Wichmann, H. Erich 2004. Health
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Sengupta, S. et al. 1996. Assessment of population exposure and risk zones
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Praveen Tamot, Sadhna Tamot* and Ashu Awasthi
Post Graduate Department of Zoology, Govt. Motilal Vigyan Mahavidyalaya (M.V.M.), Bhopal-462 003, M. P., India
*Department of Zoology, Sadhu Vaswani College, Bairagarh, Bhopal, M. P., India

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,1752'8&7,21
The Upper and Lower lakes, popularly called Bhoj wetland
(Ramsar Site) are ecological landmark on the map of Bhopal
city of Madhya Pradesh, India. The Upper lake, situated on
the south-west side of the city, is lifeline for the citizens of
the city whereas the Lower lake is surrounded by urban ag-
glomeration and gives a panoramic view maintaining cli-
mate of the city and also the dividing line between old and
new city. The twin impoundments have made Bhopal “the
city of lakes”.
Unfortunately, Bhopal has no river system and thus, the
lakes and reservoirs are vital for meeting water demand of
ever-growing population of the city. The population of the
city is more than 17 lakhs. In the past, water from the Upper
lake was adequate to meet the water demand of the city, but
due to increasing population in the past three decades, the
lake water became insufficient, therefore, Kolar water sup-
ply (35 km from city) has been installed to meet growing
water demands. In the present scenario even both these wa-
ter bodies are insufficient to meet the growing water needs.
Finally, Government of Madhya Pradesh is going to bring
Narmada river water (80 km from city) to bridge the gap
between demand and the supply of the drinking water. There-
fore it is necessary to conserve and manage the available
aquatic resources in and around the city for future.
The Lower lake is badly managed and under great envi-
ronmental stress due to multiple anthropogenic activities.
As a result, water quality of the lake is getting deteriorated
day by day and also gradually getting filled up leading to
eutrophication.
With the financial assistance from Japan Bank for Inter-
national Co-operation (JBIC), State Government and Gov-
ernment of India executed a major Project on conservation
and management of Bhoj wetland amounting to Rs. 247
crores. The project is comprised of 21 sub-projects, of which
the main are: (i) desilting and deweeding, (ii) afforestation,
(iii) catchment area treatment, (iv) solid waste treatment, (v)
shifting of Dhobi-ghat, (vi) sewage collection diversion and
treatment, (vii) deepening and widening of spill channel,
(viii) aquaculture, (ix) construction of physical barrier (road)
between habitation and lake, (x) public awareness, (xi) lake
view promenade at lake shore, (xii) water quality monitor-
ing, (xiii) strengthening of earthen dam, and (xiv) installa-
tion of aeration units, etc.
Under this sub-project, three different types of aeration
units have been installed to oxygenate the lake water espe-
cially in hypolimnion zone. This activity has not only im-
proved the water quality but also increased the scenic beauty
and became a tourist attraction and provide better environ-
ment and water quality for survival of aquatic fauna
 Praveen Tamot et al.

especially fish. There is a significant difference between f. Unmanaged aquaculture.

fountains and aerators. The primary function of an aerator is g. Pollution from point and non-point sources.
to add oxygen and induce circulation in the water. The pri- h. Idol immersion
mary function of a fountain is to create an aesthetic effect. i. Algal blooms.
The other major advantages of aeration units are: (i) re- 0$7(5,$/6$1'0(7+2'6
duction of growth of algal blooms by converting soluble
phosphorus into non-soluble form, (ii) increase of fish For impact of different types of aeration units on the lake
growth, (iii) increased living space for fish to increase their limnology, samples were collected near the aeration units,
production, (iv) reduction of organic bottom sediment before 6-10 hours prior to the functioning of units and dur-
(muck), (v) prevention of summer/winter kill of fish, and ing the functioning of aeration units from surface and bot-
(vi) overcoming of taste and odour problems. Due to verti- tom of the lake during year 2009 for comparative study.
cal water movement and mixing of lake water, overall dis- Assessment of different physical, chemical and biologi-
solved oxygen level increases in water column. cal parameters was made by the methods of APHA (1995)
The present study focused on the impact of different types and NEERI (1986).
of aeration units: (i) floating fountain, (ii) ozoniser, and (iii) The aeration units: The main objective of installation of
floating fountains cum ozoniser on the lake water. floating fountain was to oxygenate the lake water through
Morphometric features of Lower lake: Some of the im- aeration. The desirable oxygen level in water is essential for
portant morphometric and other features of the Lower lake biotic life of the lake. The three types of aeration units were
are as below. installed under Bhoj Wetland programme and these floating
1. Period of Lake formation : 1794 by Nawab units can be moved in the radius of 500 meter to cover maxi-
Chhote Khan mum water area of the lake. They are as:
2. Type of the dam : Earthen dam a. Floating fountains: Units are simple type of floating
3. Location: (Longitude) : 77°24’-77°26’E fountains which take water from hypolimnion/
(Latitude) : 23°14’-23°15’N metalimnion with higher quantity and the height of jet
4. Catchment area : 9.6 km2 fountain is about 80 feet approximately (Fig. 1).
5. Water spread area : 1.3 km2 b. Ozoniser: Units convert oxygen into ozone and pump
6. Shoreline length : 6.16 km to deeper zone of the lake. These units have no fountain
7. Maximum length : 1.6 km (Fig. 2).
8. Maximum width : 0.9 km
c. Floating fountain cum ozoniser: Units have both the
9. Maximum depth : 9.4 m
systems, fountain as well as conversion of the oxygen in
10. Discharge of wastewater : 7.3 M.G.D. (Million
to the ozone. Height of the jet fountain is comparatively
Gallons per Day)
lower (Fig. 3).
11. Source of water : Rain water, seepage
from Upper lake and 5(68/76$1'',6&866,21
domestic raw sewage
12. Main use of water : Washing, boating and Lake aeration system was first described by Wand & Foley
aquaculture (1919). Shuler (1972) reported that taste and odour prob-
lems can also be overcome to some extent by legation through
Associated problems of Lower lake: Following are the oxidation of reduced organic and inorganic compounds.
important facts which are responsible for the deterioration Chern & Cheng (1997) worked on oxygen transfer model-
of the water quality as well as degradation of the environ- ling of diffused aeration system. Rusan (1971) emphasized
mental status of the lake: use of ozone application for wastewater treatment. Al-
a. Siltation of the lake due to mixing of untreated domestic Ahmady (2006) analysed oxygen transfer performance on
sewage, garbage, etc. The inflow of approximately 7.3 sub-surface aeration systems. Bhuyar et al. (2009) studied
M.G.D. design characteristics of curved blade aerator with respect
b. Washing activities. to aeration efficiency and overall oxygen transfer coefficient
c. Human intervention in fringe area of the lake. and comparison with CFD modelling.
d. Huge quantity of nutrient input in the lake ecosystem Impact of different aeration units on different physico-
due to death and decay of aquatic macrophytes. chemical conditions during pre-operation and during opera-
e. Removal of oxygenated surface water through waste weir tion on surface and bottom waters is shown in Fig. 4. The
at Pul-Pukhta.

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 IMPACT OF DIFFERENT AERATION UNITS ON LIMNOLOGY OF LOWER LAKE, BHOPAL
Fig. 1: Floating fountain
Fig. 2: Ozoniser
Fig. 3: Floating Fountain cum Ozoniser
lake water was found to be of alkaline nature and its pH var-
ied from 7.2-8.4. Slight reduction in pH value was noticed
during operational period of aeration units. The alkalinity is
the sum total of carbonate and bicarbonate. According to
Spence (1967) if the alkalinity is more than 60 mg/L the
lake is nutrient rich. In the present investigation, the range
of total alkalinity varied from 140-220 mg/L. On the basis
of the alkalinity values, Lower lake can be considered nutri-
ent rich. Significant reduction was noted during operation
period of aeration units.
Most of the time free carbon dioxide was absent on sur-
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

face, which may be due to its utilization in photosynthetic
activities of algal blooms and aquatic macrophytes, whereas
higher values were detected at bottom, which may be due to
high rate of organic decomposition with low or no photo-
synthesis. Aeration units play significant role in reduction
of carbon dioxide especially in hypolimnion region.
Dissolved oxygen in water is of great importance to all
aquatic organisms and indicator of water quality especially
the magnitude of eutrophication. Tamot (1998) reported
lower range of dissolved oxygen in hypolimnion zone of
Lower lake, Bhopal. Eutrophicated water bodies have wide
range of dissolved oxygen in which oxygen is more at sur-
face and depletes fast with the depth. In the present investi-
gation wide range of dissolved oxygen was noted from 3.4
to 14.0 mg/L. Floating fountain cum ozoniser and ozoniser
are most effective aeration units for increasing dissolved
oxygen level in water column. Due to lack of oxygen/anaero-
bic condition especially in cloudy weather, mass fish mor-
tality was reported before installation of these aeration units.
After installation of the aeration units, sufficient oxygen
was recorded in water column, which supports better envi-
ronment and water quality for survival and faster growth of
fish.
Biochemical oxygen demand is a good indicator for pres-
ence of organic pollution. Kaushal & Sharma (2000) stated
that high BOD may cause indirect harm by reducing dis-
solved oxygen inimical to fish life and other beneficial
aquatic organisms. In the present investigation, its range var-
ied from 6.2 to 22.0 mg/L with higher values recorded in
hypolimnion zone. Floating fountain cum ozoniser and ozo-
niser play significant role for reduction in BOD.
Nitrate is an important plant nutrient. It is one of the most
oxidized forms of nitrogen and occur in varying amount in
the lake water depending upon the nature of the water shade,
season, degree of pollution and abundance of planktonic life
(Rodhe 1969, Sommer 1989). Its range varied from 0.3 to
1.6 mg/L. Reduction in nitrate value was noted during op-
eration of aeration units. Bengtssen & Gelin (1975) reported
that aeration can be an effective method to reduce the nutri-
ent concentration of the lake and improve overall water
quality.
Cowell et al. (1987) supported the present findings. He
worked on impact of lake aeration on the limnology of a
hyper-eutrophic lake in central Florida and found signifi-
cant increase in dissolved oxygen at bottom and decline in
pH and alkalinity values, and reduction in the density of blue
green algal blooms.
In Lower lake more than 20 fish species were reported,
most fish were: Ctenopharygodon idella, Catla catla, Labeo
 Praveen Tamot et al.
Fig.4: Impact of aeration units (Floating Fountain: FF, Ozoniser and Floating Fountain cum Ozoniser on different physical and chemical parameters
during pre-operation (P-O) and during operation (D-O) in surface (S) and bottom (B) waters.
rohita, Chana punctatus, Mastacembelus armatus, Mystus
seenghala and Wallago attu. Due to installation of aeration
units, dissolved oxygen level in water column was found
satisfactory and no mass fish mortality recorded.
&21&/86,21
On the basis of the present observations, all three types of
aeration units play a significant role in lake limnology. But,
Floating fountain cum ozoniser has given the best results in
reduction of the BOD, free carbon dioxide and total alkalin-
ity, and increasing dissolved oxygen and nutrient loading.
Ozoniser unit was most effective for controlling the growth
of bacteria. Floating fountain is also effective for increasing
dissolved oxygen level and also the scenic beauty of the lake.
5()(5(1&(6
Al-Ahmady Kossay, K. 2006. Analysis of oxygen transfer performance on
sub-surface aeration systems. Int. J. Environ. Res. Public Health, 3(3):
301-308.
APHA 1995. Standard Methods for the Examination of Water and
Wastewater. 6th Edition, American Public Health Association, Wash-
ington DC.
Bengtssen, L. and Gelin, C. 1975. Artificial aeration and suction dredging
method for controlling water quality. Proc. of Symposium, 12.
Bhuyar, L.B., Thakre1 S.B. and Ingole, N.W. 2009. Design characteristics
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
of curved blade aerator w.r.t. aeration efficiency and overall oxygen
transfer coefficient and comparison with CFD modeling. Int. J. Engi-
neering, Science and Technology, 1(1): 1-15.
Chern Jia-Ming and Cheng-Fu Yu 1997. Oxygen transfer modeling of dif-
fused aeration system. Ind. Eng. Chem. Res., 36(12): 5447-5453.
Cowell, B.C., Daves, C.J., Gardiner William, E. and Scheda, M. Sandra
1987. The influence of whole lake aeration on the limnology of a
hypereutrophic lake in central Florida. Hydrobilogia, 148: 3-24.
Kaushal, D.K. and Sharma, V.K. 2000. Limnology and productivity of
Badkhal lake, Haryana. Ab. Indian Fish Sc. Cong., pp. 38.
NEERI 1986. Manual of Water and Wastewater Analysis. National Envi-
ronmental and Engineering Research Institute, Nagpur.
Rodhe, W. 1969. Crystallization of eutrophication concept in North Eu-
rope. In: Eutrophication, Causes, Consequence and Correction, Nat.
Acad. Nat. Res. Council, Pub.1700: 50-64.
Rusan, H.M. 1971. Ozone generation and its relationship to the economi-
cal application of ozone in wastewater treatment. Baltimore Maryland.
Shuler, J. 1972. How a lake aeration system overcome water consumer
complaints? Public Work 103(5): 92-93.
Sommer, V. 1989. Nutrient status and nutrient competition of phytoplankton
in a shallow hypereutrophic lake. Limnol. Oceanogr., 34: 1162-1173.
Spence, D.H.N. 1967. Factors controlling the distribution of freshwater
macrophytes with particular reference to the lochs of Scotland. J. Ecol.,
55: 147-170.
Tamot Praveen 1998. Water quality monitoring of Lower lake, Bhopal.
Technical Report, pp. 33-34.
Wand Scott and Foley, A.L. 1919. A method of direct aeration of stored
waters. Proc. Ind. Acad. Sci., pp. 71-73.
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Lakshman Nayak and Pragnya Padhi
P. G. Department of Marine Sciences, Berhampur University, Berhampur-760 007, Orissa, India

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,1752'8&7,21 ers erstwhile Kanika and Kujang Zamindary area. It includes

Bhitarkanika is endowed with a very complex and dynamic
ecosystem and is highly fragile in nature. The ecosystem is
complex in a sense that all the sub ecosystems namely fresh-
water, marine and terrestrial are intricately mixed with each
other. The essential factor for maintenance of such ecosys-
tem is regular influx of freshwater from adjoining land and
tidal inflow from the sea. Any change in the regime of either
factor is likely to affect a corresponding change in the man-
grove ecosystem. The Bhitarkanika mangrove forest is the
second largest mangrove forest in India. The mangrove for-
ests all along the Orissa coast are threatened due to high den-
sity of population in these areas and demand for land for
agriculture and shrimp farming. The mangrove belt in
Kendrapada district is called the Bhitarkanika mangrove for-
est, comprising areas between Dhamara mouth and Barunei
coast. It has been notified as Bhitarkanika Wildlife Sanctu-
ary in April 1975 covering an area of 672 sq. km. The core
area of sanctuary has been declared as Bhitarkanika National
Park in September 1998 covering an area of 145 sq. km. The
Bhitarkanika area has been designated as the second Ramsar
site (i.e., Wetland of International Importance) of the State
in August 2002. It is a unique area with rich biodiversity as
it covers different ecosystems such as landmass, tidal water
bodies of the deltaic region, estuaries and territorial waters
of the Bay of Bengal along with their associated flora and
fauna. The proposed Bhitarkanika Biosphere Reserve cov-
at present three protected areas namely: Bhitarkanika Wild-
life Sanctuary, Bhitarkanika National Park and the
Gahirmatha Wildlife Sanctuary. The area supports rich
biodiversity including mangroves and mangroves associated
largest population of estuarine crocodiles and the rare white
crocodiles etc. The mangrove ecosystem plays a vital role in
contributing to the food web, in general, and detritus food
chain in particular, supporting rich estuarine and adjacent
marine fisheries (Chadha & Kar 1999). The mangroves play
a significant role in protecting the internal against cyclones
and the ingress of seawater during tidal surge. Mangroves
stabilize coastal landmass against sea erosion. Mangrove
forests act as a buffer against any natural disasters (Nayak et
al. 2009).
Several works have been done on distribution of man-
groves in India (Sidhu 1963, Nayak 2006). Some work has
been done on the floral characteristics of Bhitarkanika by
Banerjee & Das (1972), Mishra & Panigrahy (1978), Pattanaik
& Choudhury (1989) and Banarjee & Rao (1990). Few works
have been carried out on the hydrological features of Mahanadi
estuary by Upadhyay (1998) and Raju et al. (2000). No work
has been done on saltwater crocodiles in relation to physico-
chemical characteristics from Bhitarkanika Sanctuary. There-
fore the present study is an attempt to study the conservation
and management of salt water crocodile (Crocodylus porosus)
in relation to some important physico chemical parameters
 Lakshman Nayak and Pragnya Padhi
of Bhitarkanika Sanctuary, Orissa.
0$7(5,$/6$1'0(7+2'6
The saltwater crocodiles are endangered species belonging
to the Order Crocodylia and Family Crocodylidae. The com-
mon English name for this crocodile is salt water crocodile
or estuarine crocodile and the scientific name is Crocodylus
porosus. The study was carried out during January 2008 to
December 2008. Bhitarkanika is situated between 20° 04’
and 20° 08’ N latitude and 86°45’ and 87°05’ E longitude.
The monthly water samples were collected from three dif-
ferent stations i.e., 1st is Nalitapatia, 2nd is Dangmal located
between Nalitapatia and Bhitarkanika estuary, and the 3rd
station is chosen at Bhitarkanika estuary, which is towards
the sea where the physico-chemical parameters are highly
fluctuative. The hydrological parameters like temperature,
pH, salinity and dissolved oxygen were studied at monthly
interval of time. The air and water temperatures were meas-
ured using a centigrade thermometer of ± 0.1°C accuracy.
The water samples were brought to the laboratory. The pH
was measured using a digital pH meter. The samples were
analysed for salinity and dissolved oxygen following the
standard methods (APHA 2005). The standard deviation for
each sample was calculated. The secondary data on salt wa-
ter crocodile population were collected and the percentage
of increasing and decreasing of salt water crocodile popula-
tion was calculated for the years 1995-96 to 2009-10.
5(68/76
The hydrological data collected from the three stations of
Bhitarkanika are represented in Tables 1, 2 and 3. The popu-
lation of the crocodile (Crocodylus porosus) in Bhitarkanika
during the year 1995-96 to 2008-09 is given in Table 4. In
the Station 1, the air temperature was highest being
33.1±1.150°C during May, and lowest being 27.3±1.044°C
during January (Table 1). The water temperature was high-
est being 33.0±1.148°C during May, and lowest being 27.6
±1.148°C during December. The pH value was highest
(8.8±0.593) in the month of December, and the lowest
(6.8±0.521) in the month of October. The salinity was high-
est of 20.9±0.914 ppt in the month of April, and the lowest
of 8.1±0.569 ppt in August. The dissolved oxygen was high-
est at 8.5±0.583 mL/L during April, and the lowest at
5.4±0.464 mL/L during November at Station 1.
At Station 2, the air and water temperatures were highest
(34.5±1.174°C, 36.1±1.201°C) during May, and lowest
(27.5±1.048°C, 28.2±1.1.062°C) during January respec-
tively. The pH value was highest being 8.4±0.579, and the
lowest being 7.0±0.529 in December and August respec-
tively. The salinity was highest with a values of 27.1±1.041
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
ppt in April, and lowest of 13.0±0.721 ppt in July. The dis-
solved oxygen concentration was highest being 9.9±0.629
mL/L, and lowest being 5.1±0.451 mL/L during July and
February respectively.
Similarly at Station 3, the highest air temperature was
35.0±1.183°C, and lowest 28.7±5.357°C during the months
of May and January respectively. The water temperature was
highest being 35.1±1.184°C in May, and the lowest being
28.2±1.062°C during January. The pH value was highest at
9.2±0.606 in December, and lowest at 6.7±0.517 in May.
The salinity was highest (31.2±1.117 ppt) in May, and low-
est (12.8±0.715 ppt) August. The dissolved oxygen was high-
est with 9.9±0.629 mL/L in July, and the lowest with
5.2±0.456 mL/L in January. The saltwater crocodile is per-
haps the largest living reptile in the World. During 1975, H.
R. Bustard has coordinated a Project by Government of In-
dia/FAO/UNDP on crocodile breeding and management.
Bustard captive breeding of saltwater crocodiles was initi-
ated by the Forest Department of Government of Orissa in
1975 at Dangamal, the Bhitarkanika Wildlife Sanctuary
(BWLS). The estimated population of crocodiles during 1976
at Bhitarkanika was 96 (Gopi & Pandav 2009). The saltwa-
ter crocodile (Crocodylus porosus) population of 14 years
is represented in Table 4. The highest population of saltwa-
ter crocodile was observed to be 1610 during the year 2009-
10, and the lowest of 511 during the year 1995-96. The high-
est increased percentage of saltwater crocodile population
was observed to be +26.58 % during the year 1999-2000,
and the lowest decreased population of -01.68 % during the
year 2002-03.
Salt water crocodile population varied from station to
station in Bhitarakanika (Table 5). The highest population
was observed to be 580 at Station 3, and lowest of 398 at
Station 1. Crocodile population showed a perfect positive
correlation with air temperature, salinity and dissolved oxy-
gen (r = 0.999, 0.997, 0.946) respectively. Likewise croco-
dile population showed a positive correlation with water tem-
perature and pH, being r = 0.795 and 0.323 respectively (Ta-
ble 6).
',6&866,21
The air temperature varied from 27.3 to 35°C during the study
period. There has been a gradual decrease in temperature from
Station 1 to Station 3 towards the downstream along the river.
The findings are in agreement with the results of Upadhyay
(1988) and Raju et al. (2000). The water temperature varied
from 27.6 to 36.1°C during the study period. The tempera-
ture is slightly higher than the results of Nayak et al. (2009)
from the same environment. This may be due to collection
of samples in different times of the day and other environ-
 MANAGMENT OF SALT WATER CROCODILE IN BHITARKANIKA SANCTUARY 

Table 1: Monthly average values of physico-chemical parameters of Bhitarkanika during January 2008 to December 2008 at Station 1.

Months/Parameter Air Temperature Water Temperature pH Salinity Dissolved Oxygen

(°C) (°C) (ppt) (mL/L)

January 2008 27.3± 1.044 29.1±1.078 8.2±0.572 18.8±0.867 6.6±0513

February 2008 28.0±1.058 29.8±1.091 8.7±0.589 18.9±0.869 8.1±0.569
March 2008 30.3±1.100 29.1±1.078 8.6±0.586 19.1±0.874 7.0±0.529
April 2008 31.3±1.118 31.9±1.129 8.2±0.572 20.9±0.914 8.5±0.583
May 2008 33.3±1.150 33.0±1.148 7.8±0.558 17.8±4.219 7.8±0.558
June 2008 29.1±1.078 32.5±1.140 7.7±0.554 9.0±0.6 8.4±0.579
July 2008 28.1±1.060 31.9±1.129 7.2±0.536 8.3±0.576 7.0±0.529
August 2008 29.7±1.089 29.1±1.078 7.9±0.562 8.1±0.569 6.1±0.493
September 2008 28.3±1.063 30.9±1.111 7.4±0.544 9.2±0.606 6.2±0.497
October 2008 30.0±1.095 32.1±1.133 6.8±0.521 11.0±0.663 5.5±0.469
November 2008 29.8±1.091 31.3±1.118 8.3±0.576 11.9±0.689 5.4±0.464
December 2008 30.5±1.104 27.6±1.048 8.8±0.593 12.0±0.692 6.8±0.521

Table 2: Monthly average values of physico-chemical parameters of Bhitarkanika during January 2008 to December 2008 at Station 2.

Months/Parameter Air Temperature Water Temperature pH Salinity Dissolved Oxygen

(°C) (°C) (ppt) (mL/L)

January 2008 27.5±1.048 28.2±1.062 8.1±0.569 24.2±0.983 6.0±0.489

February 2008 30.2±1.099 29.7±1.089 8.2±0.572 23.8±0.975 5.1±0.451
March 2008 31.0±1.113 31.7±1.126 7.3±0.540 22.4±0.946 7.3±0.540
April 2008 34.4±1.173 34.1±1.167 8.0±0.565 27.1±1.041 6.9±0.525
May 2008 34.5±1.174 36.1±1.201 8.2±0.572 16.3±0.807 8.0±0.565
June 2008 33.9±1.164 34.8±1.179 7.5±0.547 14.2±0.753 7.2±0.536
July 2008 33.6±1.159 34.1±1.167 7.6±0.551 13.0±0.721 9.9±0.629
August 2008 33.1±1.150 33.1±1.150 7.0±0.529 16.0.±0.800 9.6±0.619
September 2008 32.1±1.133 31.9±1.129 7.2±0.536 17.1±0.827 8.8±0.593
October2008 32.0±1.130 30.8±1.113 7.9±0.562 20.2±0.898 8.4±0.579
November 2008 31.4±1.120 32.0±1.131 8.1±0.569 20.1±0.896 8.3±0.576
December 2008 28.1±1.060 29.3±1.082 8.4±0.579 19.1±0.874 7.1±0.532

mental conditions. The water temperature was slightly higher
as compared to the air temperature during winter season,
which is due to absorption of heat by the surface layer (Nayak
1991). The value of pH was quite low during June because
of the organic matter carried out by the flood water into the
riverine system (Raju et al. 2000). The Salinity values ranged
from 8.1 to 31.3 ppt during the study period. Nayak et al.
(2009) have studied the salinity from Bhitarkanika which
ranged from 9.0 to 33.8 ppt coinciding with the present re-
sult. The salinity values increased both in post-monsoon and
pre-monsoon months. The high salinity values may be due
to high evaporation and low intensity of river runoff into
the estuarine system during this period. The dissolved oxy-
gen value showed wide range of variation throughout the
year. The value varied from 5.1 to 9.9 mL/L. The value ranged
from 2.325 to 7.250 mL/L as studied by Nayak et al. (2009),
which is coinciding with the present result. The low value
of dissolved oxygen may be due to biodegradation of or-
ganic matter in water (Raju et al. 2000).
Environment in relation to crocodiles: The saltwater croco-
dile (Crocodiles porosus) is a carnivorous and scavenger
species. It plays a vital ecological role as master predator in
the aquatic habitat where it lives. It feeds on weak and dis-
eased fish and animals. It maintains genetic quality, by its
habit of selective feeding, which controls predatory fish
population. Thus, presence of crocodile actually helps to
increase yield of edible fish for man. Crocodiles are com-
mercially and economically important because their skin has
a high demand in the national and international market for
making shoes and bags which are long lasting and beautiful.
Crocodiles are long lived and a female crocodile produces
about 30 eggs per year. Fifty percent of its eggs can reach up
to culling size within 3-4 years in captivity. These 15 croco-
diles will have a gross value of Rs. 1,56,000. A female croco-
dile can remain productive for about 30 years, which will
worth about Rs. 4,68,000. The crocodiles, in general, keep
the environment clean and maintain the population of un-
wanted fishes in the environment. Therefore, it is our ob-
ligatory duty to protect and conserve the rare and endan-
gered species like saltwater crocodile in Bhitarkanika
sanctuary.

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Lakshman Nayak and Pragnya Padhi

Table 3: Monthly average values of physico-chemical parameters of Bhitarkanika during January 2008 to December 2008 at Station 3.

Months/Parameter Air Temperature Water Temperature pH Salinity Dissolved Oxygen

(°C) (°C) (ppt) (mL/L)

January 2008 28.7±5.357 28.2±1.062 7.9±0.562 24.7±0.993 5.2±0.456

February 2008 30.6±1.106 30.8±1.109 7.6±0.551 24.2±0.983 7.1±0.532
March 2008 32.9±1.147 32.3±1.136 7.8±0.558 21.3±0.923 7.0±0.440
April 2008 34.5±1.174 34.1±1.167 7.1±0.532 31.1±1.115 6.3±0.501
May 2008 35.0±1.183 35.1±1.184 6.7±0.517 31.2±1.117 6.9±0.525
June 2008 34.4±1.173 34.0±1.166 7.4±0.544 15.8±0.794 7.9±0.562
July 2008 34.0±1.166 33.3±1.154 7.3±0.540 13.8±0.742 9.9±0.629
August 2008 32.3±1.136 32.0±1.131 7.2±0.536 12.8±0.715 9.4±0.613
September 2008 32.1±1.133 31.3±1.118 7.1±0.532 18.7±0.864 8.2±0.572
October 2008 31.9±1.129 32.2±1.117 7.0±0.529 21.0±0.916 5.6±0.473
November 2008 31.1±1.115 30.3±1.100 7.2±0.536 20.1±0.896 6.0±0.489
December 2008 29.8±1.091 30.0±1.095 9.2±0.606 22.8±0.954 7.3±0.540

Table 4: The number of saltwater crocodile (Crocodylus porosus) in Bhitarkanika from the year 1995-96 to 2009-10.

Year Hatchling Yearling Juveniles Sub-Adult Adult Total % of Increase or

Decrease

1995-96 304 71 34 92 10 511 -

1996-97 136 232 161 63 68 660 +22.57
1997-98 252 106 121 113 76 668 +1.19
1998-99 149 146 160 144 72 671 +0.44
1999-00 319 181 123 145 146 914 +26.58
2000-01 341 277 237 136 107 1098 +16.75
2001-02 441 340 187 145 217 1330 +17.44
2002-03 438 333 184 140 213 1308 -01.68
2003-04 525 303 210 100 220 1358 +3.68
2004-05 402 279 458 106 204 1449 +6.28
2005-06 657 283 196 121 197 1454 +0.34
2006-07 503 466 257 132 224 1482 +1.88
2007-08 538 342 227 139 252 1498 +1.06
2008-09 529 374 256 144 267 1572 +4.70
2009-10 519 373 298 150 270 1610 +2.36

Source: Forest Department, Government of Orissa, Dharitri, 15th January, 2010.

Table 5: Availability of saltwater crocodile (Crocodylus porosus) in relation to physico-chemical parameters from different stations.

Stations Crocodile Air Temperature Water Temperature pH Salinity Dissolved

Population (°C) (°C) (ppt) Oxygen(mL/L)

1. Nalitapatia 398 33.3 33 8.8 20.9 8.5

2. Dangmala 520 34.5 36.1 8.4 27.1 9.9
3. Bhitarakanika Estuary 580 35 35.1 9.2 31.2 9.9

Table 6: Correlation of saltwater Crocodile (Crocodylus porosus) population with different physico-chemical parameters.

Crocodile Air Temperature Water Temperature pH Salinity Dissolved

Population (°C) (°C) (ppt) Oxygen(mL/L)

Crocodile population 1
Air temperature 0.999232 1
Water temperature 0.795523 0.818657 1
pH 0.323473 0.286143 -0.31602 1
Salinity 0.997035 0.993253 0.746537 0.395329 1
Dissolved oxygen 0.946237 0.958187 0.948753 -1.3E-15 0.91854 1

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 MANAGMENT OF SALT WATER CROCODILE IN BHITARKANIKA SANCTUARY
0$1$*(0(17 675$7(*,(6
The broad strategy adopted for rehabilitation of crocodiles
was to protect them in their natural habitat to enhance the
population quickly through captive breeding (rear and re-
lease) and to build up trained personnel for the job. The broad
objectives and activities under the crocodile project are the
following:
1. To protect the remaining population of crocodilians in
their natural habitat by creating sanctuaries.
2. To rebuild natural population quickly through grow and
release or rear and release technique involving the fol-
lowing phases of operation:
• Collection of eggs from natural nests as soon as these
are laid.
• Incubation of these eggs under ideal temperature and
humidity maintained in artificial hatcheries.
• Hatchlings are reared up to the young crocodiles in ideal
captive-husbandry conditions.
• Marking and releasing young crocodiles in protected area
and assessing the result of release along with protection
of the released crocodiles.
3. To promote captive breeding.
4. To take up research work on behavioural biology includ-
ing reproduction, thermo-regulation, feeding, water ori-
entation, locomotion, etc. in order to improve the man-
agement plan.
5. To build up a level of trained personnel for better conti-
nuity of the project through training imparted at the
project sites and through the Central Crocodile Breed-
ing and Management Training Institute, Hyderabad.
6. To involve the local people intimately in the crocodile
breeding project. They should be engaged in collection
of eggs and rearing of the crocodiles. The other local fish-
ermen population should be provided with alternative
source of income during no fishing season i.e., during
the breeding and nesting periods of the crocodiles, so
that they will not harm to the crocodile.
Conservation of crocodiles: Crocodilians were threatened
in India due to indiscriminate killing for commercial pur-
poses and severe habitat loss until enforcement of the Wild-
life (Protection) Act, 1972.
All three species of crocodiles (Gharial, Mugger and
Saltwater crocodile) in the river systems of Orissa were on
the verge of extinction during 1970s. So the conservation of
crocodiles was felt most urgent. The following measures
should be taken for conservation of crocodiles.
• Crocodiles were very few because of ever-increasing hu-
man activity in the rivers and their other traditional habi-
tats, and consequent reduction in the extent of habitable
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

stretches. So crocodile habitat protection areas should
be identified.
• The survival rate of the crocodile hatchings in the nature
is low because of predation. Therefore, captive breeding
is required for better survival rate of the crocodiles.
• The Gharial and Saltwater Crocodile Conservation Pro-
gramme was first implemented in Orissa in early 1975
and subsequently the Mugger Conservation Programme
was initiated, since Orissa is having distinction of exist-
ence of all the three species of Indian crocodilians. The
funds and technical support for the project came from
UNDP/ FAO through the Government of India should
be exerted to other areas and for longer period.
• Public-relation should be among the local people to make
aware of the importance of the crocodiles in ecosystems.
• Habitat development inside the sanctuary should be done
with funds received from MoEF, Govt. of India for plan-
tations, digging and renovation of creeks and digging of
ponds.
• To wean the poachers away from poaching, a massive
awareness programme should be undertaken for anti-
poaching strategies.
• Encroachment on the mangrove habitat for rehabilita-
tion, agriculture and prawn culture, etc. should be banned.
• Use of gill nets in the rivers, creeks and estuaries through-
out the sanctuary should be strictly prohibited.
• Degraded mangrove forest should be taken up for resto-
ration, plantation and rigid protection. Moreover, at least
100 meter wide strip of mangrove forest should be cre-
ated all along the rivers/creeks adjacent to cultivated land
inside the sanctuary.
• Measures should be taken to eliminate man-crocodile
conflict.
• Studies on the ecology of estuary/saltwater crocodiles
should be continued considering the existing gap in the
knowledge regarding various aspects of ecology of the
species in Bhitarkanika and other distributional range in
Orissa as well as in the entire country.
• Education and awareness should be created among the
local inhabitants to change their hostile attitude towards
conservation of saltwater crocodiles and other wildlife
species along with the threatened mangrove ecosystem.
&21&/86,21
The endangered species crocodiles are threatened in India
due to indiscriminate killing for commercial purposes and
severe habitat loss. The saltwater crocodile (Crocodylus
porosus) in the river systems of Orissa was on the verge of
extinction during 1970s and still now its population is very
low. Crocodiles were very few because of ever-increasing
human activity in the rivers and their other traditional
 Lakshman Nayak and Pragnya Padhi
habitats and consequent reduction in the extent of habitable
stretches. Despite best efforts of rearing and rehabilitant, for
this species, the future can not be bright unless the sanctuary
and the crocodiles are adequately protected and large number
of juvenile saltwater crocodiles attain breeding size and com-
mence breeding in the wild. A long term research programme
on crocodile habitat is highly essential at Central Institutes,
Universities and NGOs in order to protect and conserve the
saltwater crocodiles.
5()(5(1&(6
APHA 2005. Standard Methods for the Examination of Water and
Wastewater. American Public Health Association, AWWA and WPCF,
Washington DC.
Banerjee, L.K. and Das, G.C. 1972. New distributional records from Orissa
coast. Bulletin of Botanical Survey of India, 14(1-4): 184-186.
Banerjee, L.K. and Rao, T.A. 1990. Mangroves of Orissa Coast and Their
Ecology. Bishen Singh Mahendra Pal Singh Publisher, Dehradun, pp.
1-114.
Chadha, S. and Kar, C.S. 1999. Bhitarkanika. Myth and Reality. Natraj
Publisher, Dehradun, pp. 55-170.
Dharitri, 15th January, 2010. Bhitarakanika re Baula Kumbhira Sankhya
1610. pp. 1-2.
Gopi, G.V. and Pandav, B. 2009. Human sharing space with Crocodylus
porosus in Bhitarkanika Wildlife Sanctuary: conflicts and options.
Current Sciences. 96(4):459-460.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
Mishra, S.C. and Panigrahi, G. 1978. Studies on mangrove flora of Orissa
with particular reference to the Rhizophoraceae. RBR Journal of Eco-
nomic and Taxonomic Botany, 11(1): 121-132.
Mohanty, S.C., Kar, C.S., Kar, S.K. and Singh. L.A.K. 2004. Wild Orissa,
Wild life Organization, Forest Department, Govt. of Orissa,
Bhubaneswar, pp. 19-73.
Nayak, L. 1992. Studies on Seasonal Distribution and Abundance of the
Post Larvae of Penaeid Prawns in Relation to Biological and Physico-
chemical Factors at Rushikulya Estuary. Ph.D. Thesis, Berhampur
University, pp. 1-209.
Nayak, L. 2006. Mangrove forests of Orissa. Journal of Indian Ocean Stud-
ies, 14(3): 450-458.
Nayak, L., Behera, D., Mohapatra, R. and Swain, D. 2009. A study on
Bhitarkanika forest: A sensitive fragile ecosystem. Nature Environ-
ment and Pollution Technology, 8(1): 43-47.
Pattanaik, S.N. and Choudhury, B.P. 1989. Present status and future devel-
opment of mangrove vegetation in Bhitarkanika. In: Indo-US Work-
shop on Wetlands, Mangroves and Biosphere Reserve, pp. 89-95. Gov-
ernment of India, Ministry of Environments and Forests, New Delhi.
Raju, V.A., Nayak, L. and Das, R.C. 2000. Seasonal abundance of post
larvae of paneid prawns in the Mahanadi estuary, Orissa. Asian J. Zool.
Science, 9: 15-23.
Shriadah, A.M. 2000. Chemistry of the mangrove waters and sediments
along the Arabian Gulf shoreline of the United Arab Emirates. Indian
Journal of Marine Sciences, 29: 224-229.
Sidhu, S.S. 1963. Studies on Mangroves of India. Proceedings of Indian
Academy of Sciences, 33(8): 129-136.
Upadhyay, S. 1998. Physico-chemical characteristics of the Mahanadi es-
tuarine ecosystem, east coast of India. Indian Journal of Marine Sci-
ences, 17: 19-23.
 0CVWTG'PXKTQPOGPVCPF2QNNWVKQP6GEJPQNQI[ ,661 9RO 1R SS 
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J. Jean Jose, P. Udayakumar, M. P. Deepak, B. R. Rajesh, K. Narendra Babu and A. Chandran*
Chemistry and Marine Biology Laboratory, Chemical Sciences Division, Centre for Earth Science Studies,
Thiruvananthapuram-635 031, Kerala, India
*School of Applied Life Sciences, Mahatma Gandhi University Regional Centre, Pathanamthitta-689 645, Kerala, India

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,1752'8&7,21 the south-west coast of India very little is known of

Marine pollution management is based on monitoring vari-
ous physico-chemical and biological parameters to detect
changes in the environment. Recent studies carried out in
the coastal waters of India reveal that the coastal belt includ-
ing the water and sediment is continuously being threatened
by various pollutants discharged directly from the industrial
and effluent activities (Govindasamy & Azariah 1999). Com-
pared with the east coast, the Bay of Bengal, the west coast
of India, the Arabian Sea, is highly productive due to the
seasonal upwelling processes in association with the south-
west monsoon and river inflow (Madhupratap et al. 2001,
Venkataraman & Wafar 2005). Some of the recent studies
carried out in the south-west coast of India inferred the main
source of pollution inflow from anthropogenic activities and
industrial origin, threatening the coastal biodiversity to a risk
(Ouseph et al. 2009, Udayakumar et al. 2009).
The importance of zoobenthic communities in pollution
studies is well established. It is believed that the sea-bed acts
as a sink for most of the pollutants entering the marine eco-
system, and the more stable sediments along with their in-
habitant fauna give a clear picture of the severity of con-
taminants and act as an indicator species of that particular
area (Gray 1981). According to Gesteira & Dauvin (2005)
marine benthic organisms exhibit moderately fast response
to stress, and being predominantly sessile or slow moving
organisms, are vulnerable to the effects of sediment contami-
nation, assimilating the effects of pollutants over time. In
polychaetes as indicator species of coastal pollution. The
available information, however, deals mainly with their abun-
dance in mangrove swamps, as a demersal fishery indicator,
their species succession in soft bottom sediments, their com-
munity structure in offshore water sand as an anthropogenic
indicator (Sunil Kumar & Antony 1994, Sunil Kumar 2000,
Harkantra & Rodrigues 2003, Thomas et al. 2006, Joydas et
al. 2009, Soniya Sukumaran & Sarala Devi 2009). The ob-
jective of the present work was to study the seasonal abun-
dance of meio, micro and macro level polychaete genus
Glycera in relation to textural characteristics of the sediments,
and to confirm its role as an organic enrichment indicator
along the south-west coast of India.
0$7(5,$/6$1'0(7+2'6
Study sites: The sampling locations (Fig. 1) taken for the
present study were Neendakara (8°57’29”N and 76°26’20”E),
Alleppey (9°29’28”N and 76°13’33”E), Cochin (9°56’18”N
and 76°09’13”E), Ponnani (10°47’06”N and 75°49’49”E) and
Kasargod (12°28’56”N and 74°53’19”E). All the sampling
points have a distance of 10 km from shore. These stations
are all biologically productive and considered as the major
fishing zones of Kerala. The coastal waters of these stations
were subjected to discharge from industrial and anthropo-
genic activities, and pollutants arising from fish landing and
seafood processing centres.
Sampling and laboratory procedures: The sampling was
 J. Jean Jose et al.

carried out seasonally during the year 2008. Four replicate genus habitat preference and abundance. From this it is in-
samples for each sediment samples from the five stations ferred that meio type Glycera shows great affinity to sedi-
were collected using Van-Veen grab of 0.04 m2 operated ment texture and organic carbon throughout the study. The
onboard Coastal Research Vessels (CRV) Sagar Purvi and meio type Glycera polychaete was represented more in most
Sagar Paschimi. The collected sediment samples were sorted of the stations, especially Neendakara, Cochin, Ponnani and
into two sets. Those for chemical analysis, especially for tex- Kasargod.
ture and organic matter content, were sealed in plastic bags
and frozen. For zoo benthos collection, the sediments were ',6&866,21$1'&21&/86,21
separated through three sieves (mesh size of the upper, mid- Polychaetes are one of the most characteristic groups of soft-
dle and lower layer sieves 0.5 mm, 0.1 mm and 60 µ respec- bottom benthic organisms and are an important component
tively) of diameter 20.5 cm placed layer by layer in a plastic of benthic communities (Arvanitidis et al. 2002). In the
tray using continuous flow of seawater by means of the fa- present study, the polychaetes, especially the meio category
cility provided in the deck of the research vessel. The col- of the genus Glycera, were found as the major module among
lections were preserved in 1.0 % formalin containing Rose them. The results of the present observations revealed that
Bengal stain. The zoobenthic organisms entangled in the the genus Glycera occurs in areas where the percentage of
upper, middle and lower sieves were grouped based on their silt and clay components of sediments were more than that
length as micro (100 to 500 µ), macro (> 500 µ) and meio of the sand fraction having rich organic carbon content. The
category (50 to 100µ). The sorting of meio and micro sand dwelling habit of polychaetes along the south-west coast
polychaetes from fine sediments was carried out using of India, especially in the Cochin mangrove ecosystem, is
decantation techniques as described by Mcintyre & Warwick well documented (Sunil Kumar & Antony 1994).
(1975). Polychaete counts were expressed in numbers/m2.
The genus level identification of polychaetes was made ac-
cording to Fauvel (1953) and Fauchald (1977). The sedi-
ment texture (sand, silt and clay) analysis was determined
by pipette analysis (Krumbein & Pettijohn 1938) and or-
ganic carbon of the composite samples using wet digestion
(chromic acid) followed by back titration with ferrous am-
monium sulphate (EL Walkeel & Riley 1957).
A dendrogram for hierarchical classification was used to
assess the potential relationship between the polychaete gen-
era, sediment texture and organic carbon content. Cluster
analysis was performed using Ward’s hierarchical agglom-
erative method and Euclidean distance measure. Statistical
analysis was carried out using Systat 8.0 software.
5(68/76
The abundance of the polychaete genus Glycera, commonly
known as the marine blood worm was found dominant in all
the sampling stations. The other opportunistic invader
polychaete genera in the study areas were Ancistrosyllis,
Heteromastides, Nephthys, Neries, Paraheteromastus,
Polydora, Sabellidae and Syllis. Cluster analysis was applied
to find out the similar genera of polychaetes between the
sampling stations having sediment texture rich in sand silt,
clay and organic carbon. It resulted in a dendrogram (Fig. 2)
in to six statistically meaningful clusters. Cluster 1 showed
the abundance of micro type Glycera and macrotype
Polydora and Neries in the sediment texture rich in sand.
The communities interrelationship of different polychaete
genera at the sampling stations was more pronounced in the
Fig. 1: Map of Kerala showing the sampling points.
clusters 2, 3, 4 and 5. Cluster 6 corresponds to Glycera

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 ASSEMBLAGES OF THE MARINE POLYCHAETE GENUS GLYCERA 

DPHLRW\SH EPLFURW\SH FPDFURW\SH

Fig. 2: Dendrogram of cluster analysis for benthic polychaetes, sediment texture and organic carbon using
Euclidean distance Ward minimum variance method.

Gray & Pearson (1982) noticed significant abundance of
polychaetes in marine sediments due to increased organic
matter. In the present case a more or less similar association
between Glycera and organic carbon in sediments was no-
ticed in all the stations studied. Brock et al. (2002) in their
studies on benthic polychaete communities of Hawaiian
waters used these organisms as indicator of organic enrich-
ment. The reduced numbers of macro Glycera recorded in
the present study may be due to the impact of bottom trawl-
ing carried out in this region causing damage to the fauna by
means of the fishing gears or exposure to predators (Thomas
et al. 2006).
The statistical analysis substantiated the profusion of
genus Glycera in all seasons in sediments rich in silt, clay,
sand and with high organic matter, tend to suggest them as a
good indicator of organic enrichment along the south-west
coast of India in coastal pollution monitoring surveys.
Obviously more detailed studies on the habitat selection and
the role of organic carbon in the life of this important genus
of marine polychaete are required.
$&.12:/('*(0(17
The authors thank Dr. N. P. Kurian, Director, Centre for Earth
Science Studies, for providing all the facilities to carry out
the present work. The financial assistance from the Ministry
of Earth Science, New Delhi through the sponsored project,
Coastal Ocean Monitoring and Prediction System
(COMAPS), is gratefully acknowledged. The help rendered
by the crew members of the research vessel and Mr. R. Shibu
(Junior Research Fellow, COMAPS) is specially remem-
bered.
5()(5(1&(6
Arvanitidis, C., Bellan, G., Drakopoulos, P., Valavanis, V., Dounas, C.,
Koukouras, A. and Eleftheriou, A. 2002. Seascape biodiversity pat-
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biogeography of the benthic polychaetes. Marine Ecology Progress
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associated with a tropical ocean outfall: Synthesis of a biomonitoring
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Fauchald, K. 1977. The Polychaete Worms Definitions and Keys to the
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Gesteira, J.L.G. and Dauvin, J.C. 2005. Impact of Aegean sea oil spill on
the subtidal fine sand macrobenthic community of the Ares-Betanzos
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Gray, J.S. 1981. The Ecology of Marine Sediments. Cambridge University
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Joydas, T.V., Jayalakshmy, K.V. and Damodaran, R. 2009. Polychaete com-
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244. In: Holme, N.A. and Mclntyre, A.D. (eds). Methods for the Study
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Ouseph, P.P., Prasanthan, V., Abhilash, P.P. and Udayakumar, P. 2009.
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west coast of India. Indian Journal of Marine Science, 23: 137-142.
Thomas, J.V., Sreedevi, C. and Madhusoodana Kurup, B. 2006. Variations
on the infaunal polychaetes due to bottom trawling along the inshore
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Udayakumar, P., Abhilash, P.P. and Ouseph, P.P. 2009. Assessment of water
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 0CVWTG'PXKTQPOGPVCPF2QNNWVKQP6GEJPQNQI[ ,661 9RO 1R SS 
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Nirmal Kumar, J. I., Yamini Verma, Rita N. Kumar and Manishita Das
P. G. Department of Environmental Science and Technology, Institute of Science and Technology for Advanced Studies
and Research (ISTAR), Vallabh Vidya Nagar-388 120, Gujarat, India

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,1752'8&7,21 anthropogenic alterations in the aquatic ecosystems (Prescot

Phytoplankton are the main primary producers and they con-
dition the structure and density of consumers as well as
hydrochemical properties of water (Harold et al. 2007). The
community varies widely in respect to taxonomic composi-
tion and cell size and their density increase with increasing
nutrient status or trophic state (Senerpont 2007).
Phytoplankton are very suitable organisms for the deter-
mination of the impact of toxic substances on the aquatic
environment because any effect on the lower level of the
food chain will also have consequence on the higher level
(Akbulut et al. 2001). Phytoplankton encountered in the
water body reflect the average ecological condition and there-
fore, they may be used as indicator of water quality (Hakason
et al. 2003). Several studies in India, have correlated
phytoplankton composition in response to water parameters
in wetlands and reservoirs under distinct trophic states.
Chattopadhyay & Benerjee (2007) observed phytoplankton
response immediately to the surrounding changes and hence,
their standing crop alters due to indicated water quality.
Knowledge of phytoplankton population dynamics is rel-
evant because temporal and spatial fluctuations in composi-
tion and biomass may be efficient indicators of natural or
2004, Olele 2008).
Studies on phytoplankton diversity and abundance in
Gujarat are sparse and restricted to mere short term taxo-
nomic observation reports. However, certain limnological
studies were done by Nirmal Kumar et al. (2005) on assess-
ment of eutrophication and weed growth of certain wetlands.
Nirmal Kumar et al. (2006, 2007, 2008) examined the pat-
terns of site-specific variation of waterfowl community,
abundance and diversity in relation to seasons in Nal Wetland
Bird Sanctuary. Nirmal Kumar (2008, 2009) assessed the
variations in hydrochemical characters of wetlands of
Gujarat. The present work assessed the composition and sea-
sonal variation of phytoplankton in relation to hydrochemistry
to understand the status of the wetland Thol Bird Sanctuary.
0$7(5,$/6$1'0(7+2'6
Site description: Thol is a shallow wetland with maximum
depth of 3 m, having an area of 6.99 sq. km and located 24
km from Ahmedabad city, district Mehsana (23°15’ to 23°30’
N and 72°30’ to 72°45’ E) of Gujarat. The wetland was de-
clared as a bird sanctuary in November 1988, which
inhabitate hundreds of bird species of which 30 bird species
are migratory. Wetland Thol is characterised by alkaline
 Nirmal Kumar J.I. et al.
nature of water and good oxygenation of surface water. For
decades the wetland has been used for irrigation purposes
and fishing (Fig. 1). Rural settlement is found towards the
north east and north west direction of the wetland. As the
wetland comes under the natural conservation area for birds
of Gujarat, the anthropogenic activities such as washing,
poaching, bathing, cattle wading and illegal entry are strictly
restricted. Birds like Common coots, Shelduck, Common
pochards, Flamingoes, Painted stork, Spoonbills, Ibis are
dominant species throughout the investigated period and
Spot Billed Duck, Eurasian Wigeon, Asian openbill are found
mostly seasonal and appeared only during in winter season.
Ipomea aquatica, Polygonum glabrum, Typha angustata, T.
domengensis, Eichhornia crassipes and Salvinia natans are
some marshy and floating aquatic macrophytic species pro-
viding food and shelter to these aquatic fowls.
Water sampling and analysis: Monthly surface water sam-
ples were collected for two years (September, 2007 to Au-
gust, 2009) for physical and chemical analysis at two sta-
tions (Stations 1 and 2). Collection was made between 7:00
and 9:00 hours IST (Indian Standard Time). Samples for dis-
solved oxygen were collected just a few centimetres below
the surface and fixed with Winkler’s reagents. The collected
samples were brought to the laboratory within 1h and stored
in a refrigerator for further analysis. The physico-chemical
analysis was made following the methods given by APHA
(2000) and Trivedy & Goel (1987). The average of two sam-
ples was considered as one reading.
Phytoplankton sampling and identification: The
phytoplankton samples were simultaneously collected by
using 20 P mesh size planktonic net along with the water
samples, and fixed by addition of 1 mL of 4 % formalin
solution. The camera lucida diagrams were drawn under light
Fig. 1: Localisation of the study sites, Thol Bird Sanctuary.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
microscope and identification of phytoplankton was made
by using various monographs, books and published litera-
ture (Desikachary 1959, Prescott 2004, Hadi et al. 1984).
5(68/76
The average hydrochemical properties of water are given in
Table 1 for the two years, and Fig. 2 shows their annual vari-
ation. Water temperature varied from 14.8 to 29.6°C and the
maximum water temperature was achieved in the summer
season of second year, and the minimum in the winter sea-
son of the same year. Values of pH remained alkaline, and
lowest value of 8.2 was observed in the monsoon season of
the first year. The dissolved oxygen fluctuated between 4.5
and 7.9 mg/L, with minimum values in May, and maximum
values (7.9 mg/L) in winters of the second year. Decomposi-
tion processes and sediment oxygen demand were sufficient
to cause lower dissolved oxygen values. In the dry period
Thol wetland had low dissolved oxygen, and high pH and
temperature, when compared to rainy and winter seasons.
Total alkalinity values varied in a narrow range from
173 to 244 mg/L. In first year alkalinity was characterized
by higher values than the second year. Chlorides attained
their maximum in post monsoon season (154 mg/L) and
found minimum in monsoon season (71 mg/L) of first year,
whereas in the second year the values of chlorides ranged
from 69 to 146 mg/L.
Nutrient analysis revealed the remarkable differences,
observed in the concentration of nitrate and phosphate after
monsoon season. Sulphate values ranged between 16 and 80
mg/L, and phosphate between 0.5 and 3.3 mg/L. The recorded
high phosphate values are probably due to release of great
amounts of runoff from the agriculture fields, and the lower
values of phosphate could be attributed to vigorous uptake
by plankton. The maximum value of nitrate was found in
summer (1.2 mg/L), and minimum in winter (0.2 mg/L ).
The highest values of nitrate reflect the direct effect of the
agriculture run off, while the lowest values are the indicator
of phytoplankton uptake. On the other hand, phosphate con-
tent was recorded maximum in summer season (3.3 mg/L ),
and minimum in monsoon season (0.4 mg/L ). Mean con-
centration of sodium (247 mg/L) and potassium (212 mg/L)
were recorded maximum in second year.
The correlation analysis of the important parameters with
the phytoplankton density showed a positive correlation of
temperature with Cyanophyta (0.17), Bacillariophyta (0.05)
and Euglenophyta (0.02), and DO with Cyanophyta (0.65).
Further, sulphate and nitrate also showed a positive correla-
tion with Cyanophyta (0.47) and Euglenophyta (0.49). Fol-
lowing a similar trend at site 2 also, DO correlated posi-
tively with Cyanophyta (0.39), Chlorophyta (0.61) and
 HYDROCHEMISTRY AND PHYTOPLANKTON COMMUNITY IN THOL WETLAND 

Table 1: Hydrochemical characteristics of water in Thol Bird Sanctuary. of the two years was compared, the highest similarity was
seen between the first and second year. According to the dia-
First Year Second Year
2007-2008 2008-2009 gram obtained from cluster analysis, similar clusters were
Mean SD SV Mean SD SV seen between different months of both the years (Fig. 4).
Temp. 21.9 4.63 21.41 22.3 4.73 22.4 ',6&866,21
pH 9.3 0.52 0.27 8.9 0.40 0.16
DO 5.6 1.12 1.25 6.4 1.09 1.20 Wetlands exhibit different water quality status depending
Total alkalinity 213 15.70 246 211 19.65 386 on the geological formation in the catchment and inflow in-
Hardness 131 17.42 303 149 16.34 267
cludes wastewater (Bendell-Young et al. 2000). It is evident
Chloride 122 28.02 785 110 25.5 651
SO4 21.0 5.12 26.18 43 19.9 398 that the maintenance of healthy conditions in aquatic sys-
PO4 0.99 0.57 0.32 1.8 0.89 0.8 tems is dependent on the hydrochemical properties of water
NO3 0.66 0.24 0.06 0.5 0.23 0.1 and biological diversity. The temperature of the water body
Na 167 49.30 2431 247 62 3817
is an important parameter influencing the water quality. In
K 133 26.70 713 212 30.04 902
both the years temperature varied according to the seasonal
All values are in mg/L except temperature (°C) and pH; Standard Devia- fluctuations of atmospheric temperature with maximum dur-
tion (SD); Standard Variance (SV) ing summers and minimum during winters (Nirmal Kumar
et al. 2005). Dissolved oxygen is an important parameter of
Bacillariophyta (0.58). Nitrate also showed a positive corre-
aquatic system, which is essential to the aerobic metabolism
lation with Cyanophyta (0.76), Chlorophyta (0.58) and
of all aquatic organisms (Wetzel 1975). In summer with the
Bacillariophyta (0.58).
increase in water temperature, there was reduction in DO,
Phytoplankton: The phytoplankton consisted of total 102 whereas in winter months due to decrease in temperature,
taxa belonging to Cyanophyta (44), Bacillariophyta (25), the level of DO increased. These results were in conformity
Chlorophyta (23) and Euglenophyta (10) (Table 3). Accord- with Ahmed Masood & Krishna Murthy (1990) and
ing to the percentage distribution of species diversity, the Srivastava et al. (2003). Comparatively low values of DO in
highest rich algal group was Cyanophyta (42.8%). In terms the first year indicate an oxygen deficient condition, which
of the counting results, the total density of Bacillariophyta, could be due to the high respiratory activity of the biota
Chlorophyta and Euglenophyta was 31.0%, 19.7% and 5.6% present there (Alom & Zaman 2006). Low quantity of water
respectively (Table 3). level during spring, summer and pre monsoon may be the
The diagram obtained from cluster analysis showed that reason for the increase of the chloride concentration which
the five different groups comprised in the water samples at corroborated with the study of Sukhija (2007). Higher val-
14% hierarchical level (Fig 4). The first important group ues of hardness were observed during summer, which may
within these was characterized by the dominance of be due to low water level and high rate of decomposition
Coelastrum cambricum and other group species Cosmarium and evaporation thus concentrating the salts (Chatterjee &
monomatum and Aphanocapsa elachista. The second im- Raziuddin 2007).
portant group was characterised by the increase of the green High concentration of nutrients like phosphate, sulphate,
algae. In this month Scenedesmus and Pediastrum from chloride, nitrate and others was recorded in second year. High
Chlorophyta were dominant species. The second group was phosphate concentration indicates fertilizer runoff, domes-
characterized by the increase of the green algae together with tic waste discharge and detergents. Similar observations were
beginning of spring and end of winter. The third group com- also made by Khare et al. (2007). The monthly variation of
prised with the decline of blue green algae, green algae and hydrochemical properties also indicated that concentration
the increase of diatoms. Cyclotella and Nitzschia from the of nutrients was greater during warmer months in the
diatoms and also Trachelomonas from Euglenoids were wetland, which could be attributed to high atmospheric tem-
prevalent species. The significant fourth and fifth group con- perature, evaporation and high amount of entry of waste-
tained only spring samples. This group was characterized discharge from surrounding villages corroborating with the
by the dominant green algae. Merismopedia and findings of Ranjan et al. (2007).
Scenedesmus species were water blooms. Cocconeis, Na- In tropical regions, different algal groups were seen as if
vicula and Nitzschia from diatoms and Anabaena, they followed typical succession model (Reynold 2004). It
Aphanocapsa, Gloeocapsa, Merismopedia and Microsystis is thought that these successions cause the variations related
from Cyanophyta were prevalent species in the same group. to the use of the light and temperatures. According to the
When the relative abundance of species in the samples number of species, Chlorophyta and Bacillariophyta type of

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Nirmal Kumar J.I. et al.
Fig. 2: Monthly variation in temperature, pH (a, b), DO, total alkalinity (c,
d), chlorides, sulphate (e, f), phosphate, nitrate (g, h) and sodium, potas-
sium (i, j) at Thol wetland from September 2007 to August 20. Values are
in mg/L except temperature (°C) and pH.
phytoplankton existed in Wetland Thol. The wetland had a
typical phytoplankton population of eutrophic wetlands,
which represent members of Chlorophyta, Bacillariophyta
and Cyanophyta along with harmful water blooms (Paerl et
al. 2001). The seasonal succession of algae in Wetland Thol
was Chlorophyta and diatoms in the spring, Chlorophyta in
early summer, Cyanophyta in late summer and Diatoms in
autumn and winter. Generally, it was a complicated succes-
sion, as it happens in many shallow tropical wetlands
(Hutcinson 2007).
In the dendrograms obtained from cluster analysis, spe-
cies making blooms encountered as densely composed
groups. Monoraphidium sp., Oocystis borgei, Pediastrum
boryanum and Secenedesmus sp. showed blooming some-
times. Round (2002) reported that some Chlorococcales
members are quite abundant in tropical wetlands showing
transition from oligotrophic condition to eutrophic one. In
terms of Hutchinson (2007), these species are dominant
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
Fig. 3:Density in percentage of phytoplankton groups.
Fig. 4: Dendrograms for hierarchical clustering of the samples, months
and phytoplankton groups in the Wetland Thol.
organisms in eutrophic waters. Wetland Thol is located in
sensum environment; green algae are dominant and are re-
sponsible for making blooms in some months. Also,
Chroococcus sp. from blue green algae and Euglena graci-
lis from euglenoids were prevalent species in the wetland
having eutrophic properties in particular. Prescott (2004)
reported that Cyanophyta members make blooms in the stag-
nant water of certain tropical wetlands. In the eutrophication
of wetland ecosystems, the blooming of blue greens is a fre-
quent event (Moss et al. 2006). Blue green algae are the most
prevalent and harmful for people and limit the convenient
use of water (Pitois et al. 2001).
When the phytoplankton dynamics expressed to seasons,
blue green algae were found to make blooms in summer and
early autumn. Cryptomonas genus is an indicator of eutrophic
wetlands (Akbulut & Yildiz 2001). In these months, blue
greens and green algae made blooming in phytoplankton.
Anabaena catenula, Microcystis aeruginosa, Nodularia
spumigena and Pseudoanabaena limnetica species were
noticed important increases in summer and late autumn.
Because of organic pollution, Euglenoid members were
often found in Wetland Thol. Phacus and Trachelomonas
from Euglenophyta were dominant organisms in spring
months. It is reported that Euglenophyta members generally
develop very well in waters which is rich in organic sub-
stances (Round 2003).
 HYDROCHEMISTRY AND PHYTOPLANKTON COMMUNITY IN THOL WETLAND 

Table 2: Hydrochemical correlation coefficient analysis of water, Thol Bird Sanctuary, year 2007-2009.

Temp pH DO Tl.alk Cl SO4 PO4 NO3 Na K Chloro- Cyano Bacillario Eugleno

phyta phyta phyta phyta

Temp 1
pH 0.16 1
DO -0.71 -0.32 1
Tl.Alk 0.57NS 0.42 -0.56 1
Cl 0.08** 0.79 -0.16 0.07NS 1
SO4 0.31 0.19 0.05* 0.26 0.29 1
PO4 0.64 0.37 -0.31 0.60 0.30 0.75 1
NO3 -0.67NS 0.29 0.55NS -0.46 0.43 -0.20 -0.34 1
Na 0.37 0.31 -0.06 0.57 0.22 0.82 0.86 -0.23 1
K 0.29 -0.13 0.08* 0.41 -0.19 0.74 0.71 -0.44 0.83 1
Chlorophyta -0.87 0.06 0.65 -0.43 0.19 -0.04 -0.36 0.72 -0.12 -0.08 1
Cyanophyta 0.17 0.17 -0.23 0.09 0.29 0.47 0.37 -0.18 0.24 0.20 -0.07 1
Bacillariophyta 0.05* -0.54 -0.10 0.16 -0.76 -0.22 -0.06 -0.41 -0.03 0.24 -0.30 -0.05 1
Euglenophyta 0.02* 0.16 -0.05 0.03* 0.29 0.49 0.35 -0.04 0.30 0.25 0.10 0.93 -0.10 1

Level of significance: ** = (P d 0.01), * = (P d 0.05), NS = Non-significant (P > 0.05).

Table 3: Number of phytoplankton and mean density (units × 105 per litre) different algal groups as evident from the positive correla-
of the major taxonomic groups to total phytoplankton in Wetland Thol
tion coefficient values. Phosphate seems to limit the growth
Bird Sanctuary in both the investigated years.
as depicted from the negative correlation coefficient values
Group Number Mean density Total in for both the years with Chlorophyta and Bacillariophyta den-
of Species Year Percent sity. Ersanli et al. (2003) stated that temperature, pH, alka-
2008 2009
linity and phosphate have been emphasized to be limiting
Cyanophyta 44 4.1976 5.6848 42.8 factors for controlling distribution of Cyanophyceae which
Chlorophyta 25 1.826 2.7192 19.7 also corroborated with the present study. Tripathy & Panday
Bacillariophyta 23 2.86 4.5276 32.0 (1990) and Rana & Nirmal Kumar (1993, 2005) reported
Euglenophyta 10 0.4796 0.8184 5.6 that high water temperature, phosphate, nitrate and low DO
Total 102 9.3632 13.75 100
support the growth of Chlorophyceae. pH was found to be
in the alkaline range (8.9 to 9.3) supporting a good popula-
The results obtained from cluster analysis and the count- tion of the diatoms. Moreover, DO content was found to be
ing methods supported to each other regarding phytoplankton considerably high in colder months. The plankton commu-
abundance. There was no difference between the nity, on which the whole aquatic population depends, is
phytoplankton composition and the seasonal dynamics ex- largely influenced by interaction of a number of limiting
cept for some months. When the diagram was examined, the factors and number of hydrochemical and biological factors
dominant species comprised groups in cluster analysis of acting simultaneously must be taken into consideration in
both the years. It was reported that these taxa composed of understanding the diversity of plankton population. The data
groups in wetland with eutrophic characteristic (Hutchinson obtained from this study indicated that the wetland is pro-
2007). gressing to eutrophication stage.
Diversity of plankton population is fairly dependent on
5()(5(1&(6
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M. K. Jain, L. K. Dadhich and S. Kalpana
P.G. Department of Chemistry, Govt. College, Kota, Rajasthan, India

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India is rich in water resources being endowed with a criss-
cross network of rivers that can meet a variety of water re-
quirements of the country. Due to need for meeting the in-
creasing demands by urbanization, industrialization and
modern agricultural activities, the available water resources
are getting depleted and the water quality has deteriorated.
The Indian rivers are polluted due to discharge of untreated
sewage, industrial effluents and agricultural runoff. In the
country maximum precipitation occurs in four months of
monsoon period and rivers also have maximum flow in this
period. In lean seasons rivers have very little or no flow. To
meet the water demand for drinking, domestic, agriculture
and industry in lean periods, several types of water storage
devices like anicuts, check dams, small and large dams, etc.
have been constructed across the country on main course of
river streams. The water quality of these storage structures
depends upon physiography, geology, geomorphology,
hydrogeology, demography, land use pattern and human set-
tlement of the area concern.
Several workers have studied water quality of storage
structures all over the world. Water quality studies of dams
worldwide in last fifty years have been reviewed by Biswas
& Tortajada (2001). Ganesan et al. (2004) studied seasonal
variation in the water quality of Pilavakkal reservoir in West-
ern Ghats in India with respect to biological and chemical
properties. Water quality of inflow and outflow of Mettur
dam, constructed over River Cauvery, has been studied by
Mathivanan et al. (2005) who reported high pollution load
attributed to dense urbanization of the area. Karibasappa et
al. (2009) have studied eutrophication in Hosur town lakes
and reported its high level due to contamination with sew-
age and agriculture runoff. Singh et al. (2005) studied the
water quality of six reservoirs of Damodar river basin. Wa-
ter quality of Pench reservoir was studied by Pokale et al.
(2010) for drinking and agricultural activities. Considering
the impacts of agricultural runoff, and domestic and indus-
trial effluents on water quality of rivers and dams, the present
study was undertaken to investigate water quality of
Kishanpura dam for agricultural activities.
7+(678'<$5($
Fern leaf shape River Parbati, a principal tributary of River
Chambal, is situated at east longitudes of 75°22’ to 77°12’
and north latitudes of 24°19’ to 25°51’. It occupies a total
catchment area of 15,861 km 2 and originates in the
Vindhyachal ranges at an elevation of 609 m near Astha town
in Sehore district of Madhya Pradesh. The river enters
Rajasthan near Khurai village in Baran district and joins
River Chambal near Pali village of Kota district, after trav-
elling 159 km. The river is joined by a number of tributaries
in Rajasthan, the more important of which are Uperni, Dubraj,
Andheri, Beram, Kosam, Ahelil and Sukni.
The Kishanpura dam is constructed across River Parbati
just downstream of the joining point of River Andheri near
Atru town of Baran district. The dam was constructed in
1882. Catchment area of the dam is 10443 km2 with an aver-
age annual rainfall of 848.50 mm. The technical data are not
available but the data, engraved on a stone fixed on dam,
read as: constructed in S.B.T. 1941, length 1310 ft, width 13
ft, canal length 36 miles, cill 6 ft below anicut, area com-
manded 30,000 bighas average. The dam is also called pick-
up weir. The dam height was raised by 0.73 m in 1960 and
again 0.30 m in 2004. Channels of 0.45 m were also fitted in
2004 by which temporarily fencing is done in monsoon pe-
riod with the help of iron shutters. The dam is broad crested

vertical drop weir constructed in stone masonry with lime
mortar with wing walls and earthen flanks on both sides,
without any protection works in upstream and down stream.
By construction of the dam, the river water is stored in
river channel which is diverted to canals by sluice gates.
Canal system presently nourishes 12500 acres land of 58
villages by 58 km long main canal, 17 km Rajwah branch
and 88 km minor and distributaries in Atru, Baran and
Mangrol tehsils. No Kharif irrigation was done earlier than
1960 and canal used to be opened for Rabi crops in October.
After 1960 the water was also provided for Kharif which is
possible by increased height of dam.
Irrigated agriculture is dependent on an adequate water
supply of usable quality. In irrigation water evaluation, em-
phasis is placed mainly on the chemical and physical char-
acteristics of the water. Here attempt has been made to as-
sess the irrigation water quality of Kishanpura dam in Baran
district of Rajasthan. Since, for more than a century, water
of the dam is being used for irrigation, it is worthwhile to
assess suitability for its intended use. The quality character-
istics studied in the present investigations were pH, electri-
cal conductivity (EC), total dissolved solids (TDS), total
M. K. Jain et al.
hardness (TH), calcium, magnesium, potassium, sodium, car-
bonate and bicarbonate. The calculated indices were sodium
percentage (SP), sodium adsorption ratio (SAR), residual
sodium carbonate (RSC), exchangeable sodium percentage
(ESP) and permeability index (PI).
0$7(5,$/6$1'0(7+2'6
Water samples were collected for three years (January 2008
to September 2010) in the three seasons i.e., winter,
premonsoon and postmonsoon. In order to get truly repre-
sentative sample integrated sampling was done. Samples
were drawn from six different identified locations and then
mixed together. Samples were collected in precleaned
polypropylene screw capped bottles of one litre capacity.
The standard methods and procedures were used for quanti-
tative estimation of water quality parameters. Determination
of pH was carried out at site. Samples were brought to the
laboratory to analyse other physico-chemical parameters
using standard methods (APHA 1995).
From the water quality data, the irrigational indices i.e.,
sodium percentage (%Na), sodium adsorption ratio (SAR),
residual sodium carbonate (RSC), exchangeable sodium per-
centage (ESP) and permeability index (PI) were calculated
from the following equations. For calculation of these indi-
ces, concentrations are taken in meq/L.
Magnesium content: Magnesium content of water is con-
sidered as one of the most important qualitative criteria in
determining quality of water for irrigation. Magnesium haz-
ard ratio is calculated by the following formula (Pitchaiah
1995).
Mg content = [Mg2+/ (Mg2 ++ Ca2+)] 100
Sodium percentage (%Na): Suitability of water for irriga-
tion is evaluated by sodium percentage. % sodium is calcu-
lated by following equation (Chopra & Kanwar 1999).
Na% = [ Na+/ (Ca2++ Mg2++ Na+ + K+)] 100
Sodium adsorption ratio (SAR): Sodium adsorption ratio
is an important parameter to determine the suitability of ir-
rigation water and is calculated by the following formula
(Wilcox 1955).
SAR = Na+ /[(Ca2+ + Mg 2+)/2]1/2
Exchangeable sodium percentage (ESP): It is a degree of
saturation of the soil exchange complex with sodium. It is
calculated by the following relationship (Dwivedi & Pathak
2007).
100 (-0.0126 + 0.01475 SAR)
ESP = ––––––––––––––––––––––––––
1 + (-0.0126 + 0.01475 SAR)
Residual sodium carbonate (RSC): The residual sodium
)LJ.LVKDQSXUDGDPSLFNXSZHLURQ3DUEDWLULYHU carbonate (RSC) is used for evaluating high carbonate wa-

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 WATER QUALITY ASSESSMENT OF KISHANPURA DAM IN BARAN, RAJASTHAN 

Table 1: Calculated indices and water quality data of Kishanpura dam.

Parameters 2008 2009 2010

Winter Pre- Post- Winter Pre- Post- Winter Pre- Post-
monsoon monsoon monsoon monsoon monsoon monsoon

Physico-chemical Parameters
pH 7.70 7.95 7.80 7.95 8.25 8.05 8.10 8.20 8.05
EC, µS/cm 360 382 282 348 378 260 334 370 284
TDS, mg/L 227 241 178 219 238 164 210 233 179
CO32-, mg/L as CaCO3 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
HCO3-, mg/L as CaCO3 122 139 106 128 148 114 118 126 112
TH, mg/L as CaCO3 122 135 92 118 132 96 114 128 104
Ca2+, mg/L 26.8 30.0 18.8 23.2 28.8 20.4 25.6 27.2 19.6
Mg2+, mg/L 13.2 14.4 10.8 14.4 14.4 10.8 12.0 14.4 13.2
Na, mg/L 14.2 17.6 12.8 13.9 18.7 11.9 15.2 19.4 10.7
K, mg/L 2.1 2.7 1.8 2.3 3.2 1.9 2.1 2.7 1.6
Calculated Indices
%Na 19.84 21.65 22.78 19.99 23.00 20.81 22.07 24.29 17.99
SAR 0.56 0.66 0.58 0.56 0.71 0.53 0.62 0.75 0.46
ESP -0.44 -0.29 -0.41 -0.44 -0.22 -0.48 -0.35 -0.16 -0.59
RSC 0.00 0.08 0.28 0.20 0.32 0.36 0.08 -0.04 0.16
PI 381.46 362.32 452.83 402.05 375.86 459.28 391.84 354.59 434.08

ters. RSC is calculated by the formula given below (Richards
1954).
RSC = (CO32- + HCO3 -) – (Ca2+ + Mg 2+)
Permeability index (PI): Permeability index is calculated
by the method given by Domenico & Schwartz (1990). PI is
used to evaluate the sodium hazards of irrigation water.
PI = [( Na+ + HCO3 -) /( Ca2+ + Mg 2+ + Na+)] 100
5(68/76$1'',6&866,21
The results obtained from analysis of water samples of
Kishanpura dam and calculated indices are given in Table 1.
Total dissolved salts (TDS): Water used for irrigation can
vary greatly in quality depending upon type and quantity of
dissolved salts. In dam water salts are present in relatively
small but in significant amounts. The salts originate from
dissolution or weathering of the rocks and soil, including
dissolution of lime, gypsum and other slowly dissolved soil
minerals. The dissolved salts remain behind in soil as water
is used up by crops or evaporates. Salinity problems exist if
salt accumulates in root zone to a concentration that causes a
significant loss in yield. This occurs when salts accumulate
in root zone to such an extent that the crop is no longer able
to extract sufficient water from the salty soil solution, re-
sulting in a water stress for a significant period of time. If
water uptake is appreciably reduced, the plant growth rate is
also reduced. Water with TDS less than 450 mg/L is consid-
ered good, and that with greater than 2000 mg/L is unsuit-
able for irrigation (Westcott & Ayers 1984). In the present
study values of TDS ranged from 164 mg/L to 241 mg/L.
The minimum value was observed in postmonsoon 2009,
and maximum in premonsoon 2008.
Electrical conductivity (EC): The most significant water
quality guideline on crop productivity is the water salinity
hazard as measured by electrical conductivity. The primary
effect of high EC water on crop productivity is the inability
of the plant to compete with ions in soil solution for water.
Plants can only transpire water and hence usable plant water
in soil solution decreases dramatically as EC increases, even
though the soil may appear wet. Water with EC less than
250 µS/cm is considered good and greater than 3000 µS/cm
is unsuitable for irrigation (Westcott & Ayers 1984). In
present investigation EC ranged from 260 µS/cm to 382
µS/cm. The minimum value was observed in postmonsoon
2009, and maximum in premonsoon 2008.
Magnesium content (MC): Magnesium content of water is
considered as one of the important qualitative criteria in de-
termining the quality of water for irrigation. Generally, cal-
cium and magnesium maintain a state of equilibrium in most
waters. More magnesium in water will adversely affect crop
yields as the soils become more alkaline. In the present study,
the magnesium content of the water of Kishanpura dam var-
ied from 10.8 mg/L to 14.4 mg/L. The dam water is suitable
for irrigation purpose in terms of magnesium content.
Sodium percent (% Na): Sodium percent is another impor-
tant factor to study sodium hazard. It is calculated as the
percentage of sodium and potassium against all cationic con-
centration. Sodium reacts with soil to reduce its permeabil-
ity. The use of high percent sodium water for irrigation stunts

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 M. K. Jain et al.
the plant growth. The most suitable for irrigation is that water
having % Na values less than 20%. When % Na is greater
than 20%, permeability will be reduced (Wilcox 1955). The
finer the soil texture with high organic matter content, the
greater the impact of sodium on water infiltration and aera-
tion. The calculated sodium percent values of Kishanpura
dam ranged from 17.99 to 24.29. Normally, gypsum is added
to soil to reduce the effect of high percentage of sodium in
irrigation water.
Sodium adsorption ratio (SAR): Excess sodium in water
used for irrigation change soil properties and reduces soil
permeability. Hence, the assessment of sodium concentra-
tion is necessary while considering the suitability for irriga-
tion. The degree to which irrigation water tends to enter into
cation-exchange reactions in soil can be indicated by the
sodium adsorption ratio. Sodium replacing adsorbed calcium
and magnesium is a hazard as it causes damage to the soil
structure, which becomes compact and impervious. Water
with SAR from 0 to 10 is considered good and with greater
than 26 is considered unsuitable for irrigation. In the present
study SAR values ranged from 0.46 to 0.75 showing the
water of excellent category. It can be used for irrigation on
almost all types of soil.
Exchangeable sodium percentage (ESP): It is a degree of
saturation of the soil exchange complex with sodium. In study
of successive three years, all the calculated values of ESP
were in negative showing that the water quality is suitable
for irrigation.
Residual sodium carbonate (RSC): Bicarbonate and car-
bonate concentration of water affects its suitability for irri-
gation. Water with high concentration of bicarbonate and
carbonate has a tendency of calcium and magnesium to pre-
cipitate, as the water in soil becomes more concentrated as a
result of evaporation and plant transpiration. They get fixed
in the soil by the process of base exchange, thereby decreas-
ing soil permeability. The calcium and magnesium precipi-
tate as carbonates and residual carbonate and bicarbonates
are left in solution as sodium carbonate. The water with high
RSC has high pH and land irrigated with such water becomes
infertile owing to deposition of sodium carbonate; as known
from black colour of the soil. According to U.S. Salinity
Laboratory, an RSC value less than 1.25 meq/L is safe for
irrigation. A value between 1.25 and 2.5 meq/L is of mar-
ginal quality, and value of more than 2.5 meq/L is unsuit-
able for irrigation. In the present study RSC values are be-
low 1.25 meq/L in all the three years. So water of Kishanpura
dam can be considered safe for irrigation.
Permeability index (PI): The soil permeability is affected
by long term use of irrigation water. Sodium, calcium, mag-
nesium and bicarbonate content of the soil influence it.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
Domenico & Schwartz (1990) evolved a criterion for assess-
ing the suitability of water for irrigation based on the perme-
ability index. Accordingly, waters can be classified as Class I,
Class II and Class III orders. Class I and Class II waters are
categorized as good for irrigation with 75% or more maxi-
mum permeability. Class III waters are unsuitable with 25%
of maximum permeability (Joshi et al. 2009). In the present
study, the minimum value of PI is 354.59, hence, the water
quality of Kishanpura dam is good for irrigation.
&21&/86,21
In the present study, EC, TDS, MC, % Na, ESP, RSC, SAR
and PI values were found well within the permissible range,
hence, the water quality of Kishanpura dam is good with
respect to irrigational use.
5()(5(1&(6
APHA, AWWA, WPCF 1995. Standard Methods for Examination of Wa-
ter and Wastewater. , 19th Edn., American Public Health Association,
Washington, DC., U.S.A.
Biswas, A.K. and Tortajada, C. 2001. Development of large dams: A glo-
bal perspective. Int. J. Water Resour. Devel., 17: 9-21.
Chopra, S.L. and Kanwar, J. S. 1999. Analytical Agricultural Chemistry.
Kalyani Publishers, New Delhi.
Domenico, P.A and Schwartz, F.W. 1990. Physical and Chemical Hydrol-
ogy. John Wiley and Sons, New York.
Dwivedi, S.L. and Pathak, V. 2007. Studies of water quality of Mandakini
river in Chittrakoot for irrigation purposes. Ind. J. Env. Prot., 8:
751-754.
Ganesan, M., Jayabalan, N. and Jegatheesan, K. 2004. Environmental sta-
tus and seasonal variation in the water quality studies of Pilavakka
reservoir at Western Ghats in India. J. Plant Archives, 4: 195-201.
Goel, P.K. 2006. Water Pollution: Cause, Effects and Control. Second Re-
vised Edition, New Age International Publishers, New Delhi.
Joshi, Dhirendra Mohan, Kumar, Alok and Agrawal, Namita 2009. As-
sessment of the irrigation water quality of River Ganga in Haridwar
District. Rasayan J. Chem., 2(2): 285-292.
Karibasappa, H., Arvinda, H.B. and Manjappa, S. 2009. A study on
eutrophication level in Hosur town lakes. Nat. Env. Poll. Tech., 8(2):
297-300.
Mathivanan, V., Vijayan, P. and Sabhanayakam, S. 2005. Pollution stud-
ies on River Cauvery in Mettur, Tamil Nadu, J. Expt. Zoology, 8:
321-328.
Pitchaiah, P.S. 1995. Ground Water. Scientific Publishers, Jodhpur,
Rajasthan.
Pokale, W.K., Thakur, J.N. and Warhate, S.R. 2010. Water quality status of
Pench Reservoir (India). J. Environ. Science & Engg., 52(3): 255-258.
Richards, L.A. 1954. Diagnosis and improvement of saline and alkali soils.
In: Agriculture Handbook, U.S. Department of Agriculture, pp. 60.
Singh, A.K., Mondal, G.C., Singh, P.K., Singh, S., Singh, T.B. and Tewary,
B.K. 2005. Hydrochemistry of reservoirs of Damodar river basin, In-
dia: Weathering process and water quality assessment. J. Environ. Ge-
ology, 48: 1014-1028.
Westcott, D.W. and Ayers, R.C. 1984. Water Quality Criteria in Irrigation
with Reclaim Municipal Wastewater: A Guidance Manual. Edited by
G.S. Pettygrobe and T. Asano, State Water Resources Control Board
Sacramento, California.
Wilcox, L.V. 1955. Classification and Use of Irrigation Water. U.S. Deptt.
of Agric., Circular 969.
 0CVWTG'PXKTQPOGPVCPF2QNNWVKQP6GEJPQNQI[ ,661 9RO 1R SS 
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M. P. Subin and V.A. Aneesha
Department of Botany, Sree Narayana College, Nattika-680 566, Distt. Thrissur, Kerala, India

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,1752'8&7,21
The poorer segments of the population, both in urban and
rural areas, generally use well water as a main source of drink-
ing water. It is also noticed that when the public water sup-
ply fails, the well water further serves as a source of direct
drinking water for even the rich section of population. Stud-
ies on water quality properties are therefore necessary to de-
termine the extent of pollution so as to monitor likely dan-
ger to the human population. The tremendous organic loads
imposed by urban sewage and other wastes constitute a ma-
jor cause of pollution of natural water bodies (Hynes 1960).
In view of the public apprehension of the hazards of water
pollution, regular water quality monitoring of water bodies
is highly necessary (Renn 1970). The test result allows us to
find out whether the water we drink and use for household
activities is reliable and safe. In light of this information, the
authors have tried to study the quality of well waters col-
lected from various areas of Ernakulam district to see whether
these waters are fit for human consumption or not.



during the month of March 2010. The location and source
of water samples are given in Table 1.
Analysis of water samples: The samples collected were
brought to the laboratory and the parameters like pH, elec-
trical conductivity (EC) and total dissolved solids (TDS)
were measured immediately after collection. Other physico-
chemical parameters like colour, odour, taste, turbidity, to-
tal hardness (TH), total alkalinity, biochemical oxygen de-
mand (BOD), dissolved oxygen (DO), calcium, chlorides,
magnesium, sulphates, iron and nitrates were analysed within
36 hrs of collection. The standard methods were adopted for
the analysis of water samples (APHA 1995).
Comparison with BIS standards: Physico-chemical param-
eters analysed and measured on water samples were com-
pared with BIS (1998) for drinking water. Each data pro-
vided in the Table 2 is an average of three samples collected
from each area under study.








Sample collection: Samples of home well waters and the
water supplied by Kerala Water Authority (KWA) through
pipeline in Ernakulam district were collected in high grade
plastic bottles of one litre capacity. Before collection, the
bottles were rinsed with distilled water and then thrice with
the well water sample. During collection, care was taken to
avoid the trapping of air within the bottle by completely im-
mersing the bottle within the water sample until the bottle is
completely filled in with the water sample. The samples were
collected from some selected regions of Ernakulan district
The results of water analysis of the collected samples are
presented in Table 2. It is clear that with the exception of
W4 sample, all the other samples were colourless, whereas
W4 was light brown coloured. The excess iron present in
water can cause the formation of insoluble hydroxides, which
settle out as rust-coloured silt which imparts an objection-
able reddish-brown colour and taste to water (Cohen et al.
1960). Regarding the taste, the samples W1, W2, W5 and
W6 were tasteless whereas the samples W3 and W4 were
slightly salty. There is report that at concentrations above
the aesthetic objective, chloride content imparts undesirable
tastes to water (WHO 1984). All the six water samples
 M.P. Subin and V.A. Aneesha

Table 1: Location and source of water samples collected from Ernakulam of TDS in drinking water prescribed by BIS is 500 mg/L
District.
and 1000 mg/L respectively. The water samples W1, W2,
Sl.No. Sample Code Location Source W5 and W6 have an acceptable value. The water sample W3
has a TDS value greater than the desirable limit but it is within
1 W1 Keezhumadu Home Well the excessive limit whereas W4 has a TDS value higher than
2 W2 Muttom Home Well
the maximum excessive limit. The most important aspect of
3 W3 Manjummal Home Well
4 W4 Irumpanam Home Well TDS with respect to drinking water quality is its effect on
5 W5 Kakkanad Home Well taste (Bruvold & Pangborn 1966). The drinking water con-
6 W6 KWA Water Pipe Line taining more than 500 mg/L of TDS is not considered desir-
able (Sastry & Rahee 1988) and it can also cause excessive
analysed were found to be odourless. scaling in water pipes, water heaters, boilers and household
With respect to the pH of water samples, a variation from appliances (Tihansky 1974). In this respect the present data
4.96 to 7.46 was observed. According to BIS (1998), the reveal that water samples collected from Manjummal and
permissible range of pH is from 6.5 to 8.5. In the present Irumpanam are considered to be undesirable and unaccept-
study, the pH of well water samples W1, W2 and W5 was able respectively. But in areas of acute shortage of water,
below the permissible limit, whereas of others within the people are forced to use water having TDS value above the
permissible limit. The low pH values of waters may be be- permissible limit and in such cases it should be used only
cause of the leaching of organic acids from decaying veg- after removing the excess TDS using technologies such as
etation or may be because of the presence of dissolved car- reverse osmosis and electrodialysis (Clark et al. 1977).
bon dioxide generated by bacteriological oxidation (Sawyer The concentration of iron (as Fe) in different water sam-
& McCarty 1967). Water having a pH below 6.5 can cause ples varied from 0.25 mg/L to 1.58 mg/L. The highest iron
considerable damage to the water supply system, resulting concentration was identified in the sample W4 (1.58 mg/L).
from complex interactions between pH and other physico- This value lies above the maximum excessive limit for con-
chemical parameters (Tihansky 1974). As the pH is related sumable water. Other water samples like W1, W3 and W5
to a variety of different parameters, it is not possible to de- also were identified to have iron concentration above the
termine whether pH has a direct relationship with human desirable limit but within the excessive limit, whereas W2
health but it is argued that pH has an indirect effect as it can and W6 samples have safe concentration. The ingestion of
affect water treatment processes (Aramini et al. 2009). large quantities of iron can result in haemochromatosis, a
The values of TDS in different water samples varied from condition in which normal regulatory mechanisms do not
70 to 1210 mg/L. The desirable and maximum excessive level operate effectively, leading to tissue damage as a result of
Table 2: Physico-chemical characteristics of water samples collected from various locations in Ernakulam district for comparison with BIS standards(1998)
for drinking water consumption.

Sl. No. Paramet er BIS (1998) standards Water samples

DL EL W1 W2 W3 W4 W5 W6

1 Colour Colorless - Colorless Colorless Colorless Light coloured Colorless Colorless

2 Odour Agreable - Agreable Agreable Agreable Agreable Agreable Agreable
3 Turbidity 5 10 2 1 1 2.30 0.90 0.80
4 TDS 500 1000 80 80 720 1210 110 70
5 pH 6.5-8.5 NR 6.24 5.44 7.46 7.02 4.96 7.45
6 TH 300 600 11.76 14.70 118.16 335.20 14.70 11.76
7 Calcium 75 200 2.35 3.52 42.33 78.92 3.52 3.52
8 Magnesium 30 100 1.42 1.42 12.99 35.24 1.42 0.71
9 Iron 0.3 1.0 0.86 0.25 0.38 1.58 0.42 0.27
10 Chlorides 250 1000 19.2 28.80 258.80 371.10 24.9 11.5
11 Sulphates 200 400 6.29 11.72 33.36 89.31 4.91 5.43
12 Nitrates 45 NR 1.89 2.51 1.56 1.32 3.16 1.55
13 EC - 1400 210 280 1470 1895 320 130
14 Total Alkalinity 200 600 54.90 7.63 146.40 134.20 3.05 61.0
15 BOD - 5 1.84 2.76 5.16 5.76 4.60 1.84
16 DO - - 7.6 7.2 6.4 4.8 7.1 7.7
17 Taste Tasteless - Tasteless Tasteless Salty Salty Tasteless Tasteless

Desirable limit (DL); Excessive limit (EL)

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PHYSICO-CHEMICAL PROPERTIES TO ASSES WELL WATER QUALITY 

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 M.P. Subin and V.A. Aneesha
the accumulation of iron (Dillman et al. 1987). In addition
to this, when the iron concentration in the domestic water
supplies exceeds 0.3 mg/L, it becomes objectionable for a
number of reasons, which are indirectly related to health
(Cohen et al. 1960).
A considerable variation was observed in the values of
electrical conductivity. The EC value varied from 130 to 1895
µmhos/cm. With reference to the BIS, the maximum per-
missible limit is 1400 µmhos/cm. The water sample with the
least EC value was noticed in W6, whereas the maximum in
W4. With the exception of water samples W3 and W4, all
the samples have an EC value within the permissible limit.
The higher EC values of samples W3 and W4 may be due to
higher concentration of TDS and ionized substances present
(Aramini et al. 2009). It clearly indicates that sample W3
and W4 are unfit for human consumption. Arutchelvan et al.
(2004) found significant linear relationship between EC-
TDS. The present investigation is also in support of these
findings. Similarly, it is also reported that a high positive
correlation exists between EC and chloride contents of wa-
ter (Govindaradjane et al. 2007).
Chloride content of water is another important param-
eter to be considered to know the quality of water as its higher
concentrations can impart undesirable taste to water and may
cause corrosion in the distribution system (McConnell 1972).
An aesthetic objective of 250 mg/L has been established as
maximum desirable limit and 1000 mg/L as maximum ex-
cessive limit for chloride in drinking water (BIS 1998). The
study shows that water samples W3 and W4 have chloride
values above the desirable limit but within the excessive
limit. The chloride content of all the other water samples
was under desirable limit. As chloride is highly soluble in
water, it is not easily removable; a removal of 87 % has been
reported using granular activated carbon adsorption and re-
verse osmosis (Regumathan et al. 1983).
The BOD value ranged from 1.84 to 5.76 mg/L. As per
BIS, the maximum permissible limit is 5 mg/L. The study
shows that the BOD value for water samples W3 and W4
was higher than the permissible limit. Higher BOD values
of water samples W3 and W4 clearly indicate pollution and
may be attributed to the percolation of wastewaters loaded
with biodegradable compounds (Pitchammal et al. 2009).
It is important to consider the DO content of drinking
water because oxygen imparts a good taste to water and is
an absolute requirement for the metabolism of aerobic or-
ganisms in water bodies. Decrease in DO can favour anaero-
bic decomposition of organic wastes (Sallae 1974). In the
present study, dissolved oxygen value of water samples
ranged from 4.8 to 7.7 mg/L. Water sample W4 exhibited
the least DO content, whereas the maximum in W6.
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The turbidity values of water samples ranged between
0.80 and 2.30 NTU and were within the desirable limit. The
WHO (1984) establishes that the turbidity of drinking water
should not be more than 5 NTU, and should ideally be be-
low 1 NTU. In this respect the water samples W1 and W4
were slightly turbid. In case where drinking water is turbid,
it is essential to eliminate the turbidity for effectively disin-
fect it for drinking purposes.
The total hardness of water samples varied from 11.76
to 335.20 mg/L. Water sample W4 exhibited a hardness level
above the desirable limit but within excessive limit. All other
water samples exhibited a value within the desirable limit.
Hard water causes incrustation in distribution systems and
excessive soap consumption (Coleman 1976).
The values of calcium and magnesium in water samples
varied from 2.35 to 78.92 mg/L and 0.71 to 35.24 mg/L re-
spectively. The water sample W4 has the maximum value in
both the cases and it was greater than the desirable limit but
within the excessive limit. All other water samples have ac-
ceptable value. Venkata et al. (2006) reported high positive
correlation between TDS-Mg, TH-Ca and TH-Mg. The
present study also supports this observation as there is a posi-
tive correlation between the values of magnesium, calcium,
TDS and total hardness in sample W4. With respect to sul-
phate and nitrate concentration and total alkalinity, all the
water samples W1, W2, W3, W4, W5 and W6 analysed were
well within the desirable limits of BIS.

The present investigation indicates that all the home well
waters W1, W2, W3, W4 and W5 tested for the 17 param-
eters have at least one or two water quality problems with
reference to the BIS (1998). In water sample W2, the pH
value alone deviates from the BIS whereas in water samples
W1 and W5 both pH and iron concentration are not agree-
able. In the case of water samples W3 and W4 several pa-
rameters were found to be varying with respect to BIS. Only
the KWA water is satisfying all the parameters considered
here for the study. Therefore, it is recommended that waters
from these wells are to be used for drinking purpose only
after pretreatment like filtering, boiling, reverse osmosis,
electrodialysis, etc.
0
APHA 1995. Standard Methods for the Examination of Water and
Wastewater. 20th edition, American Public Health Association, Ameri-
can Water works Association, Water Environment Federation, Wash-
ington DC.
Aramini, J.M., McLean, M., Wilson, J., Holt, J., Copes,R., Allen, B. and
Sears, W. 2009. Drinking Water Quality and Healthcare Utilization
for Gastrointestinal Illness in Greater Vancouver. Environmental and
Workplace Health Reports and Publications.
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Arutchelvan, V., Kanakasabai, V., Elangovan, R. and Nagarajan, S. 2004.
Physico-chemical characteristics of wastewater from bakelite manu-
facturing industry. Indian Journal of Environ. and Ecoplan., 8(3):
757-760.
BIS 1998. Indian Standard Specification for Drinking Water, IS: 10500,
Bureau of Indian Standards, New Delhi.
Bruvold, W.H. and Pangborn, R.M. 1966. Rated acceptability of mineral
taste in water. J. Appl. Psychol., 50(1): 22.
Clark, J.W., Viessman, W. and Hammer, M.J. 1977. Water supply and pol-
lution control. 3rd edition, Harper & Row Publishers, New York.
Cohen, J.M., Lamphake, L.J., Harris, E.K. and Woodward, R.L. 1960. Taste
threshold concentrations of metals in drinking water. J. Am. Water
Works Assoc., 52: 660.
Coleman, R.L. 1976. Potential public health aspects of trace elements and
drinking water quality. Ann. Okla. Acad. Sci., 5: 57.
Dillman, E., Gale, C., Green, W.E., Johnson, D.G., Mackler, B. and Finch,
C. 1987. Hypothermia in iron deficiency due to altered trycodo-thyro-
nine metabolism. Am. J. Physiol., 2.
Govindaradjane, S., Sundararajan, D., Sivamoorthy Reddy, S. and
Sivasankaran, M.A. 2007. Physico-chemical characteristic of waste
water from a pharmaceutical industry. Poll.Res., 26(1): 131-133.
Hynes, H.B.N. 1960. The Biology of Polluted Waters. Liverpool Univer-
sity Press, Liverpool, 202 pp.
McConnell, H.H. and Lewis, J. 1972. Add salt to taste. Environment,
*.
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x9RO1R

14: 38.
Pitchammal, V., Subramanian, G., Ramadevi, P. and Ramanathan, R. 2009.
The study of water quality at Madurai, Tamilnadu, India. Nature Envi-
ronment and Pollution Technology, 8(2): 355-358.
Regumathan, P., Beauman, W.H. and Kreusch, E.G. 1983. Efficiency of
point of use treatment devices. J. Am. Water Works Assoc., 75(1): 42.
Renn, C. E. 1970. Investigating Water Problems. LaMotte Chemical Prod-
ucts Company, Chestertown, Maryland. 50 pp.
Sallae, A.J. 1974. Waterborne diseses. In: Fundamental Principles of Bac-
teriology. Seventh Edition, Tata McGraw Hill Publishing Company
Ltd., New Delhi.
Sastry, K.V. and Rahee, P. 1988. Physico-chemical and microbiological
characteristics of water of village Kanneli, Haryana. Proc. Academy
of Environmental Biology, 7(1): 103-108.
Sawyer, C.N. and McCarty, P.L. 1967. Chemistry for Sanitary Engineers.
2nd edition. McGraw-Hill Series in Sanitary Science and Water Re-
sources Engineering, McGraw-Hill, Toronto.
Tihansky, D.P. 1974. Economic damages from residential use of mineral-
ized water supply. Water Resour. Res., 10(2): 145.
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Vol. 2, Health Criteria and Other Supporting Information, Geneva.


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Pavithra S. Reddy and N. Nandini
Department of Environmental Science, Bangalore University, Bangalore-560 056, Karnataka, India

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,1752'8&7,21
Solid waste means unwanted materials or substances that are
left or discarded after use, also included in it are by-prod-
ucts of process lines or materials that may be required by
law to be disposed off (Okecha 2000). Solid wastes can be
classified in a number of ways such as on the basis of source,
environmental risks, utility and physical property. On the
basis of source, which is commonly used, solid wastes are
classified as: municipal solid wastes, industrial solid wastes,
agricultural solid wastes, mining and mineral wastes, con-
struction and demolition wastes, healthcare wastes, radio-
active (nuclear) wastes, human and animal wastes. The gen-
eration of solid waste from household, industries, markets,
abattoir and shops result in improving the standard of living
of the inhabitants. The most common method of waste dis-
posal in India is dumping on land, because it is the cheapest
method of waste disposal. It, however, requires large area
and proper drainage. The land disposal of municipal and in-
dustrial solid waste is potential cause of groundwater con-
tamination. Uncontrolled dumping of waste is subjected to
leaching by percolating rain water and surface water or by
groundwater contact with landfill. These solid wastes can as
well contaminate groundwater. The generated leachate con-
tains high levels of BOD, COD, nitrates, chloride, alkalin-
ity, trace elements, hazardous wastes, etc. Leaching of water
from inappropriate dumping sites produce a foul smell and
is a nuisance to people living in surrounding areas
(Omofonmwan et al. 2009). The objective of the paper is
qualitative analysis of the leachate and the groundwater in
and around the land fill site of Mavallipura.
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Study area: Mavallipura landfill site is located about 20
km north of Bangalore city, 5.6 km away from the critical
defence zone close to Yelahanka air force base. It is about
2.5 km away from the flow of River Arkavathi. With the
tacit approval of Bhruhat Bangaluru Mahanagara Palike
(BBMP), every day since May 2003, about 200 truck loads
weighing about 2.5 to 3 tons of municipal solid waste each
from some of the northern wards of Bangalore, are being
dumped on Mavallipura landfill site. The site was handed
over to Ramky Infrastructure Ltd. in 2004 for composting.
Methodology: The utility of water is limited by its quality,
which may make it unsuitable for a particular purpose. There-
fore, assessment of water quality is an important aspect of
water evaluation and standard of living of the people. Solid
waste generated leachate and groundwater samples in and
around Mavallipura were collected and analysed for various
physico-chemical parameters like pH, electrical conductiv-
ity (EC), total dissolved solids (TDS), chemical oxygen de-
mand (COD), biochemical oxygen demand (BOD), total
hardness, calcium hardness, sulphates, sodium, potassium
and chlorides following standards methods (APHA 2005).
The results of leachate and groundwater samples were com-
pared with standards prescribed by WHO (1997), Bureau of
Indian Standards (BIS 1993) and Indian Council of Medical
Research (ICMR 1975) for disposal on surface waterbodies.
5(68/76$1'',6&866,21
Physico-chemical analysis of leachate samples: Physico-
chemical characteristics of the leachate depend primarily on
 Pavithra S. Reddy and Nandini N.

the waste composition and water content in the waste. The
characteristics of the leachate samples collected from the
Mavallipura landfill site are presented in Table 1. The In-
dian standards for composting and treated leachate call for a
complete cover of the waste storage area. If such storage area
is present in an open area, then it must be provided with
impermeable base with facility for collection of leachate and
surface water run-off into lined drains leading to leachate
treatment facility. Based on this notion, environmental analy-
sis to determine the extent of pollution caused by untreated
leachate into the groundwater was carried out. The average
pH was 8.37 (standards for leachate discharge into inland
surface water = 5.5-9.0). Electrical conductivity was above
the limit at 7937 micromhos/cm. Total dissolved solids
(TDS) was high at 4445 mg/L (permissible limit = 2100
mg/L). The relatively high values of EC and TDS indicate
the presence of inorganic materials in the samples. Chemi-
cal oxygen demand (COD) on an average was more than
twenty times as high at 5027 mg/L (disposal standards =
250 mg/L). Biochemical oxygen demand (BOD) was also
very high at an average of 3556 mg/L (permissible limit =
30 mg/L). The presence of high values of COD and BOD
indicate the organic strength. Total alkalinity was at 2505
mg/L. Total hardness of the leachate samples was above the
limit at an average 905 mg/L. Calcium hardness and magne-
sium hardness were at an average of 732 mg/L and 172
mg/L respectively (Table 1). The average sulphate content
was found to be 377 mg/L which was slightly higher than
the prescribed limit of 250 mg/L. An average chloride con-
tent in the samples was higher at 309 mg/L than the pre-
scribed limit of 250 mg/L. The nitrate content in the leachate
samples was 394 mg/L. An average sodium and potassium
contents in the leachate samples were in the range of 1205
mg/L and 855 mg/L respectively.
Physico-chemical analysis of groundwater samples: The
underground water of the study area is used for domestic
and other purposes. Table 2 shows the desirable and maxi-
mum permissible limit recommended by BIS (BIS 1993),
World Health Organization (WHO 1997) and Indian Coun-
cil of Medical Research (ICMR 1975). The pH of all the
groundwater samples was neutral, the range being 7.4 to 7.8.
The average pH of the groundwater samples was found to be
7.5. The EC is a valuable indicator of the amount of material
dissolved in water. The EC in the study area was found to be
high ranging between 610 and 977 µS/cm. The average EC
was 811 µS/cm. These high conductivity values obtained for
the underground water near the landfill are an indication of
its effect on the water quality. TDS indicates the general
nature of water quality or salinity. The range of TDS is be-
tween 389 mg/L and 789 mg/L with the average value of
637 mg/L. This high value of TDS may be due to the leach-
ing of various pollutants into the groundwater. The high
concentrations of TDS decrease the palatability and may
cause gastro-intestinal irritation in human and may also have
laxative effect particularly upon transits (WHO 1997). COD
is a measure of oxygen equivalent to the organic matter con-
tent of the water susceptible to oxidation by a strong chemi-
cal oxidant and thus is an index of organic pollution. The
COD level in the groundwater samples ranged from 243
mg/L to 436 mg/L with an average value of 331 mg/L indi-
cating the presence of organic contaminants in the water and
can be used as organic indicator to assess the groundwater

Table 1: Physico-chemical analysis of leachate samples.

Parameters Standards of disposal Sample 1 Sample 2 Sample 3 Sample 4

of treated leachate
(MPL by BIS and
ICMR for surface water)

pH 5.5-9.0 8.4 8.6 8.2 8.3

EC Not specified 8140 10420 7560 5630
COD 250 mg/L 2820 14420 1690 1180
BOD 30 mg/L 1566 10785 978 898
TDS 2100 mg/L 4060 7070 3910 2740
T. Alkalinity 600 mg/L 1800 4760 1670 1780
T. Hardness 600 mg/L 850 1200 760 810
Ca Hardness 500 mg/L 620 940 640 730
Mg Hardness 416 mg/L 230 260 120 80
Sodium (Na) 200 mg/L 920 2305 838 760
Potassium (K) - 1010 1140 330 941
Sulphates 250 mg/L 350 456 358 345
Chlorides 250 mg/L 280 365 298 295
Nitrates - 380 425 398 376

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 LEACHATE CHARACTERIZATION AND ASSESSMENT OF GROUNDWATER POLLUTION 

Table 2: Physico-chemical analysis of the groundwater.

Parameters Tolerance level prescribed Sample 1 Sample 2 Sample 3 Sample 4 Sample 5

by BIS, ICMR and WHO

pH 6.5-8.5 7.6 7.8 7.6 7.5 7.4

EC 250 -750 micromhos/cm 610 977 812 872 786
COD NIL 243 436 327 323 327
BOD NIL 8.7 9.0 8.4 8.3 8.6
TDS 500 mg/L 389 789 606 716 685
T. Alkalinity 200 mg/L as CaCO3 174 124 132 120 144
T. Hardness 300 mg/L as CaCO3 283 396 314 264 278
Ca Hardness 187.5 mg/L as CaCO3 68 120 93 68 75
Mg Hardness 125 mg/L as CaCO3 215 276 221 196 203
Chlorides 250mg/L as Cl 98 146 125 124 131
Sulphates 250mg/L as SO4 94 156 136 128 136
Nitrates 45mg/L as NO3 4.2 6.5 3.8 2.4 3.1
Sodium 200 mg/L 34 49 39 32 29
Potassium 55 mg/L 1.2 1.6 1.4 1.3 1.2

Note: For EC, the standards are adopted from Manivasakam (2003)

pollution caused by landfill. Total alkalinity of the
groundwater samples was within the permissible limit with
the average value of 138 mg/L. The high alkalinity imparts
an unpleasant taste, and may be deleterious to human health
with high pH, TDS and total hardness. Multivalent cations,
particularly Mg2+ and Ca2+ are often present at a significant
concentration in natural waters. These ions are easily pre-
cipitated and in particular react with soap to make it diffi-
cult to remove scum. Total hardness is normally expressed
as the total concentration of Ca2+ and Mg2+ in mg/L equiva-
lent CaCO3. Total hardness ranged from 264 mg/L to 396
mg/L and the average value was 307 mg/L. Calcium hard-
ness and magnesium hardness ranged from 68 mg/L to 120
mg/L and 196 mg/L to 276 mg/L respectively. Magnesium
salts are cathartic and diuretic and high concentration may
cause laxative effect. Chloride content in the water samples
ranged from 98 mg/L to 146 mg/L. High chloride content of
groundwater is likely to originate from pollution sources such
as domestic effluents, fertilizers, septic tanks and from natu-
ral sources such as rainfall and the dissolution of fluid in-
clusions. Increase in chloride level is injurious to people suf-
fering from diseases of heart or kidney (WHO 1997). Sul-
phate in the water samples ranged from 94 mg/L to 146
mg/L. Nitrates ranged from 2 mg/L to 6 mg/L. The average
value of sodium and potassium content was 37 mg/L and
1.3 mg/L respectively. The high concentration of sodium
may pose a risk to persons suffering from cardiac, renal and
circulatory diseases.
&21&/86,216
The moderately high concentration of EC, TDS, Cl-, SO2-4,
NO-3, Na+, etc. in groundwater near landfill deteriorates its
quality for drinking and other domestic purposes. Further,
the presence of Cl-, NO-3 and COD can be used as tracer with
relation to leachate percolation. From the groundwater moni-
toring it is clearly evident that the leachate generated from
the landfill site is affecting the groundwater quality in the
adjacent areas through percolation in the subsoil. As there is
no natural or other possible reason for high concentration of
these pollutants, it can be concluded that leachate has sig-
nificant impact on groundwater quality near the area of
Mavallipura landfill site. The groundwater quality improves
with the increase in depth and distance of the well from the
pollution source. Although, the concentrations of few con-
taminants do not exceed drinking water standard even then
the groundwater quality represent a significant threat to pub-
lic health. This will go a long way to protecting natural re-
sources such as water that are degraded by these solid wastes.
Solid waste handling, controlling and monitoring techniques
in Mavallipura must be geared towards achieving quality
environmental condition for man to live in.
5()(5(1&(6
Anandhaparameshwari, N., Hemalatha, S., Vidhyalakshmi, G.S. and
Shakunthala, K. 2007. Groundwater quality characteristics at
Sivalingampillai layout, Udumalpet, Tamilnadu. Nature Environment
and Pollution Technology, 6(2): 333-334.
APHA 2005. Standard Methods for the Examination of Water and
Wastewater. American Public Health Association, Washington, DC.
Chandrasekar, P. and Ayyappan, S. 2006. Impact of municipal solid waste
(MSW) dumping on groundwater quality - A Case study. Poll Res.,
25(1): 31-34.
ICMR 1975. Manual of Standards of Quality of Drinking Water Supplies.
Indian Council of Medical Research, New Delhi.

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Pavithra S. Reddy and Nandini N.
BSI 1993. Indian Standard for Drinking Water Specification, IS: 10500
1991 and Amendment, Bureau of Indian Standards, New Delhi.
Okecha, S. A. 2000. Pollution and Conservation of Nigeria Environment.
T’ Afrique International Associates, Oweri, Nigeria.
Omofonmwan, S. I. and Eseigbe, J. O. 2009. Effects of solid waste on the
quality of underground water in Benin Metropolis, Nigeria. J. Hum.
Ecol., 26(2): 99-105.
Ramadevi, P., Subramanian, G., Pitchammal, V. and Ramanathan, R. 2009.
The study of water quality of Ponnaravathy in Pudukkottai district,
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
Tamilnadu. Nat. Environ. & Poll. Technol., 8(1): 91-94.
Ramesh, N., Meenamabal, T. and Murugan, K. 2009. Quantification, char-
acterization and leachate analysis of the municipal solid waste from
Erode Municipality, Tamilnadu, India. Nature Environment and Pol-
lution Technology, 8(1): 21-28.
WHO 1997. International Drinking Water Supply and Sanitation Decade-
Review of National Baseline Data, WHO Offset Publication, No. 85,
World Health Organization, Geneva.
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Harminder Kaur, Radhe Shyam and R. Amutha
Department of Applied Sciences, PEC University of Technology, Chandigarh-160 012, India

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,1752'8&7,21
Effective methods available for wastewater treatment gen-
erally involve costlier adsorbents and hence not economi-
cally viable. Therefore, continuous research is going on to
develop newer low cost adsorbents from agricultural waste
(Sivaraj et al. 2002), domestic waste (Hameed 2009), indus-
trial waste (Wang et al. 2005), forest trees (Babu & Gupta
2008) and many more sources. Lignin content and or other
functional groups present on the surface of these adsorbents
are responsible for the uptake of metals and non metals from
aqueous solution. In India, it is popularly known as ‘Sau-
sage tree’ and locally known as ‘Balam khira’ This tree can
grow up to 20 m tall and widely planted for its ornamental
flowers, firewood and shade. A full grown tree bears hun-
dreds of fruits hanging in and each cylindrical fruit weighs
about 2.5 kg. Bark and fruits of this tree have demonstrated
medicinal properties (Owolobi et al. 2007, Olalye & Rocha
2007), but their utilization in India is negligible. Thousands
of Kigelia africana trees grow in the forest belts of Punjab,
Haryana and Himachal Pradesh. Fruits and leaf litter are
thrown into wasteland as there is no known use in this re-
gion. Finding an alternate method to convert this readily
available waste material into a valuable resource will cer-
tainly benefit the mankind. A variety of adsorbents were
developed from leaves (Kumar et al. 2006), stem (Anirudhan
et al. 2005), seeds (Gonzalez & Montoya 2009), fruits
(Inbaraj & Sulochana 2004) and almost all parts of many
plants and trees. However, no work has been reported on
this popularly grown Kigelia africana tree fruits or leaves
as a precursor to develop adsorbents, and therefore, the
present work has been undertaken.
0$7(5,$/6$1'0(7+2'6
Plant material: Kigelia africana belongs to Bignoniaceae
family. Mature ripe fruits were collected from Punjab Engi-
neering College campus, Chandigarh on 5th February, 2010.
They were washed with water, cut into small pieces and air
dried in the open for a few days and later further dried in an
electric oven at 110°C for 72 h and powdered in a mechani-
cal mill. 0.25 L of 1:1 conc. H2SO4 was added with 100g of
plant material powder kept in a 0.5 L beaker. This mixture of
fruit powder and sulphuric acid was heated in an electric oven
at 110°C for 24 hours. It was stirred once in an hour. The
slurry was washed thoroughly with water and finally with
distilled water and dried. Dry carbonized Kigelia africana
fruit powder (KAFP) material was stored in an airtight con-
tainer for further use.
Chemical reagents: All chemicals used in this study were
of analytical grade and a stock solution of 1000 mg/L was
prepared by dissolving 3.801g of Cu(NO3)2.3H2O in 1 L of
deionized water. Working standards were obtained by dilut-
ing stock solution with appropriate volume of deionized
water. 0.05 N NaOH and 0.05 N HNO3 were used to main-
tain pH of initial metal ion solutions. Conc. H2SO4 was em-
ployed in the carbonization of the plant material.
Batch adsorption studies: A series of 0.1 L of metal ion
solutions in the concentration range of 10 mg/L to 25 mg/L
were taken in 0.25 L conical flasks. 0.1 g of KAFP adsorbent
material was added to each conical flask solution and they
were closed with screw corks. The suspensions were shaken
in a water bath cum shaker for 2 h and thereafter, centrifuged.
The clear supernatant solution was analysed in AAS (ECIL,
 Harminder Kaur et al.

India) for determination of copper concentration. Table 1: Langmuir equilibrium parameter and nature of the isotherm
applied.
Contact time studies: 0.1 L of metal ion solution (20mg/L)
was taken in 5 different 0.25 L conical flasks. 0.1 g of KAFP Value of RL Nature of isotherm applied
adsorbent was added to all conical flask solutions and they
RL > 1.0 Unfavourable
were shaken for 15, 30, 60, 90 and 120 minutes respectively. RL = 0 Linear
Clear solution was drawn from these conical flasks after shak- RL < 1.0 Favourable
ing and analysed for copper. RL = 0 irreversible
Adsorbent dose studies: 0.1, 0.15, 0.2 and 0.25g of KAFP
Table 2: Langmuir constants for the adsorption of Cu(II) ions on KAFP
was added to four 0.25 L conical flasks containing 0.1 L of from aqueous solution.
20 mg/L metal ion solution. All flasks were closed with screw
corks and shaken in a laboratory shaker for 2 h. Clear solu- Adsorbent Sorption capacity Reference
(Q)(mg/L)
tions were drawn after 30 min and analysed for copper con-
centration. KAFP 21.74 Present work
pH studies: Batch adsorption experiments were conducted Sunflower stalk 29.3 Sun and Shi (1998)
Saw dust 13.6 Ajmal et al. (1998)
to find adsorption of copper (II) ions onto KAFP at different Palm oil fibre 19.8 Low et al. (1993)
initial metal ion concentrations. pH of the solutions were Sago waste 12.42 Quek et al. (1998)
maintained by adding either NaOH or HNO3.
Table 3: Equilibrium parameter values for the adsorption of Cu(II) on KAFP
5(68/76$1'',6&866,21 from aqueous solution.

Batch adsorption data obtained at different initial metal ion Ce (mg/L) Copper(II)
concentrations ranging from 10 mg/L to 25 mg/L fit well
10 0.556
into Langmuir adsorption isotherm equation. Langmuir con-
15 0.455
stants Q and b are calculated from slope and intercept of 20 0.385
linear Langmuir plot for the adsorption of Cu(II) ions on 25 0.334
KAFP from aqueous solution (Fig. 1). Following Langmuir
equation was used. Table 4: Effect of adsorbent amount on Cu(II) ions uptake by KAFP from
aqueous solution.

% Cu(II) Amount of Amount of Cu(II)

Where, Ce = equilibrium concentration, qe = amount removal adsorbent (mg) adsorbed (mg/g)
adsorbed on to the adsorbent (mg/g), Q = sorption capacity 87.25 100 21.3
(mg/g), b = sorption energy (L/mg). 90.35 150 19.2
Another dimensionless equilibrium parameter, RL was 89.3 200 17.9
89.85 250 15.7
calculated from the following relation.

RL value gives the information regarding the nature of

isotherm applied (Table 1).
Langmuir constant, Q values for Cu(II) adsorption on
KAFP and other low cost adsorbents are given in Table 2. RL
values between 0 and 1 confirm applicability of Langmuir
isotherm equation for the adsorption of Cu(II) from aque-
ous solution. Langmuir equilibrium parameter at different
initial metal ion concentrations for Cu(II) adsorption on
KAFP is presented in Table 3. Percent removal of Cu(II)
ions decreased from 90.9 to 70.16 when initial metal ion
concentration was increased from 10 mg/L to 25 mg/L.
Whereas, metal ion uptake by KAFP increased from 4.56
mg/g to 8.77 mg/g. Freundlich isotherm is shown in Fig. 2. Fig. 1: Langmuir plot for the adsorption of Cu(II) ions on
Though R2 value is very close to 1 (0.966), 1/n and Kf values KAFP from aqueous solution.

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 KIGELIA AFRICANA FRUIT CARBON AS AN ADSORBENT FOR REMOVAL OF COPPER(II) 

Fig. 2: Freundlich plot for the adsorption of Cu(II) ions on
KAFP from aqueous solution.
confirm the applicability of Langmuir isotherm equation in
this case. Contact time studies revealed that equilibrium was
achieved well within 30 minutes (Fig. 3). Percent removal
of Cu(II) ions increased from 20 to 92 approximately on in-
creasing adsorbent dose from 0.1 g to 0.25 g but amount of
metal ion intake by the adsorbent decreased from 21.3 mg/g
to 15.7 mg/g. This is due to the increase in the available
adsorbent surface (Table 4). 0.05 N HNO3 and 0.05 N NaOH
solutions were used to adjust pH of initial metal solutions.
Effect of pH on copper adsorption onto KAFP was studied
at different pH values of 2-8. Uptake of metal ion was mini-
mum at lower pH (2-3) and increases at higher pH values (3-
7).
Mechanism of adsorption: FTIR peaks at 2365.5cm-1
(-C=C-stretching), 1377.9cm-1and 1583.7cm-1 (carboxylate
ion), 2923cm-1 (methylene-C-H stretching) and 3368.5cm-1
(intermolecular hydrogen bond) were observed. Ion ex-
change may be the most predominant mechanism as acidic
carboxylate groups are found on the surface of the adsorb-
ent sample. At much lower pH, H+ ions compete with metal
cations resulting in much less metal removal from the aque-
ous solution. But at higher pH values, increase in metal up-
take is due to the decrease in H+ ions in the solution.
&21&/86,216
• Carbonization of KAFP was achieved by heating with
conc. H2SO4 at 110°C for 24 h.
• Adsorption data obey Langmuir adsorption isotherm equa-
tion.
• Equilibrium was achieved within 30 minutes.
• The study reveals the effectiveness of Kigelia africana
fruit powder carbon in removing copper from aqueous
solution as it showed comparable adsorption capacity
value with other known low cost adsorbents.
• Maximum adsorption capacity of KAFP was 21.74
mg/g.
• 90.7% Cu(II) removal was achieved when 0.1 g of KAFP
added to 0.1 L of 10 mg/L solution.
• Presence of acidic functional groups like carboxylate ions
on the surface of the adsorbent was confirmed by FTIR
spectrum.

Fig. 3: Effect of contact time on Cu(II) adsorption on KAPF. Contact time revealed that equilibrium
was achieved well within 30 minutes.

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 Harminder Kaur et al.
• Ion exchange may be one of the possible mechanisms of
adsorption in this case.
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Babu, B.V. and Gupta, S. 2008. Adsorption of Cr (VI) using activated neem
leaves: Kinetic studies. Adsorption, 14(1): 85-92.
Gang Sun and Weixing Shi 1998. Sunflower stalks as adsorbents for the
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1324-1328.
Hameed, B. H. 2009. Spent tea leaves: A new non-conventional and low-
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L. P. Rema*
Division of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science &
Technology, Kochi, Kerala, India
*Current Address: P. G. and Research Department of Zoology, Maharaja’s College, Ernakulam, Kerala, India

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Some structural peculiarities of the lysosomal membrane,
which is not yet clear, imparts the fundamental property of
structure linked latency of lysosomal enzymes. This stabil-
ity is essential for the normal functioning of the cell and it
protects rest of the cell from the destructive effects of the
enzymes. Alteration in the membrane causes release of en-
zymes and is of outstanding pharmacological significance
(Woessner Jr. 1971, Kocchar & Larsson 1987).
Lysosomes are one of the most fascinating targets of
heavy metals. Investigations of Sternlieb & Goldfischer
(1976) have demonstrated that lysosomes are the most im-
portant sites of metal compartmentation in the cell. Their
capacity to store and sequester a wide range of metal ions
has been acknowledged by Allison (1969). An activation of
lysosomal system involving an increase in the number and
size of lysosomes was reported by various researchers (Myers
et al. 1987, Studnicka 1983, Daoust et al. 1984, Weis et al.
1986, Sauer & Watabe 1989). This step clearly represents an
adaptive and protective response to injury.
Heavy metals, which accumulate in lysosomes, stimu-
late the lipid peroxidation process and at the same time, in-
hibit the native defence mechanisms involved in prevention
of lipid peroxidation. This results in formation of lipofuscin
granules within the lysosomes. Lipofuscin can complex
heavy metals and is a cardinal mechanism in heavy metal
homeostasis (Viarengo 1985). Lipofuscin can later get trans-
formed into an insoluble polymer that includes part of the
bound metals which then become unavailable to the cell.
Thus, though the metals within the lysosomes enhance lyso-
somal lipid peroxidation and alter the normal physiology of
these organelles, they augment the amount of lipofuscin gran-
ules that can trap toxic metals in relatively stable form.
Though lysosomes are noted for their sequestration and
accumulation of metals and various other chemicals, many
of these substances are capable of destabilizing the lysosomal
membrane, if the storage capacity is surpassed. Activation
and leakage of the previously latent degradative lysosomal
enzymes, which have got a high potential for the catabolic
disruption of cellular systems, ensue and is of considerable
environmental consequence (Bayne et al. 1978). These lyso-
somal membrane changes are perhaps, the earliest detect-
able alterations of cellular damage and hence, sensitivity of
lysosomes to environmental pollutants including heavy met-
als ranks lysosomal responses as early warning systems for
detection of the disturbances in the surroundings.
The present study attempts to extend the lysosomal en-
zyme releases assay technique to Oreochromis mossambicus
exposed to two different heavy metals. This will, in turn,
helps to assess its effectiveness as a sensitive index of heavy
metal stress and its applicability as a biochemical warning
of environmental alterations.
0$7(5,$/6$1'0(7+2'6
Male Oreochromis mossambicus of average length 10±1 cm
were collected from Rice Research Institute, ICAR, Kochi.
 L .P. Rema
They were brought to the laboratory and acclimated for one
month. The fish were fed daily and water was changed after
consumption of the supplied food.
Feeding of the fish was suspended 24 hour prior to the
experiment. Fish were stocked as three groups at a density
of one fish per 5 litre. One group served as the control and
contained dechlorinated tap water only and the others con-
tained 1/10th of 96 hour LC50 of zinc (0.42 ppm) and mer-
cury (0.1 ppm) respectively. The medium was renewed daily
with minimum disturbance for the fish. After two days of
exposure to the toxicant, fish were caught, liver was excised,
water blotted off and weighed accurately. The process was
repeated after 4, 6 and 8 days of exposure. Samples were
homogenized in ice-cold isotonic sucrose solution contain-
ing 2mM mercapto-ethanol. Stability of lysosomal mem-
brane was determined following the procedures of Philip &
Kurup (1977, 1978) and Rao & Sisodia (1986). Acid phos-
phatase, the marker enzyme of lysosomes, released from the
organelle was assayed following the procedure of Anon
(1963). To the enzyme extract, 0.01 mM EDTA (Verity &
Reith 1967) was also added to chelate any heavy metal
present so as to nullify the chance of direct inhibition of
enzyme by the metal.
Similar procedure was followed for the in vitro studies
of the lysosomal latency, where the lysosomal rich fraction
obtained from liver of acclimated fish was suspended in iso-
tonic sucrose solution containing mercapto-ethanol and was
incubated at 2 and 20 ppm of zinc and mercury solutions.
5(68/76
The effect of zinc and mercury on the stability of lysosomal
membranes is shown in Table 1. Effect of in vitro exposure
to zinc and mercury on the cell free preparations of lysosomes
is presented in Table 2. The lysosomal lability indices, ob-
tained as a function of time and function of various
xenobiotics, were analysed using ANOVA technique. Wher-
ever, the effects were found to be significant, least signifi-
cant difference (LSD) at 5% level was calculated.
',6&866,21
Lysosomes play a crucial role in the isolation and sequestra-
tion of heavy metals and thus, relieve the biological machin-
ery from the toxic effects of heavy metals. But any excess of
the heavy metal, beyond its storage capacity, causes leakage
of degradative enzymes and derangement of cell functions.
Change in the membrane stability of lysosomes is one of
the first alterations due to metal exposure and bears a quan-
titative relationship to the magnitude of stress response
(Bayne et al. 1979, 1982, Cajaraville et al. 2000, Camus et
al. 2000, Marchi et al. 2004). Thus, lysosomal membrane
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
stability is a highly sensitive measure of the functional state
of the cell and provides an ideal starting point for probing
into the generalized cellular deterioration.
Presence of functional groups like thiol and disulphides
in the membrane plays a key role in the integrity and perme-
ability properties of the membrane and also in the structure
linked latency. Mercuric compounds irreversibly damage the
limiting membrane with the formation of mercaptide bridges.
Moreover, the lipid peroxidation ensuing for the production
of lipofuscin granules to bind mercury for homeostasis, also
leave an unfavourable alteration in the physiology of lyso-
somal lamina. Mercury also suppresses the natural mecha-
nism which inhibits lipid peroxidation.
Heavy metals have an “all or none” effect on individual
lysosomes (Verity & Reith 1967). The values of acid phos-
phatase released, represent the proportion of the lysosomes
whose threshold has been surpassed. The differences in in-
dividual lysosomal responses can be attributed to morpho-
logical and biochemical parameters that may decide the rela-
tive availability of thiol groups to the metal ions in the
system.
The results of the present study indicates that as with
other detoxification systems, the accumulation and seques-
tration of mercury by lysosomes also is effective only until
the storage capacity of the lysosomes is overloaded or the
lysosomes are damaged directly by the accumulated contami-
nant as reported by Moore et al. (1985). The lysosomal mem-
brane is showing an initial stabilization and this is followed
by a progressive labilization. This indicates that sequestra-
tion of mercury by lysosomes, which relieves the biochemi-
cal machinery from their toxic effects, is efficient only up to
a limit. Thereafter, this detoxifying machinery becomes in-
effective resulting in the decline of the structure linked la-
tency of lysosomal enzymes.
On exposure to zinc, an initial labilization is seen prob-
ably due to the delayed onset of operation of the detoxifying
mechanisms against this essential metal. The onset of opera-
tion of detoxifying mechanisms for this essential metal is
not as fast as that for the toxic nonessential mercury. Results
of the in vitro studies with 2 and 20 ppm solutions of zinc
substantiate this hypothesis. A direct effect of metals on
membrane rather than on the lysosomal enzyme is possible
since all the metals remaining, which can cause any differ-
ence in activity of acid phosphatase, whose rate of release is
taken as the index of the lability of lysosomes, are chelated
and made unavailable to the system. Continued exposure to
zinc stabilizes the membrane either by binding to the struc-
tural components or by inhibiting metal catalysed lipid
peroxidation. Such a direct effect on membrane, suggesting
that metals, which form redox systems, catalyse the lipid
 EFFECT OF ZINC AND MERCURY ON LYSOSOMAL MEMBRANE STABILITY OF FISH 

Table 1: In vivo effect of Zn and Hg on stability of lysosomal membrane of O. mossambicus as a function of period of exposure. Stability of lysosomes is
assessed by following the activity of acid phosphatase (millimoles of p-nitrophenol formed/hour/g protein) released as a function of time at 37°C.

Time Control Mercury Zinc

(min.) Activity LLI Activity LLI Activity LLI

A. Two Days
00 02.96 ± 0.36 16.99 04.23 ± 0.26 15.68 03.52 ± 0.32 16.36
10 03.84 ± 0.18 22.07 05.27 ± 1.95 19.53 05.32 ± 0.20 24.76
20 03.87 ± 0.26 22.24 05.50 ± 0.23 20.38 06.04 ± 0.17 28.11
30 04.22 ± 0.11 24.26 05.73 ± 0.17 21.25 06.59 ± 0.21 30.67
B. Four Days
00 08.13 ± 0.91 34.80 11.00 ± 2.17 35.60 03.89 ± 1.11 19.13
10 08.40 ± 0.33 35.95 10.93 ± 0.88 35.12 05.39 ± 0.38 26.45
20 08.67 ± 0.37 37.12 12.80 ± 0.44 41.13 05.44 ± 0.17 26.72
30 09.00 ± 0.29 38.53 12.82 ± 0.34 41.22 06.21 ± 0.15 30.48
C. Six Days
00 09.28 ± 1.05 22.60 15.71 ± 0.84 21.18 04.42 ± 0.26 17.87
10 09.55 ± 0.29 23.25 19.37 ± 0.85 26.12 05.58 ± 0.30 22.58
20 10.47 ± 0.45 25.50 24.51 ± 1.75 33.04 05.53 ± 0.46 22.37
30 09.84 ± 0.74 23.97 34.67 ± 1.20 46.74 05.52 ± 0.07 22.38
D. Eight Days
00 02.53 ± 0.20 13.45 02.94 ± 0.57 09.77 02.29 ± 0.57 10.31
10 04.06 ± 0.39 21.56 09.71 ± 0.23 32.29 04.17 ± 0.31 18.83
20 04.70 ± 0.38 24.95 09.57 ± 0.29 31.85 04.81 ± 0.17 21.68
30 05.63 ± 0.49 29.92 09.61 ± 0.50 31.96 04.98 ± 0.45 22.47

Two days - (Between groups, p<0.01, LSD at 5% level 0.870; Between time, p<0.01, LSD at 5% level 0.754)
Four days - (Between groups, p<0.01, LSD at 5% level 1.038; Between time, p<0.01, LSD at 5% level 0.899)
Six days - (Between groups, p<0.01, LSD at 5% level 9.18; Between time, not significant 5% level)
Eight days - (Between groups, p<0.001, LSD at 5% level 2.62; Between time, p<0.05, LSD at 5% level 2.27)
Lysosomal lability index (LLI) is the activity of acid phosphatase released expressed as percentage of the total activity of lysosomal lability acid
phosphatase. Values are the mean of six different experiments ± SD.

Table 2: In vitro effect of different concentrations of Zn and Hg on the stability of lysosomal membrane of O. mossambicus. Stability of lysosomes is
assessed by following the activity of acid phosphatase (millimoles of p-nitrophenol formed/hour/g protein) released as a function of time at 37°C.

Time Control Mercury Zinc

(min.) Activity LLI Activity LLI Activity LLI

A. 2 ppm
00 01.33 ± 0.13 12.54 01.58 ± 0.11 14.89 00.61 ± 0.06 05.80
10 01.99 ± 0.10 18.74 02.11 ±0.12 19.94 03.03 ± 0.56 28.59
20 02.34 ± 0.10 22.04 02.45 ± 0.07 23.12 03.74 ± 0.57 35.29
30 02.96 ± 0.15 27.92 02.53 ± 0.07 23.87 04.83 ± 0.25 45.62
B. 20 ppm
00 01.64 ± 0.11 06.70 01.20 ± 0.19 04.91 00.23 ± 0.04 00.93
10 02.15 ± 0.10 08.77 01.80 ±0.13 07.34 00.61 ± 0.17 02.48
20 02.75 ± 0.14 11.23 02.63 ± 0.13 10.71 00.61 ± 0.06 02.50
30 03.30 ± 0.15 13.48 02.53 ± 0.35 10.34 00.61 ± 0.07 02.48

At 2 ppm - (Between groups, Not significant; Between time, Not significant)

At 20 ppm - (Between groups, p<0.001, LSD at 5% level 0.69; Between time, p<0.05, LSD at 5% level 0.599)
Lysosomal lability index (LLI) is the activity of acid phosphatase released expressed as percentage of the total activity of lysosomal lability acid
phosphatase. Values are the mean of six different experiments ± SD.
peroxidation and cause membrane damage has also been re- mollusks (Sadikaji et al. 2010). Ability of zinc to stabilize
ported by Chvapil (1973). Lysosomal enzyme release assay lysosomal membrane has been demonstrated by Sternleib &
(LERA) thus is effective as a sensitive index of heavy metal Goldfischer (1976) also.
stress and can be applied as a biochemical warning of envi- To sum up LERA expressed in terms of lysosomal lability
ronmental alterations. LERA has been substantiated as a sen- index (LLI) clearly reflects breakdown of the adaptive ca-
sitive indicator of numerous environmental stresses in pacity of fish to toxic injury. It is to be realized that lability

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 L .P. Rema
of lysosomal membrane is not indicative of any specific
stress, but many factors can cause the same response and so
is a general index of stress. However, LERA is a very good
indicator to monitor the effects of environmental pollutants
on biomembranes.
5()(5(1&(6
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Thermal Engineering Group, Central Mechanical Engineering Research Institute, Durgapur-713 209, W. B., India

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The rapid increase in the manufacturing, storage and han-
dling of a wide variety of hazardous chemicals have brought
about a situation in India where the safety of workers em-
ployed in the factory and the general public living in the
vicinity of the factory could be endangered at any point of
time because an accident involving these chemicals may
cause injury or loss of life or damage property including ad-
verse effect on environment or disruption in and around the
factory. Bhopal and other similar catastrophes gave rise to a
new classification of industries having potential for major
accidents such as major emission, fire or explosion with off-
site consequences on human life and environment. The
Bhopal incident triggered interest from industry, academia
and legislators and is widely acknowledged as one of the
defining events in the history of process-safety. In spite of
the improved design procedures being followed by the in-
dustry to set up the plants, there are accidents of various de-
grees of severity taking place in chemical industry. Most of
them are minor in nature and get analysed by the concerned
industry itself. But, some of them turn out to be of very seri-
ous nature and result in death and damage not only inside
the factory premises but also outside.
As per the Factories Act, carbon-monoxide poisoning is
one of the noticeable diseases throughout the world. Coke-
oven and blast furnace gas consist of carbon-monoxide gas
and hence it is also very important from the medico-legal
aspect.
Under the Manufacture, Storage and Import of Hazard-
ous Chemicals Rules, 1989, amended in 2000, issued by the
Govt. of India, threshold quantities of hazardous substances
presenting major accidents are mentioned. The rules defined
hazardous chemicals based on their inherent characteristics
of toxicity, flammability and explosivity.
In India a total of 586 Maximum Accident Hazardous
(MAH) units and 75 hazardous chemicals are distributed
throughout the country. The distribution of MAH units and
hazardous chemicals across the various States of India
(Raghvan & Swaminathan 1996) is given in Table 1.
In Durgapur, one of the scenario identified is jet fire/
flare of coke-oven gas from coal gas plant and blast furnace
gas from steel plant. Hazards of jet fire/flare are illustrated
as an emergency scenario.
The study area and hazards of coke-oven and blast
furnace gas: Taking situation of Durgapur area in Burdwan
district located at 23.48° N and 87.32° E at 160 km from
Kolkata, there are 11 MAH installations as per “The Manu-
facture Storage and Import of Hazardous Chemicals Rules
(MSI)”. The MSI Rules enacted in 1989 under the Environ-
ment Protection Act, 1986, subsequently amended in 1994 by
Govt. of India represent a comprehensive attempt to regu-
late major accident hazard installations, comprising facto-
ries, isolated storage and cross-country pipelines. All these
industries have their own on-site emergency plan. In this
context, the sub-division is also having off-site emergency
plan for which District Collector of Burdwan acts as Emer-
gency Authority. Coke-oven gas is categorized as a hazard-
ous/toxic chemical, containing mainly hydrogen and methane.
On the other hand blast furnace gas is also categorized as a haz-
ardous/toxic chemical, containing mainly carbon monoxide.
 Biswajit Ruj
Coke oven gas, a hazardous gas, also called coal gas, is
the gas obtained when coal is heated in an air tight place in
pyrolysis process. It consists mainly hydrogen and meth-
ane. Coke oven gas is burned to produce heat in industrial
plants and in homes. The coke-oven gas contains hydrogen-
55%, methane-24%, ammonia-3.5%, carbon monoxide-9%,
carbon dioxide-3%, nitrogen-3%, oxygen-0.5%, and hydro-
carbons and others (ethane, ethylene, benzene, cyclopenta-
diene, toluene, naphthalene, hydrogen sulfide, water vapour,
hydrogen cyanide, cyanogen, nitric oxide)-2% . Coke-oven
gas is yellowish in colour with pungent odour and lighter
than air (0.45 times lighter).
Blast furnace gas is a by-product of blast furnaces, also a
hazardous gas that is generated when the iron ore is reduced
with coke to metallic iron. It has a very low heating value,
about 93 BTU/cubic foot, because it consists of about 56%
nitrogen,16.5% carbon dioxide, which are not flammable.
Hydrogen 2% and methane 0.5 % are also present in this
gas. The rest 25% is carbon monoxide, which has a fairly
low heating value. It is commonly used as a fuel within the
steel works, but it can be used in boilers and power plants
equipped to burn it. It may be combined with natural gas or
coke oven gas before combustion or a flame support with
richer gas or oil is provided to sustain combustion. Blast
furnace gas is sometimes flared without generating heat or
electricity.
Blast furnace gas is generated at higher pressure and at
about 100-150°C in a modern blast furnace. Auto-ignition
point of blast furnace gas is approx. 630-650°C and it has
LEL (Lower Explosive Limit) of 27% and UEL (Upper Ex-
plosive Limit) of 75% in an air-gas mixture at normal tem-



perature and pressure. Blast furnace gas is almost colourless
(mild whitish), odourless and heavier than air. Higher con-
centration of carbon-monoxide makes the gas hazardous.
A jet fire/flare is a pressurized stream of combustible gas
or atomized liquid that is burning. If such a release is ignited
soon after (i.e, within 2-3 minutes), the result is an intense
jet fire/flare. This jet fire stabilizes to a point, which is close
to the source of release, until the release is stopped.
The consequences arising out of the release of flamma-
ble chemicals and toxins are dependent on the prevailing
meteorological conditions. The meteorological data of
Durgapur with respect to wind speed and wind direction are
given in Table 2. The average values show that there are
predominantly two directions from which wind blows in this
area, viz., south and north.
-
 
.


- /
Consequences of hazards of coke-oven gas and blast
furnace gas: Prediction and consequence assessment of
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eventual accidents are not easy to accomplish due to the un-
certainty of the kind and the amount of hazardous materials
present at a certain time and place (ILO 1990). Neverthe-
less, typical scenarios of possible accidents can be devel-
oped considering the maximum quantities of typical dan-
gerous substances often arriving at the port. Quantification
of risk and consequences assessment associated with acci-
dents involving hazardous materials have been carried out
by several authors (Ruj et al. 2006, Ruj et al. 2009, Spadoni
& Uguccioni 2003, Tixier et al. 2002, Ditali et al. 2000, Khan
& Abbasi 1999, Ruj 2005, Ruj & Chatterjee 2007).
Considering all the possible identifications with regards
to hazards of coke-oven and blast furnace gas, the following
hazardous scenarios at Durgapur on the basis of engineering
judgement and expertise in the field of risk analysis were
studied.
Release of toxic/flammable gases from storage tanks stor-
ing coke-oven gas and blast furnace gas: The maximum
credible loss scenarios have been assessed for possible inad-
vertent accidents by applying Complex Hazardous Air Re-
lease Model (CHARM) software package. CHARM is a mod-
elling program that calculates and predicts the dispersion
and concentration of airborne plumes from released chemi-
cals. CHARM also predicts the footprints of thermal radia-
tion and overpressure.
In this work, accidental scenario based on real data is
presented. The results show that an eventual accident will
extensively damage the surroundings, even causing signifi-
cant fatalities in the affected population.
 .
Jet fire/flare from major leakage of coke-oven gas in coal
gas storage tank: About 56,000 cu. m coke-oven gas is
stored in one gas holder at an approximate pressure of 350
mm water gauge. The assumptions of this scenario are heavy
leakage of gas due to rupture of outgoing/incoming pipe due
to sabotage, riot, corrosion, etc., and getting ignited from an
external source.
The thermal consequence models provided a set of radi-
ant heat fluxes expressed in kW/m2 at a set of distances at
which the following heat flux thresholds occurred: 4.7
kW/m2, a pain threshold, 12.5 kW/m2, first-degree burn
threshold and 37.5 kW/m2, mortality threshold (AICE 1989).
It has been estimated by applying CHARM software that
damage distance for heat flux of 37.5 kW/m2 will have ap-
proximately 350 m radius and for heat flux of 4 kW/m2, it
will be about approximately 1 km. The total time duration
would be about 1.2 hours. Hence, serious damage will occur
to process equipments within 350 m radius zone. This will
also have disastrous effect on the work force in that area.
 HAZARDS OF COKE-OVEN GAS AND BLAST FURNACE GAS 

Table 1: State-wise distribution of major hazardous units (MAH) and haz- Table 2: Meteorological data.
ardous chemicals.
Month Wind speed Wind direction (deg.)
State MAH units Hazardous chemicals (m/s), Avg.
Min. Max.
Andhra Pradesh 35 24
Assam 7 10 May 0.79 182.11 2.59
Bihar 12 11 June 0.84 1.54 269.96
Delhi 19 8 July 1.12 3.55 0
Goa 8 9 Aug 1.20 0 51.04
Gujarat 112 32 Sept 5.92 0 310.66
Haryana 7 4 Oct 8.61 0 45.81
Jammu & Kashmir 7 4 Nov 6.33 238.0 44.43
Karnataka 26 14 Dec 1.58 282.04 31.45
Kerala 19 19 Jan 1.21 217.40 337.34
Maharastra 97 24 Feb 3.71 250.76 4.53
Madhya Pradesh 33 10 March 2.81 266.60 335.78
Nagaland 1 1 April 1.86 48.94 357.97
Orissa 13 10
Pondicherry 3 3
Punjab 12 6 damage distance for heat flux of 37.5 kW/m2 will have about
Rajashthan 54 17 283 m radius and for heat flux of 4 kW/m2, it will be about
Tamilnadu 41 31 832 m. The total time duration would be about 1 hour. Hence,
Uttar Pradesh 40 14
West Bengal 40 23 serious damage will occur to process equipments within
283m radius zone. This will also have a disastrous effect on
It is observed from this analysis that there may be 100% the work-force in that area.
fatality within a radius of 350 m from this coke-oven stor- It is observed from this analysis that there may be 100%
age tank. In the remaining band damage due to fire burn in- fatality within a radius of 283 m from blast furnace storage
juries of varying degrees may result from thermal radiation. tank. In the remaining band, damage due to fire burn inju-
A jet fire is usually a localized, but very destructive to ries of varying degrees may result from thermal radiation.
anything close to it. This is partly because of producing ther- The jet fire/flare may have serious domino and cascad-
mal radiation; the jet fire causes considerable convective ing effects on the storage/process areas of flammable chemi-
heating in the region beyond the tip of the flame. The high cals, triggering off fire and explosion, the effect of which
velocity of the escaping gas entrains air into the gas ‘jet’ may travel off-site. Thus, preparedness is required to effec-
causing more efficient combustion to occur than pool fires. tively handle the off-site emergency for adequate fire-fight-
Although damage distances are calculated, based on ther- ing, rescue, evacuation and medical treatment for burn inju-
mal radiation level, the degree of injury also depends upon ries.
the duration of exposure. But, it is worth mentioning in this .

context that human beings are also vulnerable to prolonged

exposure to low-level heat flux or thermal radiation.
The jet fire/flare may have serious domino and cascad-
ing effects on the storage/process areas of flammable chemi-
cals, triggering off fire and explosion, the effect of which
may travel off-site.
Jet fire/flare from major leakage of blast furnace gas in
blast furnace storage tank: About 1,00,000 cu.m blast fur-
nace gas is stored in one gas holder at an approximate pres-
sure of 350 mm water gauge.
The assumptions of this scenario are heavy leakage of
gas due to rupture of out-going/ incoming pipe due to sabo-
tage, riot, corrosion, etc., and getting ignited from an
external source.
It has been estimated by applying CHARM software that
It has been observed from this study that there may be 100%
fatality within a radius of 350 m for the coke-oven storage
tank, and 283 m for blast furnace storage tank. In the re-
maining band for both the coke-oven and blast furnace stor-
age tank, damage due to fire burn injuries of varying de-
grees may result from thermal radiation. Followed by emer-
gency plan preparations, regular mock-drills may provide
enough preparedness required to avert a major crisis. Com-
pliance to regular safety audits is necessary in addition to
other mandatory requirements. Maintenance of proper work-
ing condition is necessary. Consequence analysis could help
manufacture plants, local governments and communities to
address needs for improving their mitigation facilities and
warning systems for disaster prevention and protection pro-
grams. Moreover, government should regulate and mandate

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coke-oven gas storage tanks and blast furnace storage tank
to execute their consequence analysis in order to identify
problems in advance and to lessen possible impacts on pub-
lic safety. A good communication system, regular interac-
tion between various departments, education and safety train-
ing of the employees, review of past accidents/reports/pub-
lished safety reviews, daily site inspection and ready avail-
ability of emergency equipment are the key areas of effec-
tive off-site emergency preparedness. Since, the disaster may
result in burn injuries, upgradation of medical facilities and
the associated infrastructure may be necessary to mitigate
the effect of the crisis, particularly for the area under study.

. 0 - 
The author is grateful to the Director, Central Mechanical
Engineering Research Institute, Durgapur for his constant
encouragement and active support. Author is also grateful
to Dr P. K. Chatterjee, Scientist & Head, Thermal Engg.
Group, CMERI, Durgapur for his inspiration. Author is also
thankful to the Department of Environment, Govt. of West
Bengal, Kolkata for their financial support to carry out this
study and West Bengal Pollution Control Board, Regional
Office, Durgapur for providing the meteorological data.
 1   
AICE 1989. Guidelines for Chemicals Process Quantitative Risk Analysis.
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Biswajit Ruj
American Institute of Chemical Engineers.
Ditali, S., Colombi, M., Moreschini, G. and Senni, S. 2000. Consequence
analysis in LPG installation using an integrated computer package. J.
Hazard. Mater., 71(1-3): 159-177.
ILO 1990. Major Hazard Control-A Practical Manual. An ILO Contribu-
tion to the International Programme on Chemical Safety on UNEP,
ILO, WHO (IPCS), International Labour Organisation, 2nd ed.
Khan, F.I. and Abbasi, S.A. 1999. Assessment of risks posed by chemical
industries - Application of a new computer automated tool MAXCRED-
III. J. Loss Prev. Proc. Ind., 12(6): 455-469.
Raghavan, K.V. and Swaminathan, G. 1996. Hazard Assessment and Dis-
aster Mitigation in Petrochemical Industries. Oxford Publishing Com-
pany, Chennai.
Ruj, B., Rehman, I. and Bandyopadhya, A.K. 2006. Off-site emergency
scenario, a case study from a LPG bottling plant. J. Loss Prev. Proc.
Ind., 19: 645-647.
Ruj, B., Chatterjee, P.K., Rehman, I. and Ray, B.C. 2009. Consequence of
hazards on some petroleum storage tanks and model for off-site emer-
gency plan. J. Disaster Advances, 2(2): 36-40.
Ruj, B. 2005. Emergency planning-A tool for the mitigation of industrial
explosion. Nat. Environ. Pollut. Tech. 4(1): 123-126.
Ruj, B. and Chatterjee, P.K. 2007. Industrial disaster management and emer-
gency plan. International Seminar on Dangerous Trade: Histories of
Industrial Hazards Across a Globalizing World, Stony Brook Univer-
sity, New York, United States of America.
Spadoni, G. and Uguccioni, G. 2003. The new version of ARIPAR and the
benefits given in assessing and managing major risks in industrialized
areas. Trans Ichem E, Pt B, Proc. Safety Environ. Protec., 81(1):
19-30.
Tixier, J., Dussere, G., Doumax, S.R., Ollivier, J. and Bourely, C. 2002.
OSIRIS: Software for the consequence evaluation of transportation of
dangerous goods accidents. Environ. Model Software, 17(7): 627-637.
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*Malviya National Institute of Technology, Jaipur-302 017, Rajasthan, India

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,1752'8&7,21 apparent, more attention is being placed on how treatment

As freshwater becomes increasingly scarce, it is necessary
for policy makers and leaders to shift attention to alternative
sources of water. Faced with the twin problems of water scar-
city and limited access to safe alternative sources of water,
some countries and many of their citizens are turning to-
wards a formal recognition of the role that wastewater use
can play in supplementing existing sources of water
(Hernandez Leal et al. 2010).
People are now waking up to the benefits of grey water
reuse, and the term “wastewater” is in many respects a mis-
nomer. May be a more appropriate term for this water would
be “used water”. Grey water refers to the wastewater that
comes from kitchens, bathrooms and laundry. While bath-
room and laundry waters are relatively benign, kitchen wa-
ter deserves special attention since it is loaded with organic
matter from food waste. Grey water is distinct from black
water (that comes from the toilet) as there are fewer health
and environmental risks associated with its use (Devine et
al. 1998). It is estimated that 55%-65% of household water
effluent is grey water (Burnat 2007, Diener & Morel 2006).
Grey water including its separation, containment and use, is
a simple, home-based Water Demand Management (WDM)
strategy that has benefits at the household level as an alter-
native water resource to optimize productivity, if used wisely
and appropriately. Grey water after proper treatment can be
used for various purposes like irrigation, urinal flushing, etc.
Reuse of grey water serves two purposes: reduces fresh wa-
ter requirement and reduces sewage generation (U.S. EPA
2004). As awareness of the potential and challenges associ-
ated with grey water recovery and use have become
and use at the household level can be promoted. Grey water
fundamentally preserves the existing freshwater supply, and
in that way is a significant WDM strategy. Estimates of the
proportion of household wastewater, that is grey water, usu-
ally vary from 65% up to 80% (Burnat 2007). No matter the
amount, its use conserves water supply by negating the need
to acquire water from the municipal network, or, in unserved
areas, from private vendors.
Grey water treatment options: Grey water reuse methods
can range from low cost methods such as the manual buck-
eting of grey water from the outlet of bathroom to primary
treatment methods that coarsely screen oils, greases and sol-
ids from the grey water before irrigation via small trench
systems, to more expensive secondary treatment systems that
treat and disinfect the grey water to a high standard before
using for irrigation. The choice of system depends on a
number of factors including whether a new system is being
installed or a disused wastewater system is being converted
because the household has been connected to sewer.
Slow sand filters are probably the most effective, sim-
plest and least expensive water treatment process for devel-
oping countries (www.epa.gov) . They require few techni-
cal components and usually no chemicals. They consist of
fine sand supported by gravel. They capture particles near
the surface of the bed and are usually cleaned by scraping
away the top layer of sand that contains the particles
(CPHEEO 1991).
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To evaluate the potential of slow sand filter as treatment

option for grey water, sand filter model was fabricated at
PhE Lab, MNIT, Jaipur. The specifications for the designed
sand filter are:
Filter unit: The filter unit was fabricated in the Hydraulics
Laboratory in MNIT, Jaipur. It was fabricated using 2mm
galvanized iron sheets. The sheet was welded to form a hol-
low cylinder with an inside diameter of 43 cm. Mseal adhe-
sive was used to avoid leakages. Its dimensions were: 0.50
m height and 0.43 m diameter and it was checked that there
were no leaks by filling it with water.
Filter media: Sand which served as filter media was prop-
erly washed and cleaned before it was put in the filter. The
sand was obtained locally and sieved to obtain a media with
required sand size. Sand of effective size 0.2 mm and uni-
form coefficient 2.5 was used.
Base material: Gravels were used as base layer. The layer
of sand was supported on gravel, which permits the filtered
water to move freely to the under drains. The supporting
gravel was washed out and placed in three layers. Table 1
describes the specifications of the base material used. The
total layer was divided into three parts with the topmost layer
of 23 mm depth and size of gravel being 2 to 4.75 mm,
whereas the bottom layer of depth 23 mm and size of gravel
being 40 to 63 mm.
Under drainage system: The filter media and the base ma-
terials were supported over the under drainage system which
eventually collects the filtered water and delivers it to the
clean water reservoir. Perforated pipe was used for under
drainage system.
352&(66 '(6&5,37,21
The Fig. 1 describes the line diagram of the process involved.
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I = Topmost layer of gravels
II = Intermediate layer of gravels
III = Bottom layer of gravels
F1 = Feeding reservoir 1
Feeding reservoir 2
Fig. 1: Diagram of fabricated model for grey water treatment.
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Nidhi Poonia and M. K. Jat
In first feeding reservoir (F1) the sample was put. From this
F1 tank, an outlet pipe was connected to second tank (F2)
through floating ball valve. This floating ball was used to
maintain the head in F2 basin. From F2 basin the sample
was sent to filter unit (drum). On the top of the filter unit
perforated plate was fixed so that the sand layer was not dis-
turbed with the direct flow of water. Through this, it was
also assured that the schmutzdecke was not disturbed by the
turbulence of water. For the under drainage system, perfo-
rated pipe was used. This perforated pipe was connected to
the outlet pipe for sample collection. Samples of both the
feed and product water were collected for analysis to assess
treatment performance.
First, with all the outlet valves closed, the filter was
charged with clean water, introduced from the bottom to a
level of about 10 cm above the sand bed. This was done to
drive out the air bubbles from the filter bed and then the
inflow was started.
The influent and the effluent samples from the sand fil-
ter were analyzed for different physical and chemical pa-
rameters like alkalinity, chloride, hardness, nitrate, turbid-
ity, TSS, TDS, pH, BOD and COD using standard methods
(APHA, AWWA, WPCF 1981).
5(68/76$1'',6&866,21
Table 2 shows the consolidated values of influent and efflu-
ent of grey water treatment.
The average value of alkalinity in the influent ranged
from 238 mg/L to 865 mg/L with laundry water having
higher values due to the use of detergents, whereas the alka-
linity levels in the effluent ranged from 178 mg/L to 416
mg/L. Composite water and bath water showed the little re-

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 CHARACTERIZATION AND TREATMENT OF GREY WATER FOR RECYCLING
duction in the values with reduction of more than 20%;
kitchen water and wash basin water showed the moderate
reduction between 20-50%, and the laundry water showed
the maximum reduction of more than 50%. According to IS:
3307, in the wastewater to be discharged onto land for irri-
gation, the chloride value should be 600 mg/L, whereas ac-
cording to IS: 2490 standards for wastewater to be discharged
into inland surface water it is 1000 mg/L. The values of chlo-
ride in the influent have ranged from 29 mg/L to 62 mg/L,
whereas the effluent levels have ranged from 23 to 49 mg/L.
The average reduction of hardness in the kitchen, laundry
and bath water samples showed the highest reduction per-
centage of more than 50%, whereas the wash basin water
showed the moderate reduction between 20-50%. Turbid-
ity, one of the most important parameters to monitor the per-
formance of filter, has ranged from 24 ppm to 77 ppm in the
feed water, whereas the effluent levels have ranged from
4ppm to 33ppm. The average values of nitrate in the feed
water have ranged from 32.54 mg/L in the wash basin water
to 66.69 mg/L in laundry water, whereas the average efflu-
ent values have ranged from 22.61 mg/L in kitchen water to
45.26 mg/L in laundry water.
According to IS: 3307, the BOD in wastewater to be dis-
charged onto land for irrigation is 500mg/L. The average
value of the BOD in the influent ranged from 196.07 mg/L
in the composite sample to 310.71 mg/L in laundry water.
The average values of COD in the effluent ranged from
183.24 mg/L in kitchen water to 350.77 mg/L in wash basin
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water. The average percent reduction of TSS was
moderate with values ranging from 20-50%. The average
values of pH in the effluent ranged from 7.4 in laundry wa-
ter to 7.7 in bath water.
Fig. 2 shows the average influent and effluent concen-
tration of different physical and chemical parameters in
kitchen water. Turbidity showed the highest reduction of
61.28 % followed by hardness with 51.25 %. The pH reduc-
tion was of not significant. Laundry water average influent
and effluent concentrations of different parameters are shown
in Fig. 3. Total dissolved solids showed the highest peak
due to the presence of detergent products dissolved in sam-
ple. The highest reduction was of turbidity with 69.87%.
High alkalinity values were noticed due to use of detergents
and soap products.
Figs. 4 and 5 show the average concentrations of differ-
ent parameters in bath water and wash basin water respec-
tively. The percent reduction of TSS in bath water was 42%,
whereas in wash basin water it was 31%. There was 73%
reduction of turbidity in wash basin water, whereas it was
64% in bath water. Approximately 50% reduction of hard-
Table 1: Base material specifications (depth and size).
Layers Depth (mm) Size (mm)
Topmost layer (I) 23 2 to 4.75
Intermediate layer (II) 23 4.75 to 40
Bottom layer (III) 23 40 to 63
 Nidhi Poonia and M. K. Jat

Table 2: Average influent and effluent values. St Dev = Standard Deviation

Parameter Kitchen water Laundry Water Bath water Wash basin water Composite sample
Average Average Average Average Average Average Average Average Average Average
feed product feed product feed product feed product feed product
(± St Dev) (± St Dev) (± St Dev) (± St Dev) (± St Dev) (± St Dev) (± St Dev) (± St Dev) (± St Dev) (± St Dev)

Tot. alkalinity 238.57 180.95 865.7 416.71 264.2 228.27 258.5 178.57 305.78 263.94
(±10.3) (±11.01) (±11.53) (±7.92) (±9.6) (±6.29) (±4.96) (±8.03) (±8.99) (±4.1)
Chloride 29.01 25.09 31.94 23.61 62.55 49.7 34.19 24.81 45.72 38.47
(±1.5) (±2.3) (±4.16) (±2.07) (±5.24) (±3.9) (±2.81) (±1.68) (±4.03) (±4.07)
Hardness 200.42 97.69 350.28 167.91 180 89.37 188.2 110.23 181.42 166.07
(±10.02) (±3.45) (±7.2) (±5.64) (±4.37) (±5.70) (±5.68) (±5.68) (±5.06) (±3.34)
Turbidity 24.42 4.57 54.15 16.42 45.42 16.28 34.85 9.28 77.71 33.14
(±2.77) (±2.38) (±2.58) (±2.12) (±2.55) (±3.69) (±3.6) (±1.82) (±2.81) (±2.41)
Nitrate 34.52 22.61 66.69 45.26 59.51 38.74 32.54 28.35 57.99 47.96
(±3.56) (±2.29) (±3.62) (±2.43) (±1.20) (±4.64) (±2.44) (±2.03) (±1.82) (±2.45)
BOD 224.42 168.45 310.71 220.41 222.85 183 302.14 214.35 196.07 158.27
(±4.03) (±4.90) (±7.75) (±3.93) (±3.39) (±3.98) (±7.95) (±5.59) (±6.52) (±5.19)
COD 265.26 184.24 565.42 303.27 298.97 198.63 520.85 350.77 399.513 33.31
(±3.24) (±7.88) (±7.15) (±4.95) (±5.89) (±4.69) (±5.18) (±4.02) (±6.41) (±4.63)
TSS 1.37 1.18 3.15 1.82 1.91 1.09 2.95 2.03 1.16 0.31
(±0.37) (±0.41) (±0.53) (±0.95) (±0.79) (±0.80) (±0.13) (±0.46) (±0.18) (±0.17)
TDS 451.42 357 4237.8 3108 355.14 242.85 546.85 383.71 786.28 539.28
(±6.92) (±4.86) (±3.68) (±5.31) (±4.70) (±5.19) (±5.54) (±4.16) (±3.7) (±5.14)
pH 7.7 7.5 7.9 7.4 8.05 7.7 7.5 7.2 7.9 7.5
(±0.13) (±0.05) (±0.18) (±0.22) (±0.10) (±0.17) (±0.07) (±0.10) (±0.18) (±0.09)

The units are in mg/L except pH and turbidity.

ness was noticed in both the samples. Percentage reduction
of pH in both the samples was of no significance. There was
low reduction of BOD in both the samples with 17% in bath
water and 29% in wash basin water.
&21&/86,216
The model was fabricated successfully and run for steady
state condition. Samples of kitchen water, laundry water, bath
water and wash basin water were collected from different
sites from MNIT, Jaipur campus and analysed for physical
and chemical parameters. There was high variation in the
quality of grey water due to factors such as water use
efficiencies of appliances, individual habits, products used
(soaps, shampoos, detergents) and other site specific char-
acteristics. It was found that the common contaminants in
the kitchen water are suspended solids, which are added due
to the presence of food particles in the water. They have to
be screened out before treatment to avoid any blockages. Bath
water and basin water was mainly contaminated by suspended
particles like dirt, lint and hair. There was a sharp increase
in the pH value. During the preliminary runs the filtrate ob-
tained showed the negligible percentage removal in contami-
nants. This was due to the reason that the filter was filled
with the freshly cleaned sand and thus there was no biologi-
cal layer developed on the sand bed. It was found that there
is a small decrease in the pH in the filtrate. This is probably
due to dissolution of carbon dioxide as a result of biological
activity in the top layer.
5()(5(1&(6
APHA, AWWA, WPCF, 1981. Standard Methods for the Examination of
Water and Wastewater, 15th ed., American Public Health Association,
Washington DC.
Burnat, Jamal 2007. Evaluation of On-Site Gray Wastewater Treatment
Plants Performance in Bilien and Biet-Diko Villages/Palestine. Con-
ference Paper for the Greywater Stock-Taking Meeting, IDRC-CSBE.
Devine, B. 1998. Greywater recycling in Western Australia. Water Journal
of the Australian Water and Wastewater Association, Artarmon, NSW,
25(2).
Hernandez Leal, L., Temmink, H., Zeeman, G. and Buisman, C.J.N. 2010.
Comparison of three systems for biological greywater
treatment. Water, 2: 155-169.
Li, F., Gulyas, H., Wichmann, K. and Otterpohl, R. 2009. Treatment of
household grey water with a UF membrane filtration system.
Desalination and Water Treatment, 5: 275-282.
Morel, Antoine and Stefan Diener 2006. Greywater Management in Low
and Middle-Income Countries. Water and Sanitation in Developing
Countries (Sandec). Eawag, Swiss Federal Institute of Aquatic Sci-
ence and Technology.
Onsite/Decentralized Treatment Technologies Fact Sheet. http://
www.epa.gov/owm/decent/tech_right.htm
U.S. EPA 2004. Manual of Guidelines for Water Reuse, EPA/625/R- 9/
2004, U.S. Environment Protection Agency and U.S. Agency for In-
ternational development, Washington DC.
Water Supply and Treatment Manual 1991. Central Public Health and En-
vironmental Engineering Organization.

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Ratan Chand Sharma and Shivani Dogra
Department of Chemistry, Post Graduate Government College, Hamirpur-177 001, H. P., India

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,1752'8&7,21
Soil generally refers to the loose material composed of
weathered rock and other minerals, and the partly decayed
organic matter that covers large parts of earth (Wild 1996).
Soil is a reservoir of nutrients in one form or other and dif-
fers from the parent materials in the morphological, physi-
cal and biological properties. The quantity of potassium is
plentiful except in sandy soils. Calcium shows great varia-
tion, but it is generally present in lesser amount than potash.
Phosphorus is needed not only as nutrients but also as a means
of controlling soil acidity. Soil respiration is augmented by
the respiration of living plants roots and is further abun-
dance of carbonaceous materials in the soil (White 1987).
Soil needs specific heat of soil, heat vaporization, and thus
it is transferred through the process of conduction (Jaiswal
& Wadhwani 1980). Further, calcium, magnesium, potas-
sium and sodium reduce the acidity and increase alkalinity
(Brady 1984).
Soil plays a pivotal role in supporting the growth of crops,
and other vegetation maintaining the environment clean, and
acts as source and sink for atmospheric gases. Man depends
on soils and to a certain extent good soils are dependent upon
the man and the use he makes of them. Soils are natural bod-
ies on which plant grow.
Soils in India can be classified into the following three
groups on the basis of cultivation (Jaiswal & Wadhwari
1980).
• Alluvial, black and red soils, which are fertile, arable and
cultivable.
• Peat and Marsh, which are saline, alkaline and poten-
tially arable.
• Laterite and forest and hill soils, which are not suitable
for cultivation.
Alluvial type of soil is deposited by the action of mov-
ing water (usually rivers) and is usually highly fertile. This
soil has texture with favourable physical characteristics and
good permeability. Small showers are useful and there is
utilization of the water held by soil due to low moisture
content at wilting point. These include the deltaic alluvium,
calcareous alluvial soil, coastal alluvium and sands. It is the
largest share to the agriculture wealth. The soil contains vary-
ing amounts of calcium carbonate (CaCO3) and soluble salts,
and is neutral to alkaline. The lime content increases at lower
depths. They are generally poor in phosphorus pentaoxide
(P2O5), nitrogen and organic matter.
Black soil is highly argillaceous (clayey), fine grained
and dark, and contains a high proportion of calcium and mag-
nesium carbonates. The black soil is deeper, clay to clayey
loam and characterized by low permeability and high water
holding capacity, low infiltration rate, high plasticity and
stickness, low organic matter, high CEC, calcareous nature
and slightly old aline. It is specially suited for cotton culti-
vation. In the valleys of the rivers Tapti, Narmada, Godavari
and Krishna, heavy black soil is often six metres deep.
Red soils are light textured, shallow to medium in depth
and usually under layer by compact subsoil, fairly porous
with low water holding capacity. They are generally poor in
nitrogen, phosphorus and humus, and are present in many
areas of Tamil Nadu, Karnataka, Goa, Daman and Diu, south-
eastern Maharashtra, Madhya Pradesh, Orissa and
Chhotanagpur plateau.
Laterite soil is formed by the decomposition of many
 Ratan Chand Sharama and Shivani Dogra

kinds of rocks, and found especially in tropical rain forests.
It is a vesicular rock composed essentially of mixture of hy-
drated oxides of aluminium and iron with small amount of
manganese dioxide, titanium oxide, etc. It is common in the
low hills of Andhra Pradesh, Karnataka, Kerala, Madhya
Pradesh and Orissa. The laterite may be broken off and car-
ried to lower level by streams and when deposited may get
into a compact may be the segregative action of the hydrated
oxides.
Area under investigation: Himachal Pradesh lies in the
western Himalayas between 30°22’40” to 33°12’40” north
latitude and 75°47’55” to 79°04’20” east longitude (Fig. 1).
Its geographical area is 55,673 sq. km, which is 1.69% of
the India’s total area. The snowy mountains of Himachal are
known for its natural and scenic beauty of Dhauladhar, Pir
Panjal, Shivalik, great Himalayan and Zaskar ranges. It is
one of the most complicated geological regions of Himala-
yas. The region falls in four broad stratigraphical zones: 1.
Shivaliks; 2. Inner Himalayas; 3. Great Himalayas; 4. Trans
Himalayas.
Inner Himalayas lie between the main boundary thrust
and central Himalayan thrust. Main boundary thrust lies
between middle Himalayas and lower Himalayas. The cen-
tral Himalayan thrust lies between the greater Himalayas and
middle Himalayas. The granite and crystalline rocks pre-
dominate in the zones, which are of unfossiliferous rudi-
ments. The Karol belt stretches from the Shimla region to-
wards the earth and repartees this region from the Shivalik

Fig. 1: Location map of the area.

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 CHARCTERIZATION OF THE SOIL OF LOWER HIMALAYAS OF HIMACHAL PRADESH
system. The paleozoic region in the lower Himalayas is sud-
denly interrupted by the transgression of the permo-
carboniferous with an outstanding different Gondwana rock
sequence.
Outer or sub-Himalayas are also known as Manak Parbhat
in the ancient time. The Shivalik hill tack runs along the
southern border of the State from Kangra to Sirmour. There
are the outermost ranges separating it from the Punjab plain.
The altitude ranges from 600 metres to 120 metres. These
ranges are youngest of Himalayan ranges and made up of
tertiary sediments consisting of sand and clay. This range in
Himachal is not a continuous range but forms several minor
ranges. They are seen as eroded hill masses or residual de-
tached hillocks. The crest line on some important Shivalik
ranges are known by several local names like Dhog Dhar in
Sirmaur, Ramgarh Dhar in Una, Chaumukhi range, Dharti
Dhar, Sikandar Dhar and Naina Devi Dhar in Hamirpur and
Bilaspur districts and Panchmunta hill in Solan district. The
middle or lesser Himalayas are located to the north of
Shivalik range. They form an intricate and rugged mountain
system, about 60-80 km wide and 1000-4000 metres high,
with several peak sites to nearly 5000 metres, and remains
snow covered throughout year.
Inner Himalayas are not a continuous range but consist
of a number of ranges like Dhauladhar, Pir Panjal, Churdhar
and Shimla ranges. Dhauldhar or ‘White Mountains’ is the
first prominent and outermost Inner Himalayas range that
overlooks the Kangra valley. Dhauladhar runs parallel to the
Pir Panjhal running across Kangra District. The three major
rivers, Satluj near Rampur, Beas at Larji and Ravi near
Chamba cut the Dhauladhar range forming deep gorges. The
range is most prominent in Kangra valley where it records a
3600 metres vertical rise within a short distance of only 11
km. Bottom of the valley is situated at about 900 metres above
sea level.
Pir-Panjal or the Pangi range is next range of inner Hima-
layas range. It enters Himachal Pradesh from Kashmir run-
ning through Chamba or a water divide between Chenab in
the north and Ravi and Beas rivers in the south. The range is
known for its great height and complex topography and has
number of glaciers.
Nag-Tibba range is the outermost of the inner Himalayas
range in eastern part of Himachal. Shivalik Duns
intermittently located between the Shivalik and lesser
Himalayas are the duns, which are flat-bottomed longitudinal
structural valleys within own drainage systems. The Shivalik
hills gently slope into flat bottomed strike. Valleys locally
called ‘duns’ towards the north. Some important dun valleys
of Himachal Pradesh are Paunta valley of Sirmaur, Kangra
valley, Nalagarh dun in Solan and Jaswan dun in Kangra
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

district. Soil samples from Kangra, Hamirpur, Mandi and
Bilaspur districts were analysed.
Heavy metal contamination of soils is a worldwide prob-
lem of increasing importance and great environmental con-
cern (Alloway 1990, Kabata Pendias & Pendias 1992). Soil
contamination by heavy metals results in some changes in
the counts of microorganisms and enzymatic activity which,
according to Dick et al. (2000), Kucharski (1997) and Treasar
Ceped et al. (2000) is an objective measure of the soil’s
microbiological status. The negative influence of most of
the heavy metals on the activity of soil enzymes was re-
ported by Leiros et al. (1990), Nowak & Smolik (2002),
Nowak et al. (2003) and Wyszkowska & Kucharski (2003).
Three metals often cited for their deleterious effects on both
human and plant metabolism are cadmium (Cd), lead (Pb)
and zinc (Zn). The excessive uptake of these elements by
crop plants from the soil creates problem of crop yield re-
duction due to inhibition of metabolic processes in plants
(Foy et al. 1978, Costa & Morel 1993, Aery & Jagetiya 1997).
In addition, the crops so contaminated serve as a source of
heavy metals in our food supply (Somers 1974, Singh et al.
1995, Sajwan et al. 1996). Therefore, the problems posed by
increased amounts of these metals in the environment de-
mand a thorough understanding of their phytotoxic effects
in a soil plant system.
0$7(5,$/6$1'0(7+2'6
The pH was measured by a pH meter and organic matter by
Walkley and Black wet digestion method. The potassium
determination was carried out by flame photometer. The
micronutrients (zinc, copper, iron, manganese) were deter-
mined by diethylene triamine penta-acetic acid (DTPA) ex-
traction method. The dissolved elements in the extract were
then measured by Atomic Absorption Spectrophotometer.
5(68/76$1'',6&866,21
The negative effect of heavy metals on the soil, which was
observed in the present work has confirmed and verified in
literature (Leiros et al. 1990). Furthermore, Welp (1999)
claims that copper along with chromium III and VI belong
to metal which has adverse effects on the activity of enzymes
in soil. Leiros et al. (1990) found that copper needed to be
applied at concentration as high as 800 mg/kg to produce an
unfavourable effect on the activity of enzymes. As a conse-
quence of these industrial activities in Barmana, large
amounts of wastes have been generated. These wastes con-
tain heavy metals such as copper, zinc, cadmium, lead, mer-
cury and arsenic at level that exceeds the critical European
ones (Garica et al. 2001b).
The physico-chemical characteristics of soil such as pH,
 Ratan Chand Sharama and Shivani Dogra

Table 1: pH values for different soil samples. Table 3: Values of available potassium for different soil samples.

Sample pH Sample No. kg/h Rating

Kangra 6.8 Kangra 649.6 H

Lambloo 7.8 Lambloo 672.4 H
Sandhol 7.0 Sandhol 358.4 H
Ghagas 8.0 Ghagas 268.8 H
Dharamshala 7.1 Dharamshala 1366.4 M
Bhota 8.0 Bhota 89.6 L
Ratti 7.2 Ratti 89.5 L
Kularu 7.8 Kularu 134.4 M
Yol 6.6 Yol 268.8 M
Nohangi 7.4 Nohangi 918.4 H
Dandor 7.4 Dandor 201.6 M
Dangar 8.3 Dangar 67.2 L
Chamunda 7.2 Chamunda 291.2 H
Sujanpur Tira 7.0 Sujanpur Tira 44.8 L
Sundernagar 8.1 Sundernagar 33.6 H
Barmana 8.1 Barmana 918.4 H

Table 2: Values of organic carbon for different soil samples. Table 4: Values of micronutrients for different soil samples.

Sample X-Y (mg) Percentage(%) Rating Sample No. Zn Cu Fe Mn

of Organic Carbon (ppm) (ppm) (ppm) (ppm)

Kangra 6.8 0.0 L Kangra 3.48 2.13 4.05 1.72

Lambloo 7.9 1.1 H Lambloo 2.33 1.20 7.11 0.78
Sandhol 3.8 1.0 M Sandhol 2.47 0.66 4.49 0.9
Ghagas 2.5 0.6 M Ghagas 0.6 0.78 8.08 9.56
Dharamshala 4.6 0.7 M Dharamshala 3.98 0.53 26.33 2.58
Bhota 4.2 0.6 M Bhota 0.80 0.3 5.8 0.92
Ratti 2.2 0.5 M Ratti 0.68 0.8 4.93 0.9
Kularu 5.2 1.3 H Kularu 1.3 1.0 4.0 13
Yol 9.0 1.3 H Yol 0.69 1.63 3.5 0.4
Nohangi 10.8 1.6 H Nohangi 0.62 1.0 5.72 0.4
Dandor 8.8 2.2 H Dandor 2.2 0.2 3.82 11.73
Dangar 5.2 1.3 H Dangar 1.3 0.1 6.25 1.88
Chamunda 5.4 0.8 M Chamunda 3.77 1.25 21.42 1.21
Sujanpur Tira 5.6 1.4 H Sujanpur Tira 0.7 0.7 4.2 0.85
Sundernagar 16.6 4.3 H Sundernagar 4.3 3.22 7.67 0.86
Barmana 2.4 0.6 M Barmana 0.4 0.70 7.6 23.0

organic matter, potassium and micronutrients like zinc, cop-
per, iron and manganese are given in Tables 1, 2, 3 and 4,
and Figs 2-29.
The heavy soil contamination by copper, manganese and
organic matter was reported in Sundernagar Barmana belt
that is 3.22 ppm, 23 ppm and 4.3% respectively, which is
responsible for pollution of the soil in the area. These metals
have negative effect on the activity of soil enzymes like de-
hydrogenase, urease acid phosphatase and alkaline phos-
phatase, etc. There is, thus, inhabitation of soil enzyme ac-
tivity as reported by Nowaki (2002). Furthermore, Welp
(1999) claims that copper along with chromium VI and III
belongs to metals which has strong adverse effect on activ-
ity of enzyme. The enzymes are contaminated by copper and
other heavy metals which in turn affect different sorptive
properties of soil, as reported by the Leiros et al. (1990) that
excessive copper can affect the activity of enzymes in soil.
Therefore, it was suggested that the copper is lesser toxic
than the zinc. But, since both of them are reported equally in
all the soil samples, it is necessary to avoid the concentra-
tion of copper as well as zinc. The iron, manganese and po-
tassium have concentrations of 26.33 ppm, 23.0 ppm and
1366.4 kg/ha was reported in Dharamshala and Barmana.
This high concentration may be attributed to the solid waste
and mining activities in the area. Thus, therefore, it becomes
pertinent that the soil may be saved from such activities. The
highest pH was reported at Dangar towards more basic due
to excessive limestone mining in the area for the use of ce-
ment and other industries. Therefore, it is suggested that if
this excessive mining is not checked, the day is not far off

9RO1Rx
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 CHARCTERIZATION OF THE SOIL OF LOWER HIMALAYAS OF HIMACHAL PRADESH 

Fig. 2: Variation in the value of Fig. 3: Variation in the value of Fig.4: Variation in the value of Fig. 5: Variation in the value of pH
pH at different sites in Kangra pH at different sites in Hamirpur pH at different sites in Mandi at different sites in Bilaspur
district. district. district. district.

Fig. 6: Variation in the value of Fig. 7: Variation in the value of Fig. 8: Variation in the value of Fig. 9: Variation in the value of
organic matter at different sites in organic matter at different sites in organic matter at different sites organic matter at different sites in
Kangra district. Hamirpur district. in Mandi district. Bilaspur district.

Fig. 10: Variation in the value of Fig. 11: Variation in the value of Fig. 12: Variation in the value of Fig. 13: Variation in the value of
potassium at different sites in potassium at different sites in potassium at different sites in potassium at different sites in
Kangra district. Hamirpur district. Mandi district. Bilaspur district.

Fig.14: Variation in values of Fig.15: Variation in values of Fig.16: Variation in values of Fig.17: Variation in values of
micronutrient zinc at different micronutrient zinc at different micronutrient zinc at different micronutrient zinc at different
sites in Kangra district. sites in Hamirpur district. sites in Mandi district. sites in Bilaspur district.

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Ratan Chand Sharama and Shivani Dogra

Fig. 18: Variation in values of Fig. 19: Variation in values of Fig. 20: Varition in values of Fig. 21: Variation in values of
micronutrient copper at different micronutrient copper at different sites micronutrient copper at different micronutrient copper at different
sites in Kangra district. in Hamirpur district. sites in Mandi district. sites in Bilaspur district.

Fig. 22: Variation in values of Fig. 23: Variation in values of Fig. 24: Variation in values of Fig. 25: Variation in values of
micronutrient iron at different micronutrient iron at different sites in micronutrient iron at different micronutrient iron at different sites
sites in Kangra district. Hamirpur district. sites in Mandi district. in Bilaspur district.

Fig. 26: Variation in values of Fig. 27: Variation in values of Fig. 28: Variation in values of Fig. 29: Variation in values of
micronutrient manganese at micronutrient manganese at different micronutrient manganese at micronutrient manganese at
different sites in Kangra district. sites in Hamirpur district. different sites in Mandi district. different sites in Bilaspur district.

when soil will have excessive basic nature which can greatly
affect fertility of soil and water quality.
Both, water and soil play a significant role in ecosys-
tems and plant growth but, when their quality is not good,
they inhibit plant growth. Water is necessary for all living
beings for their sustenance. The polluted water disturbs the
growth of animals and plants.
Organic carbon plays an important role in soil biology
by releasing nutrients to plants, providing buffering to soil,
and stabilising the soil structure. In fact, these properties
along with organic carbon, nitrogen and phosphorus are con-
sidered critical indicator for the health and quality of soil.
The organic carbon was found to be maximum in
Sundernagar (4.3%) district of Mandi, and minimum in
Kangra (0.0%). The high content of 4.3 % was attributed to
the smoke released by cement plant located in the area. Loss
of organic carbon lead to the reduction in soil fertility, land
degradation and even desertification due to the CO2, SO2,
CO and unburnt carbon. The unburnt carbon generally in-
creases the pH of soil and, thus, affect the fertility. Also the
ash with unburnt carbon decreases the absorption capacity

9RO1Rx
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 CHARCTERIZATION OF THE SOIL OF LOWER HIMALAYAS OF HIMACHAL PRADESH
of the soil. Excessive amount of organic carbon can be prob-
lematic.
Potassium is one of the important nutrients found in soil.
Potassium is high in most of the soils, except in those con-
taining sand. Potassium is important in plant photosynthetic
process and in helping plants to metabolize their food to get
more energy. Potassium control water and chemicals inside
plants and helps plants to function well. The potassium in
soil varied from 67.2 kg/ha (Dangar of District Bilaspur) to
1366.4 kg/ha (Dharamshala of District Kangra). The per-
missible limit of potassium is 118 kg/ha to 280 kg/ha.
Therefore, it was established that some places have potas-
sium content lower than the permissible limit, i.e., Sujanpur
Tira (44.8 kg/ha), Dangar (67.2 kg/ha), Bhota (89.6 kg/ha)
and higher than the permissible limit like Chamunda (291.2
kg/ha), Sundernagar (336 kg/ha), Sandhol (358.4 kg/ha),
Lambloo (672.4 kg/ha), Kangra (49.6 kg/ha), Nohangi (918.4
kg/ha), Barmana (918.4 kg/ha) and Dharamshala (1366.4
kg/ha). Plants lacking in potassium do not have enough en-
ergy to properly grow, their roots are not well developed
and they have weak stems. Potassium deficient plants can
not regulate and use water efficiently.
Excess potassium in plants can reduce the potential pro-
duction of the crop inducing deficiencies of other nutrients
like calcium and magnesium.
Zinc is an essential component of various enzyme sys-
tems for energy production, protein synthesis and growth
regulation. Its values in the study area varied from 4.3 ppm
in Sundernagar of Kangra district to 0.4 ppm in Barmana of
Bilaspur district. The critical level for zinc in soils is 0.6
ppm. But Barmana of District Bilaspur has lower value of
zinc (0.4 ppm) than the permissible limit, while some places
reported zinc content higher than the permissible limit like
Nohangi (0.62 ppm), Ratti (0.68 ppm), Yol (0.69 ppm),
Sujanpur Tira (0.7 ppm), Bhota (0.80 ppm), Kularu and
Dangar (1.3 ppm), Dangar (2.2 ppm), Lambloo (2.33 ppm),
Sandhol (2.47 ppm), Kangra (3.48 ppm) and Sundernagar
(4.3 ppm). Zinc deficient plants exhibit delayed maturity.
The most visible zinc deficiency symptoms are short
internodes and decrease in leaf size. Zinc deficiency is mainly
found in sandy soils. Excessive uptake of zinc by plants
causes stunting of short, curling and rolling of young leaves,
death of leaf tops, and chlorosis.
Copper is necessary for carbohydrates and nitrogen me-
tabolism. It is also required for lignin synthesis, which is
needed for cell wall strength and prevention of wilting. Simi-
larly copper was found from 3.22 ppm to 0.1 ppm in
Sundernagar and Dangar respectively. The critical level of
copper in soil is 0.2 ppm. Some places reported copper con-
tent lower than the permissible limit, that is Dangar (0.1
1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R

ppm), and higher than the permissible limit, that is Bhota
(0.3 ppm), Dharamshala (0.53 ppm), Sandhol (0.66 ppm),
Sujanpur Tira and Barmana (0.7 ppm), Ghagas (0.78 ppm),
Ratti (0.8 ppm), Kularu (1.0 ppm), Nohangi (1.0 ppm),
Lambloo (1.20 ppm), Chamunda (1.25 ppm), Yol (1.63 ppm),
Kangra (2.13 ppm) and Barmana (3.22 ppm). Deficiency of
copper results in stunning of plants. Concentration above
20 µM prevents root growth and result in death of the trans-
planted cuttings. Copper toxicity in many nontolerant plants
is associated with the disturbance of mitosis, inhibition of
root elongation and damage to root cell membrane. Copper
also inhibits several enzymes, which play important role in
photosynthesis. Excess copper may cause iron deficiency.
The effect of copper toxicity is largely on root growth and
morphology.
Iron is involved in the production of chlorophyll. It is
also a component of many enzymes associated with energy
transfer, nitrogen reduction and fixation and lignin forma-
tion. It was established that the iron varies from 26.33 ppm
in Dharamshala to 3.5 ppm in Yol, whereas critical level of
iron in soil is 4.5 ppm. Some places reported iron content
lower than the permissible limit, that is Yol (3.5 ppm),
Dandor (3.82 ppm), Kularu (4.00 ppm), Kangra (4.05 ppm),
Sujanpur Tira (4.2 ppm), and higher than the permissible
limit, that is Ratti (4.93 ppm), Nouhangi (5.72 ppm), Bhota
(5.8 ppm), Dangar (6.25 ppm), Lambloo (7.11 ppm),
Barmana (7.6 ppm), Sundernagar (7.67 ppm), Ghagar (8.08
ppm), Chamunda (21.42 ppm) and Dharamshala (26.33
ppm). Iron deficiencies are mainly manifested in leaves due
to low level of chlorophyll. Severe iron deficiencies cause
leaves to turn completely yellow or almost white and then
brown as they die. Too much iron can affect the chlorophyll
itself, causing it to change and inhibiting the plant’s ability
to properly absorb energy form sunlight and as a result it
become difficult for plants to perform necessary biochemi-
cal reactions and prepare food.
Manganese is necessary in photosynthesis, nitrogen me-
tabolism and to form other compounds required for plants
metabolism. It was found that it varies from 0.4 ppm at Yol
of Kangra district and Nohangi to 23.0 ppm at Barmana of
District Bilaspur. The critical level of manganese in soil is
1.0 ppm. Some places reported manganese content lower than
the permissible limit, that is Yol (0.4 ppm), Nohangi (0.4
ppm), Lambloo (0.78 ppm), Sujanpur Tira (0.85 ppm),
Sundernagar (0.86 ppm), Sandhol and Ratti (0.9 ppm), Bhota
(0.92 ppm), and higher, that is Chamunda (1.21 ppm), Kangra
(1.72 ppm), Dharamshala (2.58 ppm), Ghagas (9.56 ppm),
Dandor (11.73 ppm), Kularu (13 ppm) and Barmana (23.0
ppm).
Interveinal chlorosis is a characteristic of manganese de-
 Ratan Chand Sharama and Shivani Dogra
ficiency symptom. In very severe manganese deficiency
brown necrotic spots appear on leaves, resulting in prema-
ture leave drop or delayed maturity. Manganese toxicity
symptoms include yellowing of leaves, which later darkens
into small brown spots. Manganese toxicity becomes prob-
lem in soil with manganese containing minerals.
$&.12:/('*(0(17
The authors are thankful to the Department of Agriculture
and Deputy Director, Agriculture, Hamirpur for the
laboratory facilities.
5()(5(1&(6
Alloway, B. I. 1990. Heavy Metals in Soils. Blackie, Glasgow and
London.
Arey, N.C. and Jagetia, B.L. 1997. Relative toxicity of cadmium, lead and
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soil phosphates. Zesz Prab. Nauk Rol., 492: 241, (in Polish).
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Priya K. L., Gabriela Jennifer, G. Lizia Thankam, Sophia Abraham Thankam and Mariam Mathew
School of Civil Engineering, Karunya University, Coimbatore-641 114, Tamil Nadu, India

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,1752'8&7,21
Coimbatore, the textile capital of south India, is a highly
populated industrial city, situated in the western Tamil Nadu,
near the State of Kerala. It is located between 10°55’-11°10’N
latitude and 77°10’-76°50’E longitude at an altitude of 470
m. It is surrounded by mountains on the west, with reserve
forests and the Nilgiri Biosphere Reserve on the northern
side. It is situated on the banks of River Noyyal, which forms
a sub-basin of Cauvery river. The Noyyal is a seasonal river
having good flow only for short periods during the north-
east and south-west monsoon (Paul & Prakash Nelliyat 2007).
The Noyyal river emerges in the Vellingiri hills of Western
Ghats and flows over a distance of 180 km in an area of
3510 sq. km to join the River Cauvery at Karur district (Priya
et al. 2008). There are many industries like textile mills, foun-
dries and manufacturing industries in the city. But the city
lacks proper drainage system and the facilities for treatment
of industrial, municipal, domestic and hospital wastes are
poor. Till now there is no integrated sewerage system. The
existing drainage and sewerage systems are of open type,
discharging wastes into lakes, wetlands and the Noyyal river
without appropriate treatment. The city is rich with 28
wetlands, mostly fed by the River Noyyal. The river and the
river-fed wetlands support a large number of plants (Chandra
Bose & Nair 1988). The Noyyal’s canals feed the lakes, most
of them are interconnected and fill one after the other by
natural gradient. These lakes cover an area of 2000 acres,
thousands of which are within the city limit itself. The lakes
are connected in chain link manner, well planned by the an-
cient rulers. Presently, the system has been disrupted and
deteriorated due to mismanagement and overall negligence.
Most of these wetlands get dried in summer and serve as
dumping yard for garbage and industrial wastes (Mohanraj
et al. 2000). Wetlands situated in the vicinity of cities un-
dergo rapid degradation due to various factors related with
city development like waste dumping, deforestation, indus-
tries and large scale reclamation for other uses. Inflows of
wastewater carrying loads in due course of time overwhelm
natural assimilation and carrying capacity of receiving wa-
ter bodies (Prusty 2008). The bund of many wetlands is un-
scientifically used for open defecation by humans as well as
animals. The quality of wetlands is so degraded that they are
not suitable for drinking purposes. Water from some of the
wetlands is used for agricultural purposes. Many of these
wetlands support rich flora and fauna, which are reported in
earlier studies (Chandra Bose & Nair 1988, Lalitha Vijayan
et al. 2007). The flora and fauna will be greatly affected by
degradation of water quality. As the environment of bottom-
dwelling organisms is the final deposition of pollutants, they
are subjected to greater pressures (Gupta & Sharma 2005).
In addition to this, the degradation of surface water poses
 Priya K. L. et al.
more stress on groundwater and results in overexploitation
of groundwater for domestic and agricultural purposes. In
1970’s Coimbatore district was declared as a drought prone
district. In 1980’s UNDP warned that Coimbatore district
had the record of fastest depleting groundwater level in the
world. This happened due to degradation of surface water
quality. It becomes mandatory to minimize the surface wa-
ter pollution to safeguard the water resources of the city.
Government and other NGOs have taken some steps to pre-
vent further pollution of surface water. To ascertain the ex-
tent of pollution of the water bodies in the current situation,
seven wetlands which fall in the city limits, were selected
for the study.
0$7(5,$/6$1'0(7+2'6
Seven wetlands falling within the city limit namely Kuruchi
Kulam, Perur lake, Selvapuram (Selvachinthamani) lake,
Kumaraswami lake, Ukkadam lake, Narasampathi lake and
Singanallur lake were selected for the study. Of these, the
pollution intensity of Kumaraswami lake and Narasampathi
lake are poorly known from earlier studies. The study area
showing the location of wetlands is shown in Fig. 1.
Kurichikulam lake has a registered ayacut of 452 acres
in the catchment area, 6.272 sq. km of free and 12.162 sq.
km of combined area. The full lake level is 10.75m. Perur
lake is located in the village Perur. It has a registered ayacut
of 866 acres in the catchment area, 2.227sq. km of free and
5.888 sq. km of combined area. The full lake level is 14.8m.
The lake is surrounded by small villages and agricultural
lands. Selvachinthamani lake is located in the village
Selvapuram in Kumarapalayam. The capacity of the lake is
3.02 MCft. Its registered ayacut is 72 acres with a water
spread area of 37.07 acres. The catchment area is 16 sq. km.
There is only one sluice present in this lake. The full lake
level is 6m. Kumaraswamy lake is located in the village
Kumarapalayam. The capacity of the tank is 19.89 MCft. Its
registered ayacut is 190 acres with a water spread area of
93.67 acres. The catchment area is 16 sq. km. There are two
sluices present in this lake. The full lake level is 10.5m.
Ukkadam lake is located in the village Ukkadam. It has a
registered ayacut of 231 acres in the catchment area of 1.90
sq. km. The full lake level is 14.8m. It is situated near the
municipal bus stand. The area is highly urbanized and re-
ceives both municipal and industrial sewage. The dumping
of solid waste was practiced here earlier, but now the lake is
protected by a boundary wall, and dumping of solid waste is
almost stopped. But still near the road side, on the bunds,
open defecation was noticed during the sample collection.
The Narasimpathy lake is situated in village Veerapatti. The
capacity of the lake is 59.8 MCft. The lake has 560.65 acres
of registered ayacut with water spread area of 122.51 acres.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
The catchment area is 16 sq. km. There are two sluices present
in the lake and its full level is 11m. Singanallur lake has a
water spread area of 1.153 sq. km. The capacity of the lake is
1.48 Mm3. The registered irrigated area is 337.1 ha. The lake
is partly affected with eutrophication on the northern side
during sample collection. This lake was earlier used for rec-
reational boating, but due to the discharge of domestic waste,
it was affected with eutrophication, and now the boathouse
has shut off its working. Efforts have been made by NGOs
and the Government to remove the water hyacinth. The ear-
lier studies by Mohanraj et al. (2000) pointed out the status
of Singanallur wetland as affected fully by eutrophication.
Samples were collected from each lake at three different
locations in pre-cleaned bottles of 2 L capacity to get a rep-
resentative sample. The samples were transferred to the labo-
ratory in a time duration of 8 hours and analysed for various
physico-chemical parameters by standard methods
(Greenberg et al. 1992). The parameters analysed include
pH, total dissolved solids (TDS), dissolved oxygen (DO),
total alkalinity (TA), chlorides (Cl), total hardness (TH), sul-
phates (Sulp), nitrates (Nit), sodium (Na), potassium (K),
calcium (Ca) and magnesium (Mg) during the postmonsoon
(August 2009) and premonsoon season (February 2010).
The Water Quality Index (WQI) for the samples was cal-
culated from the physico-chemical parameters using the fol-
lowing expression.
WQI = antilog Wi log10 qi ...(1)
Where Wi is the weightage for each parameter. It is cal-
culated as,
Wi = K/Si ...(2)
K is the proportionality constant and Si is the standard
value for water quality parameter as per BIS/ICMR for each
parameter.
K is calculated as,
K = 1/ (1/Si ) ...(3)
The standard values for water quality parameters and their
weightage are given in Table 2. The quality rating, qi is cal-
culated as,
qi = 100 * {(Vactual – Videal)/(Vstandard – Videal)} ...(4)
Where Vactual is the observed value for each parameter,
Videal is the ideal value for each parameter and Vstandard is the
standard value for each parameter as per BIS/ICMR stand-
ards. Videal for pH is taken as 7 and that for DO is taken as
14.6 mg/L and for all other parameters, value is 0.
The Hill Piper diagram is a trilinear diagram having two
triangles, one of which shows the cations and the other shows
the anions. The cation and anion fields are combined to show
 MONITORING POLLUTION INTENSITY OF WETLANDS OF COIMBATORE 

Fig. 1: Study area showing the location of lakes.

a single point in a diamond-shaped field, from which the
characteristics of the water samples are drawn.
5(68/76$1'',6&866,21
The results of physio-chemical analysis of water are given
in Tables 1a and 1b. The pH of water samples from all the
wetlands lie within the permissible limit as per BIS stand-
ards of 6.5 to 8.5 during both postmonsoon and premonsoon.
The highest pH was found to be for Ukkadam lake during
both the seasons. A pH of 7.49 was reported in earlier stud-
ies on Ukkadam lake during the month of March 2009 by
the same author (Priya et al. 2010). The rise in pH of
Ukkadam wetland indicates increased pollution. The chlo-
rides varied from 10.6 mg/L for Perur lake to 540 mg/L for
Singanallur lake during the postmonsoon, and from 23.34
mg/L for Perur lake to 575 mg/L for Ukkadam lake during
the premonsoon. The human as well as animal defecation is
the main cause for high chlorides in Ukkadam wetland. The
chlorides during postmonsoon were found to be less as com-
pared to premonsoon due to the fact that during premonsoon,
the water level falls and as a result, there is an increase in
concentration of pollutants. The high chlorides in Singanallur
wetland during postmonsoon compared to pre monsoon is
due to land leachate along with runoff water during monsoon.
The high chlorides in Ukkadam wetland are probably due to
the joining of effluents form bleaching industries. Higher
chlorides greater than the permissible limit prescribed by
BIS standards was found for water samples from
Selvachinthamani lake and Singanallur lake. Dissolved oxy-
gen was found to vary from 2.65 mg/L to 6.26 mg/L during
the postmonsoon. During the premonsoon, dissolved oxy-
gen varied from 2.95 mg/L to 6.5 mg/L, the lowest for
Narasampathy lake, and highest for Kumaraswami lake. The
DO for all the samples except for Narasampathy wetland
was found to be appreciable for aquatic life during the
premonsoon. A slight increase in DO during the premonsoon

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Priya K. L. et al.

Fig. 2a: Seasonal variation of chlorides.

Fig. 2c: Seasonal variation of pH.

Fig. 2b: Seasonal variation of dissolved oxygen.

Fig. 2d: Seasonal variation of alkalinity.
is due to the fact that the temperature was slightly low com-
pared to the postmonsoon and that DO increases as tempera-
ture decreases. The total alkalinity ranged from 120.3 mg/L
to 630 mg/L during the postmonsoon and from 82.5 mg/L
to 855 mg/L during the premonsoon. The high alkalinity of
water samples from Ukkadam and Singanallur lakes may be
due to the carbonates which reach the lake from the domes-
tic sewage. The alkalinity of Ukkadam lake was high com-
pared to the reports of Rachna (2009) and Priya (2010). This
shows the increase in pollution intensity in the Ukkadam
wetland. The alkalinity of Singanallur lake and Kurichi lake
also showed a hike compared to the reports of Priya (2010).
Total hardness ranged from 55 mg/L to 320 mg/L, the high- Fig. 2e: Seasonal variation of total hardness.
est being for Ukkadam lake during the postmonsoon as well 90.75 mg/L and are within the permissible limit specified
as premonsoon. The total hardness for all the samples was by BIS drinking water standards. Nitrates ranged from 1
within the permissible limit during the postmonsoon whereas mg/L to 16.2 mg/L, the highest value was found to be for
for the premonsoon, it was slightly greater than the permis- Singanallur wetland. High value of nitrates more than 45
sible limit for Ukkadam lake. High value for sodium ions mg/L may lead to methaemoglobinaemia or blue baby syn-
was found for the samples from Selvapuram wetland, drome in infants. The BOD ranged from 1.8 mg/L to 12.7
Ukkadam wetland and Singanallur wetland during the mg/L during the postmonsoon and from 1.2 mg/L to 11.66
premonsoon. The chlorides of these wetlands were also found mg/L, during the premonsoon. BOD values greater than
to be high that indicates the presence of sodium chloride in 20mg/L indicate organic pollution.
the samples collected from Selvapuram lake, Ukkadam lake
and Singanallur lake. High sodium chloride in samples in- The Figs, 2.a to 2.e show the seasonal variation of water
dicate pollution from human and animal waste, which en- quality characteristics of the wetlands.
ters the wetland through open defecation practices in the vi- The water quality characteristics are slightly higher dur-
cinity of these wetlands. Sulphates ranged from 1 mg/L to ing the premonsoon as compared to the postmonsoon sea-

9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
 MONITORING POLLUTION INTENSITY OF WETLANDS OF COIMBATORE 

Table 1.a: Physio-chemical characteristics of wetlands (postmonsoon).

Parameters BIS Sampling lakes

standards

L1 L2 L3 L4 L5 L6 L7

1 Chlorides (mg/L) 250 39.3 10.6 33.46 101.1 477.2 25.04 540
2 pH 6.5-8.5 7.66 7.46 7.64 7.65 7.86 7.13 7.75
3 Dissolved Oxygen (mg/L) NA 5.48 6.26 2.65 4.75 3.47 3.38 5.0
4 Total Alkalinity (mg/L) 200 273 120.3 643 410 390 25.6 630
5 Total Hardness (mg/L) 300 206.7 85.67 131.33 123.33 220 15 188
6 Calcium (Ca2+) (mg/L) 75 3.2 2.92 6.12 2.8 5.47 2.53 56
7 Magnesium (Mg2+) (mg/L) 30 3.06 2.034 3.895 3.96 2.017 2.104 11.664
8 BOD (mg/L) NA 7.11 1.8 12.7 8.63 10.3 10.82 5.9

Table 1.b Physio-chemical characteristics of wetlands (premonsoon).

Parameters BIS Sampling lakes

Standards

L1 L2 L3 L4 L5 L6 L7

1 Chlorides (mg/l) 250 95 23.34 310 118.4 575 25.5 340

2 pH 6.5-8.5 7.47 7.2 7.34 7.65 7.84 7.49 7.73
3 Dissolved Oxygen (mg/L) NA 5.75 5.13 4.6 6.5 3.95 2.95 5.1
4 Total Alkalinity (mg/L) 200 565 139.3 800 560 855 82.5 730
5 Total Hardness (mg/L) 300 212.5 101.6 210 141.98 320 55 159.55
6 Calcium (Ca2+) (mg/L) 75 47 20.26 66 32 44 14.4 15.03
7 Magnesium (Mg2+) (mg/L) 30 23.08 12.41 10.93 15.06 51.03 4.62 29.64
8 Sodium (Na2+) (mg/L) NA 30.1 12 264.8 17.4 292.6 15.5 191
9 Potassium (K+) (mg/L) NA 4.9 4.6 10 4.9 16.4 2 19
10 Sulphates (mg/L) 200 6.5 3 69 19 33.5 1 90.75
11 Nitrates (mg/L) 45 1 9.2 5 10 6 8.8 16.2
12 BOD (mg/L) NA 6.21 5.7 8.8 1.2 9.8 11.66 5.7
Water Quality Index 51.33 41.23 53.93 59.13 82.03 52.63 72.53

Note: NA-Not specified in BIS drinking water standards

Table 2: Irrigation characteristics of wetland water samples.

Samples SAR ESR % Na MH (%) RSC EC

L1 0.929 0.319 22.95 44.7 4.4 443.05

L2 0.518 0.518 19.46 50.2 0 217.43
L3 7.95 7.95 72.1 21.5 7.79 1992
L4 0.636 0.636 20.28 43.7 5.94 863.4
L5 7.12 7.12 64.99 65.7 7.28 1997
L6 0.91 0.91 36.87 34.6 0.162 215.4
L7 6.58 6.58 69.26 76.5 8.37 1445

Note: SAR-Sodium Adsorption Ratio, ESR-Exchangeable Sodium Ratio, % Na-% Sodium, MH-Magnesium Hazard, RSC-Residual Sodium Carbonate
(meq/L), EC-Electrical Conductivity (micromho/cm)
son due to the fact that during postmonsoon the storm water
adds to the dilution of concentration of pollutants. The dis-
solved oxygen was found to be slightly higher during the
premonsoon because the water temperature was low during
the month of premonsoon compared to postmonsoon. The
Fig. 3 shows the Piper diagram for the water samples. The
Piper diagram gives an idea of the type of water of the
wetlands and indicates about the dominating pollutant in a
particular wetland. Fig. 4 shows the Schoeller diagram show-
ing the distribution of ions in the study area.
It was found that the water samples from Kurichi lake,
Kumaraswami lake and Narasampathi lake exhibited a wa-
ter type of Ca(HCO 3) 2 whereas Selvapuram lake and
Singanallur lake exhibited a water type of NaHCO3. The type
of water found in Perur lake was of Mg(HCO3)2 type and
that in Ukkadam lake was of NaCl type. As the region is not
a coastal region, the chance of occurring NaCl water type in

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 Priya K. L. et al.

Fig. 3: Piper diagram for the wetland water samples.

Fig. 4: Schoeller diagram for the study area.

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 MONITORING POLLUTION INTENSITY OF WETLANDS OF COIMBATORE 

Table 3a: Water class based on sodium adsorption ratio. the water from Kurichi wetland, Perur wetland and
Narasampathi wetland fulfil the requirements for irrigation,
Sodium SAR Water class Samples
hazard class while Selvapuram wetland, Ukkadam wetland and
Singanallur wetland are degraded in quality and the water
S1 < 10 Excellent All samples cannot be used for irrigation purposes. Based on USSL
S2 10-18 Good Nil
(United States Salinity Laboratory) classification system,
S3 18-26 Poorly suitable Nil
S4 > 26 Unsuitable Nil Perur lake and Narasampathi lake falls under S1C1, Kurichi
lake comes under S1C2 and the remaining lakes come under
Table 3b: Water class based on % sodium. S1C3 group. The registered irrigated area for Ukkadam lake
% Sodium Water Class Samples and Selvapuram lake is quite small, while that of Singanallur
lake is as large as about 350 hectares. In order that crops do
< 20 Excellent L2 not get detrimental effects, suitable management measures
20-40 Good L1, L4, L6 to reduce the pollution of these wetlands become necessary.
40-60 Permissible Nil
60-80 Poorly suitable L3, L5, L7 This should be made possible only by the combined efforts
> 80 Unsuitable Nil of Government, NGOs and the public participation.
Table 3c: Water class based on electrical conductivity. At present, the water from the wetlands is not used by
the public for drinking purposes, but it is being used for irri-
Salinity Salinity hazard Conductivity Samples gation and livestock. The wetlands also help in recharging
Hazard class (micro mho/cm) the groundwater which is another source for domestic and
C1 Low salinity, no <250 L2, L6 agricultural use. So if the quality of water in the wetlands is
detrimental effects expected not controlled, the water crisis of the city will aggravate and
C2 Medium salinity, 250-750 L1 a situation will arise when the entire surface water gets dete-
detrimental effects to riorated. The polluted water percolates and recharges the
sensitive crops
C3 High salinity, 750-2250 L3, L4. groundwater, thus, resulting in the qualitative degradation
adverseeffects on L5, L7 of groundwater also. Steps need to be implemented to mini-
many crops mize the pollution of these wetlands.
C4 Very high salinity > 2250 Nil
suitable only for salt &21&/86,216
tolerant plants
Water quality of Coimbatore wetlands has shown a deterio-
Ukkadam wetland is definitely due to the entry of human ration in the past few decades. The present study reveals that
and animal waste into the wetland. These practices have led the water quality of the wetlands has degraded in past few
to the degradation of water quality, and it becomes evident years. The water quality characteristics during the
that the water from these wetlands does not fulfil the require- premonsoon were found to be higher as compared to that of
ments of drinking water. The water from these wetlands is postmonsoon. The water types of the study area were
being used for irrigation purposes in the nearby vicinity. Ca(HCO3)2, NaHCO3, Mg(HCO3)2 and NaCl type. The
The suitability of water for irrigation purposes can be Ukkadam wetland exhibited the water type of NaCl due to
ascertained by certain parameters such as Sodium Adsorp- the fact that it continuously receives human waste as well as
tion Ratio (SAR), Exchangeable Sodium Ratio (ESR), % municipal sewage. Some of the wetlands such as Kurichi
sodium (% Na), Magnesium Hazard (MH), Electrical Con- wetland, Perur wetland and Narasampathi wetland can be
ductivity (EC) and Residual Sodium Carbonate (RSC). used for irrigation as they fulfil the requirements of irriga-
tion water. The water quality of Singanallur wetland and
SAR = Na+/[(Ca2+ + Mg2+ )\2] ½
Ukkadam wetland was so worse that it cannot be used for
ESR = Na+/(Ca2+ + Mg2+) any purpose. Since the city do not have proper sewerage sys-
% Na = Na+ × 100 / (Ca2+ + Mg2+ + Na+ + K+) tem and wastewater treatment plant, the sewage, both do-
mestic and industrial, reaches the wetlands. Therefore, it is
MH = Mg2+/(Ca2+ + Mg2+)
suggested to have a proper sewerage system for the city along
RSC = (HCO3- + CO32-) – (Ca2+ + Mg2+) with a combined sewage treatment plant which needs to be
The irrigation characteristics of water samples of the properly monitored and maintained regularly. There is a gen-
wetlands are summarized in Table 2 and the classification of eral tendency to dump solid wastes on the banks of the lakes.
waters based on suitability for irrigation is given in Table 3a This will leach down into the wetland during rain and thereby
to Table 3c. From this classification, it becomes evident that polluting the water. This will not only pollute the lake wa-

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Priya K. L. et al.
ters, but the leachates also percolate into the groundwater to
cause its pollution. So there is necessity for a well planned
collection system of solid wastes from the city to reduce
dumping of wastes on open land. The city is in dire need of
a solid waste treatment plant along with its proper collec-
tion and transportation. The hygienic conditions of the city
need to be uplifted by construction of comfort stations at
different locations in order to prevent open defecation. These
should be followed by continuous monitoring of the quality
of water so that well planned hygiene city conditions can be
attained.
5()(5(1&(6
BIS: 10500, 1991. Specifications for Drinking Water, Indian Standard In-
stitutions (Indian Bureau of Standards), New Delhi.
Chandra Bose, M. and Nair, N. C. 1988. Flora of Coimbatore. Bishen Singh
Mahendra Pal Singh, Dehra Dun.
Greenberg, A.E., Clesceri, L.S. and Eaton, A.D. 1992. Standard Methods
for the Examination of Water and Wastewater. 18th edition. American
Public Health Association, American Water Works Association and
Water Pollution Control Federation, Washington.
Gupta, K. and Sharma, A. 2005. Macroinvertebrates as indicators of pollu-
tion. Journal of Environmental Biology, 261(2): 205-211.
Lalitha Vijayan, Somasundaram, S. and Deivanayaki, M. 2007. Status of
wetland birds in Coimbatore district. Annual Report 2006-2007, Salim
Ali Centre for Ornithology and Natural History, pp. 26-28.
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
Mathew, M., Mohanraj, R., Azeez, P.A. and Pattabhi, S. 2003. Speciation
of heavy metals in bed sediments of wetlands in urban Coimbatore,
India. Bull. Environ. Contam. Toxicol., 70: 800-808.
Mohanraj, R., Sathishkumar, M., Azeez, P.A. and Sivakumar, R. 2000.
Pollution status of wetlands in urban Coimbatore, Tamilnadu, India.
Bull. Environ. Contam. Toxicol., 64: 638-643.
Paul, P. Appasamy and Prakash Nelliyat 2007. Compensating the loss of
ecosystem services due to pollution in Noyyal river basin, Tamil Nadu.
Working Paper 14/2006.
Priya, K. L,. Nalini Jebastina and Prince Arulraj, G. 2008. Qualitative zon-
ing of groundwater bodies of Coimbatore city using GIS. Proceedings
of the National Conference on RACE.
Priya, K. L., Prince Arulraj, G., Ansel Jose and Jayakumar, G. 2010. As-
sessment of pollution intensity of wetlands of Coimbatore city. Pro-
ceedings of the National Conference on Sustainable Water Resources
Management and Impact of Climate Change.
Prusty, B. A. K. 2008. Role of detritus in trace metal dynamics of a wetland
system: A case study of Keoladeo National Park, Bharatpur. Report to
Council of Scientific and Industrial Research, New Delhi, India.
Rao, K.R., Krishnan, R., Chakraborthy, A.K. and Deekshatulu, B.L. 1980.
Correlation of water quality parameters and products of Exotech radi-
ometer bands. J. Photo-lnt. & Remote Sensing, 8(2).
Senthilnathan, S. and Azeez, P.A. 1999. Water quality of effluents from
dyeing and bleaching industries in Tirupur, Tamilnadu, India. J. Ind.
Pollu. Control, 15: 77-86.
Shanthi, K., Ramasamy, K. and Lakshmanaperumalsamy, P. 2003. Sedi-
ment quality of Singanalluar wetland in Coimbatore, Tamil Nadu, In-
dia. Bull. Environ. Contam. Toxicol., 70: 372-378.
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Mazher Sulthana, R. Vasanthi and J. N. Jayaprakash
Department of Advance Zoology and Biotechnology, Unit of Human Health and Environmental Biotechnology,
Presidency College, Chennai-600 005, Tamil Nadu, India

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,1752'8&7,21
In recent times there has developed an increasing awareness
on the value of knowledge of freshwater bodies. Freshwater
ponds appear to be ideal models of biotope for studies on
microecology and microcosms (Ganapathy 1980). Manmade
lakes and ponds or impounded water body are peculiar
biotopes requiring intensive detailed investigations. Most
of the reports on pond ecosystems include the work of
Munawar (1970), Saha & Pandit (1986), Rajendran Nair
(2000) and Sedamkar & Angadi (2003).
The biodiversity of phytoplankton and zooplankton in
water bodies shows a correlation with reference to their oc-
currences and the physico-chemical characteristics. The
phytoplankton serve as producers in food chain in aquatic
ecosystems and the productivity of pond depends upon the
quality of water. The zooplankton depend upon the avail-
ability of phytoplankton and form the second trophic level
in aquatic food chains. The phytoplankton and zooplankton
provide food for fishes and therefore play an important role
in fisheries. At several places there is scarcity of freshwater.
Any kind of pollution of water will further enhance diffi-
culty in procuring it naturally. Thus, there is an urgent need
to generate information about water quality and plankton
diversity, which form the basis of aquatic production. In the
present investigation, focus has been laid on the study of
water quality in terms of physico-chemical characteristics
and plankton diversity of the Arasankulam pond, situated at
Veppampattu in Thiruvallur district of Tamil Nadu.
0$7(5,$/6$1'0(7+2'6
Study area: Arasankulam pond is located at Veppampattu
nearly 15 km away from Chennai city, which stretches be-
tween latitude 13°07’30” N and longitude 80°01’05” E. No
study has been done till date on this pond. Therefore, the
present work was undertaken for an extensive survey of
phytoplankton and zooplankton diversity in relation to
physico-chemical factors. The collected water samples were
analysed for various physico-chemical parameters by fol-
lowing the standard methods of APHA (1989). The density
of population of phytoplankton groups was estimated fort-
nightly (monthly average mean was taken), followed by the
methods of Rao (1955). Further, the algae were isolated from
these water samples and identified with help of monographs.
Zoo planktons were collected using a conical net with a
mesh size of 120 µm approximately. The samples were pre-
served with 4% formalin. The species were identified with
the help of identification keys (Sehgal 1983, APHA 1989).
The quantitative estimation was carried out by Sedgwick
Rafter cell method.
5(68/76$1'',6&866,21
Physico-chemical environment has profound influence on
biotic components in aquatic ecosystems. It controls
biodiversity, biomass and distribution of biotic communi-
ties. The physical and chemical parameters exert their influ-
ence both individually and collectively and their interactions
produce a biotic environment which ultimately conditions
 Mazher Sulthana et al.
the origin, development and finally succession of biotic
communities.
The analysis of physico-chemical characteristics (Table
1) reveal that the water and atmospheric temperature ranged
from 24-32°C and 29-36°C, whereas the turbidity varied
from 0.6-10.58 NTU. pH values showed a variation from
7.2-8.5. The total alkalinity ranged from 92-126 mg/L. As
the pond water is alkaline, hence favourable for the growth
of the fishes.
Dissolved oxygen (DO) is another vital parameter regu-
lating survival of aquatic life. It was found to be maximum
in the month of January 2009 i.e., 7.8 mg/L, and minimum
in March 2009, as 3.5 mg/L. It shows inverse relation with
the water temperature. The maximum chloride concentra-
tion was recorded to be 55 mg/L in May, while the mini-
mum of 32 mg/L during January 2009. Total hardness ranged
between 96 mg/L and 138 mg/L, which is within the per-
missible limit of drinking water standard (BIS 1991). As the
pond water is not hard, it can be used for all domestic
purposes.
The nutrient contents like nitrate, phosphate, nitrite and
silicate showed variation from 0.26 to 5 mg/L, 0.36 to 0.95
mg/L, 0.003 to 0.14 mg/L and 19.32 to 55.74 mg/L respec-
tively. Iron ranged from 0.02 to 0.29 ppm, and did not ex-
ceed the permissible limit. There was absence of manganese
in the water samples throughout the period of study.
It is generally believed that temperature is one of the im-
portant factors in aquatic ecosystems. Chako &
Krishnamoorthy (1954) could not establish any correlation
between water temperature and plankton variation. In the
present study the total alkalinity increased in summer and
decreased in monsoon period. This supports the observations
of Munawar (1970), Saha & Pandit (1986) and Rajendra Nair
(2000) that the accumulation of bicarbonate in summer is
due to increased rate of decomposition. They have discussed
the interrelationship among pH, carbon dioxide, alkalinity,
calcium, carbonate and bicarbonate. This work confirms that
the similar generalization is true for the Arasankulam pond
under present study.
Hardness was found to be maximum in summer, and
minimum in monsoon (Rajendran Nair 2000) and dissolved
oxygen being minimum in summers and maximum in win-
ters. Dissolved oxygen showed an inverse relationship with
water temperature, which is probably due to two reasons, in
summers at high temperature rate of oxidation of organic
matter in water increase and oxygen is consumed in this proc-
ess. Secondly, at higher temperature the water has a lesser
oxygen holding capacity and surplus oxygen is lost to the
atmosphere (Munawar 1970, Saha & Pandit 1986 and
Rajendran Nair 2000).
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Higher chloride content is considered as an index of water
pollution. Munawar (1970), Saha & Pandit (1986) have re-
ported lower chloride values in unpolluted pond. In the
present study, chloride range was within permissible limit.
It showed the positive correlation with water temperature,
turbidity, hardness, alkalinity, conductivity and negative
correlation with pH and dissolved oxygen. The phosphate
content being higher in monsoon and post monsoon, is prob-
ably due to the influx through rain water (Munawar 1970).
This showed positive correlation with pH, dissolved oxy-
gen and negative correlation with water temperature, hard-
ness, alkalinity and chloride. Similar findings were reported
by (Rajendran Nair 2000).
Nitrate, ammonia, silicate and sulphate content of the
Arasankulam pond was found to be higher during monsoon
period and lower in summer; similar seasonal trends has been
reported by Munawar (1970). A comparison of physico-
chemical characteristics in Arasankulam pond showed that
values for most parameters were below permissible limit
except total alkalinity.
The seasonal variations in water quality characteristics
of the pond have a marked influence on the numerical abun-
dance of plankton. Relation between these parameters and
plankton diversity has been studied by Dutta et al. (1954),
Meshram & Dhande (2000) and Mazher Sultana & Sharief
(2004). The present analysis showed the abundance of
phytoplankton as compared to zooplankton (Tables 2 & 3).
In the present study 33 species of phytoplankton and 15
species of zooplankton have been recorded. Of the total
plankton, 15 belong to Chlorophyceae (Scenedesmus
dimorphus, S. quadricauda, Pediastrum duplex, Oocystis
gigas, Spirogyra sp., Chlamydomonas sp., Ankistrodesmus
sp., Pandorina sp., Desmid sp., Oedogonium sp., Zygnema
sp., Chlorococcales sp., Volvox sp.), 9 to Cyanophyceae
(Microcystis aeruginosa, Anabaena sp., Oscillatoria sp.,
Chroococcus minutus, Lyngbya sp., Nostoc sp., Spirulina
sp., Merismopedia sp.), 7 to Bacillariophyceae (Navicula sp.,
Gomphonema sp., Cyclotella sp., Synedra sp., Fragillaria
sp., Nitzchia sp.), 2 to Euglenophyceae (Euglena sp., Phacus
sp.), 5 to Cladocerans (Daphnia sp., Ceriodaphnia sp., Moina
sp., Alona sp., Daphnia nauplius sp.), 4 to Copepods
(Thermocyclops sp., Mesocyclops sp., Diaptomus sp., Cy-
clops sp.), 4 to Rotifers (Keratella sp., Brachionus sp.,
Asplanchna sp.) and 2 to Ostracods (Cypris sp., Stenocypris
sp.) (Tables 2 and 3). In the present study, the annual perio-
dicity shows that Chlorophyceae dominates and constitutes
33% of the total phytoplankton population, followed by
Bacillariophyceae (29%), Cyanophyceae (23%) and
Euglenophyceae (14%). Maximum density of phytoplankton
was recorded in summer, and minimum in winter.
 HYDROLOGY AND PLANKTON IN ARASANKULAM POND 

Table 1: Physico-chemical characteristics of Arsankulum pond at Veepampattu from January-December 2009.

No. Parameters Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

1 Atm. Temperature 29 32 34 35 36 35 33 31 27.5 28 26 26

2 Water Temperature 26 28 30 30 32 29 29 29 25.5 26 24 24
3 Turbidity 4.4 4.6 0.9 7 0.6 0.6 9.6 10.6 11.2 12 13.5 12.5
4 Conductivity 361 444 488 520 545 552 569 585 600 612 650 660
5 pH 7.79 8.03 8.54 7.32 7.29 8.01 8.11 8.5 8.86 9.2 8.84 8.8
6 Alkalinity 96 116 126 134.5 108 110 92 122 114.5 110.6 108 110
7 Total hardness 96 115 130 138 128 130 110 108 114 118.5 128.5 116.6
8 DO 7.8 5.35 3.5 3.8 5.05 5.56 5.82 6.58 6.93 6.59 7.04 7.53
9 Chloride 32 38 42 51 55 52 42 44 48 52 54 55
10 Nitrate 2 4 0.26 4 3 5 2 4 4.3 5.5 3.2 2.5
11 Nitrite 0.07 0.02 0.003 0.09 0.12 0.14 Nil 0.05 0.05 0.03 0.04 0.04
12 Phosphate 0.47 0.64 0.47 0.36 0.92 0.95 0.7 0.93 093 0.94 0.95 0.95
13 Iron 0.15 0.21 0.02 0.11 0.16 0.25 0.28 0.27 0.28 0.29 0.29 0.28
14 Silica 19.32 53.65 20.32 55.74 33.08 43.52 23.7 30.25 45.05 56.8 58.04 59.55
15 BOD 4 7 10 12 8 6 4 8 15.6 14.09 16.05 10.08
16 COD 16.35 25.12 28.2 37.65 2.54 7.6 10.6 15.20 16.78 20.53 22.30 20.08

The values are in mg/L except pH, temperature (ºC), turbidity (NTU) and conductivity (micromhos/cm).

Table 2: Monthly occurrence of phytoplankton in Arasankulum pond from January-December 2009.

Period Cyanophyceae Chlorophyceae Bacillariophyceae Euglenophyceae Total

Jan 10 18 16 27 71
Feb 20 28 21 25 94
Mar 30 40 25 16 111
Apr 35 45 30 16 126
May 26 45 30 10 111
Jun 24 35 23 12 94
Jul 12 36 12 07 67
Aug 20 15 10 08 53
Sep 20 12 31 02 65
Oct 18 10 23 03 54
Nov 16 08 22 01 47
Dec 08 06 20 02 36
Total 209 298 263 129 893
Percentage 23.40% 33.37% 29.45% 14.44%

Table 3: Monthly occurrence of zooplankton in Arasankulum pond from January-December 2009).

Period Cladocera Ostracoda Copepoda Rotifera Total

Jan 26 10 15 16 67
Feb 28 12 17 12 69
Mar 15 08 28 28 79
Apr 28 10 25 30 77
May 12 10 28 25 63
Jun 13 15 18 22 87
Jul 10 15 20 20 65
Aug 26 14 18 18 80
Sep 15 15 12 08 50
Oct 07 10 08 10 35
Nov 10 5 14 12 41
Dec 10 8 16 10 49
Total 202 132 217 211 762
Percentage 26.50% 17.32% 28.47% 27.69%

1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\x9RO1R
 Mazher Sulthana et al.
Zooplankton in water of Arasankulam pond belong to
four main taxonomic groups, Copepoda, Rotifera, Cladocera
and Ostracoda. The knowledge of their abundance, species
diversity and spacial distribution is important in understand-
ing profodynamics and trophic progression of water bodies
as pointed out by Mathew (1977), Verma & Dutta Munshi
(1987) and Pulle & Khan (2003).
The monthly average and total number of copepods var-
ied from 8 to 28 per litre during the year 2009. Copepod
population was found to be higher in the summer, while lower
in monsoon and winter. Water temperature and availability
of food organisms affect the Copepoda population.
The monthly average of total number of Cladocera was
observed to vary from 7 to 28 per litre during the study pe-
riod. They were maximum in summer and winter, and mini-
mum in premonsoon. The maximum population of Cladocera
in winter may be attributed to favourable temperature and
availability of abundant food in the form of bacteria,
nanoplankton and suspended detritus (Baker 1979). During
summer and monsoon, the factors like water temperature,
dissolved oxygen, turbidity, and transparency play an im-
portant role in controlling the diversity and density of
Cladocera. The monthly average of total number of rotifers
varied from 8 to 30 per litre. Rotifer populations are very
useful in indicating the water quality, particularly in pollu-
tion studies (Mazher Sultana & Sharief 2004). Chandrasekhar
(1996) observed that in summer and monsoon, the factors
like water temperature, turbidity, transparency and dissolve
oxygen play an important role in controlling the diversity
and density of rotifers. In the present study, the rotifer number
was found maximum in summer, and minimum in winter,
which corroborates with the work of Pulle & Khan (2003).
The ostracod population in the pond was higher in
monsoon and lower in winter and summer. The higher popu-
lation of ostracod during monsoon may be due to the abun-
dance of fine detritus to which omnivorous organisms switch
over during monsoon from their natural benthic habitat and
use bacteria, moulds and algae as food (Tonapi 1980). Thus,
9RO1Rx1DWXUH(QYLURQPHQWDQG3ROOXWLRQ7HFKQRORJ\
it may be concluded that the density of plankton is depend-
ent on different abiotic factors either directly or indirectly.
5()(5(1&(6
APHA 1989. Standard Methods for the Examination of Water and
Wastewater. American Public Health Association, Washington, D C.
17th edition.
Baker, R.L. 1979. Specific status of Keratella cochlearis (Gosse) and K.
earlinare Ahlastrom (Rotifera: Brachionidae): Morphological and eco-
logical consideration. Can. J. Zool., 57(9): 1719-1722.
Chacko, P.I. and Krishnamoorthy, J.B. 1954. On the plankton of three fresh-
water fish ponds in Madras city. Ind. Symp. Mar. Freshwater Plank-
ton, Indo-Pacific Fish Council, UNESCO.
Dutta, N., Malhotra, J.C. and Bose, B.B. 1954. Hydrology and seasonal
fluctuation of the plankton in Hooghly estuary. In: IPFC, Proc. 35-47.
Eshwarlal Sedamkar and Angadi, S.B. 2003. Physicochemical parameters
of two freshwater bodies of Gulbarga-India with special reference to
phytoplankton. Poll. Res., 22(3): 411-422.
Ganapathi, S.V. 1980. An Ecological study of garden pond containing abun-
dant zooplankton. Proc. Ind. Acad. Sci., 17: 48-58.
Mathew, P.M. 1977. Studies on the zooplankton of a tropical lake. Proc.
Symp. Warm. Water Zool. Pl. Spt. Pupl. (UNESCO). Nio, pp.
297-308.
Mazher Sultana and Dawood Sharief 2004. Water pollution studies in the
Double lake (Erettai Eri) with relation to phytoplankton. J. Aqua. Biol.,
19(1): 15-18.
Meshram, C.B. and Dhande, R.R. 2000. Algal diversity with respect to
pollution status of Wadali lake, Amaravathi, Mahatashtra. India. J.
Aqua. Biol., 15(1&2): 1-5.
Munawar, M. 1970. Limnological studies in freshwater ponds of Hyderabad,
India. Hydrobiologia, 31, 101-128.
Pulle, J.S. and Khan, A.M. 2003. Studies on zooplanktonic community of
Isapur Dam water, India. Poll Res., 22(3): 451-455.
Rajendran Nair, M.S. 2000. Seasonal variation of physico-chemical fac-
tors and their impact on the ecology of village pond at Imalia (Vidisha).
J. Ecobiol., 12(1): 21-27.
Rao, C.B. 1955. On the distribution of algae in a group of six small ponds.
J. Ecol., 41: 62-71.
Saha, L.C. and Pandit, B. 1986. Comparative limnology of Bhaalpur ponds.
Comp. Physiol. Ecol., 11(4): 213-216.
Sehgal, K.L. 1983. Planktonic Copepods of Freshwater Ecosystem. Envi-
ronment Science Series. Interprint, Mehta House, Nariana I, New Delhi,
pp. 125.
Tonapi, G.T. 1980. Freshwater Animals of India (An Ecological Approach).
Oxford and IBH publishing Co., New Delhi, pp. 341.
Verma, P.K. and Datta Munshi, J.S. 1987. Plankton community structure
of Baduka Reservoir of Bhaalpur (Bihar). Trop. Ecol., 28: 200-207.
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L. Jose and Chethan Kumar
Department of Botany, St. Albert’s College, Ernakulam, Cochin-682 018, Kerala, India

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,1752'8&7,21 members of four groups viz, Cyanophyceae, Chlorophyceae,

Kerala is a state blessed with a large number of freshwater
resources, which serve as source of water for drinking and
recreational purposes. Many of the freshwater resources are
currently under threat due to human interference. The main
reasons for this include urbanization and industrialization.
Land filling is yet another problem leading to destruction or
impairment of traditional ponds. In India many studies have
been carried out in lotic and lentic systems, but significant
study has not been carried out in the ponds of Kerala except
a few studies by Jose & Sreekumar (2005, 2006), Jose et al.
(2008). The present study is an attempt to evaluate the pol-
lution by using biological and chemical methods.
0$7(5,$/6$1'0(7+2'6
The present study was carried out in four temple ponds lo-
cated in Mattancherry Taluk of Ernakulam district. These
ponds are Dharma Sastha temple pond, Anavathil temple
pond, Tirumal Devaswom temple pond and Janardhana
temple pond.
Studies were carried out from February 2006 to January
2007. For qualitative estimation, a portion of samples was
preserved in 4% formalin and later observed under a light
microscope. The algae were identified with the help of rel-
evant monographs, books and research publications. Physico-
chemical analysis of water was carried out by standard meth-
ods (APHA 1975). For rating the water sample as high or
low organically polluted the Algal Generic Pollution Index
of Palmer (1969) was employed.
5(68/76$1'',6&866,21
In the present study the algal flora was represented by
Euglenophyceae and Bacillariophyceae. The distribution
pattern of phytoplankton in different ponds is summarized
in Table 1.
Thirty one algal genera represented the flora. Among
them 8 belonged to Cyanophyceae, 14 to Chlorophyceae,
2 to Euglenophyceae and 7 to Bacillariophyceae. The
Chlorococcales like Ankistrodesmus, Dictyosphaerium and
Scenedesmus were well distributed in all the ponds studied.
Cyanophyceae members present in all the four ponds include
Chroococcus and Oscillatoria.
The Palmer’s Algal Pollution Index is employed in all
the ponds and the results are given in Table 2. All the ponds
showed probable high organic pollution
The physico-chemical analysis of the four ponds is given
in Table 3, which represents the average value of various
parameters during the year. From the study, it was observed
that the concentration of nitrates and phosphates are consid-
erably high. The freshwater bodies receive a load of nutri-
ents from surrounding lands as well as from anthropogenic
activities, bathing and washing. Sometimes, human and cat-
tle excrement also play a significant role in increasing the
nutrient concentration of these water bodies which in turn
manifest in the phenomenon like eutrophication. Algae, be-
ing a primary inhabitant of water, play a significant role in
the ecology of these water bodies. Algal communities domi-
nated by Chrococcus, Microcystis, Ankistrodesmus,
Dictyosphaerium, Pediastrum, Scenedesmus, Trachelomonas,
Melosira and Nitzschia were found in the ponds. Their pres-
ence is an indication of organic pollution of the water as
established by Palmer (1969), Robert et al. (1974) and
Hosmani & Bharati (1980). The genus, Scenedesmus is
 L.Jose and Chethan Kumar

Table 1: Algal distribution in four ponds. Table 2: Palmer’s algal pollution index values in four ponds.

Algal Genera Pond 1 Pond 2 Pond 3 Pond 4 Algal genera Index value Pond 1 Pond 2 Pond 3 Pond 4
Anabaena + + - -
Anabaenopsis - + + - Anacystis 1 1 1 1 -
Chrococcus + + + + Oscillatoria 4 4 4 4 4
Merimospedia + + + - Phormidium 1 - 1 - -
Microcystis + + + - Chlamydomonas 4 - - - -
Oscillatoria + + + + Pandorina 1 1 - - -
Phormidium + + - - Scenedesmus 4 4 4 4 4
Raphidiopsis + - - - Micratinium 1 - - - -
Chlamydomonas + + - - Ankistrodesmus 2 - 2 2 2
Pandorina + - - - Chlorella 3 - - - -
Ankistrodesmus + + + + Closterium 1 - - - -
Coelastrum + + - + Stigeoclonium 2 - - - 2
Crucigenia + + - + Cyclotella 1 1 1 - 1
Cosmarium - + - - Melosira 1 1 1 - -
Dictyosphaerium + + - - Gomphonema 1 - - - -
Dimorphococcus - - - + Navicula 3 - - - -
Scenedesmus + + + + Nitzschia 3 3 - 3 3
Tetrastrum + - - - Synedra 2 - - 5 -
Tetraedron + + - + Euglena 5 - 5 - -
Pediastrum + + + + Phacus 2 - - - -
Oedogonium - - - + Lepocinclis 1 - - - -
Staurastrum - + - - Total 15 19 19 16
Trachelomonas + + - - According to Palmer’s Algal Pollution Index values between 0-10 indicate
Euglena - + + - lack of organic pollution, 10-15 moderate pollution, 15-20 probable high
Achnanthes + - - - organic pollution and 20 and above as confirmed high organic pollution.
Cyclotella + + - +
Cymbella + + - - Table 3: Physico-chemical analysis of waters.
Melosira + + - +
Nitzschia + + + + Parameters Pond 1 Pond 2 Pond 3 Pond 4
Synedra + - - -
Pinnularia - + - + Temperature,°C 29.5 29.6 29.8 29.7
pH 7.2 7.5 7.3 7.7
present in all the ponds, its occurrence in polluted water, Dissolved Oxygen, mg/L 5.3 5 4.8 4.4
Nitrate, mg/L 0.8 0.75 0.66 0.74
especially in eutrophicated water, is established by Tripathi
Phosphate, mg/L 0.3 0.4 0.32 0.4
et al. (1987). Palmer (1969) also gave a high ranking for this Sulphate, mg/L 0.5 0.5 0.46 0.9
genus. Microcystis (Anacystis) was considered by Singh Hardness, mg/L 37 37.6 44 45
(1973) as the best single indicator of pollution. All the ponds Chloride, mg/L 23 40.6 33.3 25.3
except the fourth pond showed the presence of Microcystis
indicating the deteriorated quality of water. Ernakulam. STARS Int. Journal, 6(2): 36-39.
Jose, L. and Sreekumar, S. Menon 2006. A study in organic pollution based
All the ponds showed probable high level of organic on algal distribution in some rural and temple ponds of Ernakulam,
pollution according to the Palmer’s index. This is substanti- Kerala, India. Nature Environment and Pollution Technology 5(2):
283-286.
ated by the physico-chemical analysis of the water, which
Jose, L., Sanjo Cine Mathew and Sreekumar, S. Menon 2008. Studies on
showed moderate levels of pollution. The study concludes organic pollution based on physicochemical and phycological char-
that the traditional water bodies in the study sites are in po- acteristics of some temple ponds of Ernakulam, Kerala, India. Nature
tential danger of being polluted by human activity. Environment and Pollution Technology 7(1): 97-100.
Palmer, C.M. 1969. A composite rating of algae tolerating organic pollu-
5()(5(1&(6 tion. J. Phycol., 5: 78-82.
Robert, D.S., Robert, W.H. and Everett, L.G. 1974. Phytoplankton distri-
APHA 1985. Standard Methods for the Examination of Water and bution and water quality indices for Lake Mead (Colorado River). J.
Wastewater. 16th Ed., American Public Health Association, Washing- Phycol., 10: 323-331.
ton, D.C. Singh, R.N. 1973. Limnological relations of Indian land waters with spe-
Hosmani, S.P. and Bharati, S.G. 1980. Algae as indicators of organic pol- cial reference to water blooms. Verh. Int.Verein. Theor. Anen. Limnol.,
lution. Phykos, 19(1): 23-26. 12: 831-836.
Jose, L. and Sreekumar, S. Menon 2005. A study on phytoplankton consti- Tripathi, A.K., Pandey, S.N. and Tiwari, R.K. 1987. Eutrophication study
tution and organic pollution in some rural and temple ponds of of Kalyanpur Pond, India. Proc. Nat. Acad. Sci. India, 57(B): 111.

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D. N. Khairnar, A. S. Kelhe and A. B. Khairnar
Mycology and Plant Pathology Research Laboratory, K.A.A.N.M.S.Arts, Commerce & Science College Satana-423 301,
Maharashtra

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,1752'8&7,21 To study the effect of fungal metabolites on seed

Seed-borne fungi are chiefly responsible for the deteriora-
tion of seeds in storage, thus reducing their viability. Seed
germination and seedling vigour are greatly influenced by
seed-borne fungi (Christensen & Kaufman 1965,
Vidhysekaran & Parsmbaramani 1973). Effect of fungal
metabolites on seed germination and seedling growth on
some wheat varieties has been studied by Thakur (1983) and
Khairnar (1987). The present work deals with the fungal di-
versity and mycotoxin effect of fungal metabolites on seed-
borne fungi and germination of seeds of some cereals.
Metabolites of Aspergillus carbonarius, A. flavus, Fusarium
oxysporum and F. moniliformae were used as they domi-
nant seed-borne fungi of some cereals like wheat, sorghum,
pear millet, finger millet, maize and rice. Tweny seven fun-
gal species have been isolated from stored seeds but only
four fungi were used in the present study.
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Seed samples of different cereals were collected in three ran-
dom samples (half kg each) from various storehouses and
markets. Standard blotter and agar plate method with
Wakman’s acid agar and Rose Bengal agar medium was used
as recommended by ISTA (1960) for isolation of seed-borne
fungi of six cereals. A. carbonarius, A. flavus, Fusarium
moniliformae and F. oxysporum were grown separately in
250 mL conical flasks each containing 150 mL sterilized
Czapek’s liquid medium and incubated for 8 days at 28° r
5°C. The cultures (mycotoxins) were filtered through
Whatman No. 44 filter paper.
mycoflora, 100 seeds of each of the test cereals were soaked
in the fugal metabolites for 24 h. Seeds soaked in sterilized
distilled water served as control. The treated seeds were
placed in each dish and 10 replicate dishes each were kept
for six cereals. After seven days of incubation at 28° r 5°C
mycoflora was observed. The effect of metabolites on seed
germination was studied by soaking seeds in the fungal
metabolites for 24 h. The seeds were sterilized in 0.1 % HgCl2
solution. The treated seeds were placed in sterile Petri plates
containing sterile moist blotting papers. Five mL of the fil-
trate was added to each Petri dish over the blotting papers
for adequate supply of the fungal metabolites. Seeds treated
with sterile distilled water and sterile Czapek’s liquid me-
dium under the same conditions served as control. The
number of seeds germinated after five days was observed
and percent germination was recorded.
To study the efficacy of metabolites on shoot and root
elongation, seeds were allowed to germinate in water for 3
days. Ten such seeds of ach cereal were placed on sterile
Petri dish over blotting paper moistened with 5 mL culture
filtrates of A. carbonarius, A. flavus, Fusarium moniliformae
and F. oxysporum separately. Blotting papers moistened with
sterile distilled water, Czapek’s liquid medium, sterile
Waksman’s medium and sterile glucose nitrate medium
served as controls. The shoot and root lengths of germinated
seedlings were measured after four days of incubation.
5(68/76$1'',6&866,21
Twenty seven fungal species were isolated from the seeds of
 D. N. Khairnar et al.

Table 1: Effect of fungal metabolites of A. carbonarius, A. flavus, F. moniliformae and F. oxysporum on seed germination (%) in different cereals.

Cereals Control CZ CW CG A. carb A. flav F. moni F. oxy

Wheat 100 70 90 80 40 00 00 00
Sorghum 70 60 70 70 30 00 00 00
Pearl millet 80 80 75 80 40 00 00 00
Maize 100 70 90 100 50 00 00 00
Rice 70 70 80 70 50 30 00 00
Finger millet 80 70 80 100 40 00 00 00

CZ-control sterile Czapek’s liquid medium, CW-control sterile Waksman’s solid medium, CG- control sterile glucose nitrate medium

Table 2: Effect of fungal metabolites of A. carbonarius, A. flavus, F. moniliformae and F. oxysporum on shoot and root elongation in different cereals.

Cereals Shoot/Root length (cm)

Cont CZ CW CG A.carb. A.flavus F.moni. F.oxysp.

Wheat 5/8 5/5 4/6 4/6 4/4 0/0 0/0 0/0

Sorghum 4/9 3/8 4/6 4/6 4/5 0/0 0/0 0/0
Pearl millet 4/9 3/7 3/7 4/7 2/4 0/0 0/0 0/0
Maize 2/7 1/5 2/4 1/6 1/4 0/0 0/0 0/0
Rice 3/8 3/6 3/7 3/8 0/6 0/0 0/0 0/0
Finger millet 4/9 3/5 3/7 4/6 3/6 0/0 0/0 0/0

CZ-control sterile liquid medium, CG- control sterile glucose nitrate medium, CW- control sterile Waksman’s medium

six cereals by blotter as well as agar plate method.
The fungal metabolites played significant role in reduc-
ing the number and population of seed mycoflora. Further,
metabolites of A. flavus F. moniliformae and F. oxysporum
were more effective than those of A. carbonarius.
Table 1 indicates that the fungal metabolites of all fungi
reduced considerable seed germination in all cereals, except
rice. Germination was suppressed by the presence of
inbibitory substances in the fungal culture filtrate and the
secretion of some mycotoxins caused seed rotting. Percent
germination increased in control, seeds treated with sterile
distilled water, and it was followed by seeds treated with
four different liquid media.
Table 2 reveals that the metabolites of A. flavus, F.
moniliformae and F. oxysporum were more inhibitory show-
ing no shoot and root elongation in all cereals expect rice.
Maximum elongation in shoot and root length was observed
in control seeds. The reduction in seed germination and sup-
pression of seedling vigour might be due to the presence of
some inhibitory substances in the fungal culture filtrates and
the secretion of some phytotoxic ingredients (mycotoxins)
in growth medium as reported by Lalithakumari et al. (1974)
in castor seeds, Tripathi (1974) in sorghum seeds and
Khairnar (1987) in pearl millet seeds.
$&.12:/('*(0(17
The authors are thankful to UGC-New Delhi for financial
support and grateful to the Principal of this college for labo-
ratory facilities.
5()(5(1&(6
Christensen, C.M.and H.H Kaufman 1965. Deterioration of stored grains
by fungi. Annual Rev. Phytopath., 3: 69-84.
Khairnar, D.N. 1987. Studies on seed-borne fungi of bajra. Ph.D. Thesis,
Marathwada University, Aurangabad.
Lalithakumari, D. and Govindaswami, C.V. 1971. Effect of seed-borne fungi
on the physicochemical properties of groundnut oil. Indian Phytopath.,
24: 283-289.
Thakur M.K. and Prasad, R.B. 1983. Effect of fungal metabolites on seed
germination and seedling vigour of Triticum vulgare var. shera.
Geobios, 10: 91-92.
Tripathi, R.K. 1974. Head fungi of sorghum, phytotoxins and their effects
on seed germination. Indian Phytopath., 27: 499-501.
Vidhyasekaran, P. and Parsmbaramani, P.C. 1973. Role of pectolytic en-
zymes in pathogenesis of obligate and facultative parasites causing
sorghum diseases. Indian Phytopath., 26: 197-209.

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Siddiqui Tasneem
Department of Zoology, Yeshwant Mahavidyalaya, Nanded-431 602, Maharashtra, India

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B arul reservoir is located in Warawant village at a dis

tance of 10 miles from Kandhar town in Nanded dis-
trict. Dam is located at latitude 18°50’ north and longitude
77°19’ east. Ichthyology is more popular branch of natural
history. Any study on aquatic systems cannot be visualized
as complete without the study of fishes as they often lie at
end of a complex food chains. Besides, forming the staple
diet for human beings, fishes are also a useful parameter of
the purity of water (Peter 1987). Seth et al. (1967) suggested
that the fishes can be utilized in monitoring water quality
for the toxic constituents that may enter the water through
the discharge of wastewaters.
Fishes from four sampling stations were collected every
month throughout the study period with the help of fisher-
men. The colour pattern of fishes was recorded at the time of
collection. The fishes were preserved in 5% formalin and
identified following the key (Fin formula) of Jhingran (1991),
Day (1994) and Jayaram (1998). The knowledge of the oc-
currence of fish in India dates back to 3 million B.C. (Hora
1956). As regards the total number of different species of
fishes, the entire world has about 21,000 species of fishes.
In India more than 1600 species of fishes have been found,
although Day (1994) has recorded 1300.
Barul reservoir has a very rich fish fauna. During the
study period fish fauna was represented by 24 fish species
belonging to 11 Families and 4 different Orders as given in
Table 1. The 4 major species of the carps were Catla, Rohu,
Mrigal and Cyprinus. The carnivorous or predatory fishes
include Cat fishes, herbivores include Rohu, Grass carp, and
Omnivores include Mrigala and Clarias. A variety of carps
belong to the Order Cypriniformes including the species of
Punctius, Cirrhinus, Cyprinus, Katia, Labeo, Barbus and
Rasbora. Eel fishes belonging to the Order Acanthopterygii
included the species of Channa and Mastacembelus. Cat-
fishes from the reservoir were of the Order Siluriformes in-
cluding species of Wallago, Mystus and Clarias. Total 24
species of fishes were found in the reservoir. The reservoir
is highly exploited for harvesting and marketing the fishes
on daily basis. The percentage of fishes in number by catch
showed a great variation and the study confirms fishes from
the order Cypriniformes dominated in the dam. Deep zone
is dominated by major carps and Cat fishes, which might be
due to the population dynamics of zooplanktons including
Rotifers. Job et al. (1955) recorded 86 species from Hirakund
dam, Lohar et al. (2003) reported 24 species from River Tapi
belonging to 7 Families and 16 Genera. The fishes of Yeldari
reservoir have been studied by Sakhare (1999). Suresh (2003)
reported 54 fish species in Loktak lake, Manipur and 15 spe-
cies were commercially important. About 43 species were
present in which 18 were commercially important. Sakhre &
Joshi (2003) reported the Ichthyofauna of Bori reservoir in
Maharashtra with a total of 20 species of fishes belonging to
14 Genera falling under 4 Orders (Cypriniformes, Perciformes,
Siluriformes, Osteoglossiibrmes). Cypriniformes Order domi-
nated with 7 species with Genus Puntius as abundant.
From the study, it can be concluded that the reservoir
supports fairly rich fish fauna. Extension of Barul dam killed
brood fishes and juveniles, deteriorating ecological condi-
tions of spawning and rearing grounds. Local people have
to be educated on the issues of fish conservation. Finally, it
can be concluded that the Icthyofauna is very rich in the Barul
reservoir, and it can be used to increase the production of
fishes and the economic condition of local fisherman.
 Siddiqui Tasneem

Table 1: Fish fauna of Barul reservoir.

Order Family Scientific Name Vernacular Name Economic value

Cypriniformes Cyprinidae Catla catla catla FF, CO

Cirrhinus cirrosa grass carp FF, CO
Cirrhinus mrigala mrigal FF, CO
cirrhinus reba carp FF, CO
Cyprinus carpio comman carp FF, CO
Labeo rohita rohu FF, CO
Labeo fimbriatus FF, CO
Labeo calbasu karvvate FF, CO
Labeo gonius silver carp FF, CO
Barbus curmuca FF, CO
Rasborinae Rasbora daniconius dendua FF, CO
Osteoglossiformes Notopteridae Notopterus chitala chambari FF, CO
Notopterus notopterus FF, CO
Siluriformes Siluridae Callichorous papda papda FF, CO
Wallago attu goneadi FF, CF
Bagridae Mystus bleekeri FF, CO
Mystus seenghala seenghal FF, CO
Clariidae Clarias magur magur FF, CO
Perci formes Percidae Ambassis nama trash fish FF, CO
Ophiocephalidae Ophiocephalus striatus ma rail FF, CO
Ophiocephalus guchwa dhokda FF, CO
Ophiocephalus punctatus pathar chain FF, CO
Mugiloidei Mugilidae Mugil parsia FF, CO
Mugil cephalus FF,CO
Channoidei Channidae Channa guchwa dheridhok FF, CO
Mastacembelidae Channa marulius murrel FF, CO
Mastacembeloidei Mastacembelidae Mastacembelus armatus bam FF, CO

FF - Fine Food, CO - Commercial Food, CF - Coarse Food

5()(5(1&(6 Maharashtra. J. Aqua. Bio., 18(1): 47-49.

Peter, T. 1987. Fish fisheries, aquatic macrophytes and water quality in
Day, F. 1994. The Fishes of India, Burma and Ceylon. Fourth Indian Re-
inland waters. Water Quality Bull., 12(3): 103-129.
print, Vol. 1 & II, Jagmandar Book Agency, New Delhi.
Sakhare, V.B. 1999. Fisheries of Yeldari reservoir, Maharashtra. Fishing
Hora, S.L. 1956. Fish Paintings of Third Millennium B.C. from Nal
Chimes, 19(8): 45-47.
(Baluchistan) and their Zoogeographic Significance. Mem. Indian
Sakhare, V.B. and Joshi P.K. 2003. Water quality of Migni (Pangaon) res-
Mus., 14(2): 78-54.
ervoir and its significance to fisheries. Nat. Conf., Recent Trends Aquat.
Jayram, K.C. 1999. The freshwater fishes of the Indian region. Narendra
Biol., 56.
Publishing House, New Delhi, India: XVIII + 551.
Seth, A.K., Srivastava, S.K., George, N.G. and Bewtraj, J.K. 1967. Moni-
Job, J.J., David, A. and Das, K.N. 1955. Fish and fisheries of Mahanadi in
toring of certain toxic constituents in water supply. Fish Env. Him.,
relation to the Hirakund Dam. Indian J. Fish., 2(1): 1-36.
9(1): 34.
Jhingran, V.G. 1991. Fish and Fisheries of India. Hindustan Publishing
Suresh, V.R. 2003. Status of Loktak lake fisheries and approach for their
Corporation, Delhi, India.
sustainable development. Fishing Chimes, 23(3): 42-44.
Lohar, P.S. and Borse, S.K. 2003. Diversity of fish fauna in River Tapi,

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R. P. Mali and Shaikh Afsar
Department of Zoology ,Yeshwant Collage, Nanded-431 605, Maharashtra

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,1752'8&7,21 192, 216, 240 h of exposure and average value of four

Heavy metals in aquatic ecosystems often show levels above
the expected values. Wide use of heavy metals by the indus-
trial, agricultural and domestic sectors results in release of
toxic pollutants in the environment posing a serious threat
to the aquatic animals. Crabs are economically important as
they serve as an important food source to man. But, since
the last decade their natural environment is being disturbed
by the pollution. The heavy metals such as a mercury, cop-
per, zinc and cadmium are potent toxicants and cause vari-
ous physiological effects on growth, food intake, metabo-
lism and general development of aquatic animals (Kulkarni
1980, Lomte 1982, Tungare & Sawant 2000, Wankhade &
Kulkarni 2000, Rajaiah & Venkaiah 2007, Shailaja 2008,
Jagtap et al. 2009, Mali et al. 2010).
Hence, the present work was carried out to investigate
the effects of zinc sulphate on the crab Barytelphusa guerini
to evaluate the toxic effect on cardiac physiology.
0$7(5,$/6$1'0(7+2'6
Healthy and mature crabs (Barytelphusa guerini) were se-
lected for experimentation weighing 30-50 g to avoid the
effect of sex and size (Ambore 1976). The animals were ex-
posed to sublethal concentration (0.0106 ppm) of zinc sul-
phate. They were cut through the lateral side and the dorsal
carapace removed to expose the heart. They were kept in
ringer bowl filled with crab ringer and maintained for 10
minutes to allow them to recover from shock effects and for
the heart beat to get stabilized. The heart beat was seen visu-
ally and the time taken for 10 beats was determined with the
help of stop watch. The rate of heart beat was noted for
control and treated groups at 00, 24, 48, 72, 96, 144, 168,
observations was considered for calculating the rate of heart
beat.
5(68/76$1'',6&866,21
On exposure to sublethal concentration of zinc sulphate
(0.0106 ppm), the heart beat was studied at regular interval
of 24 h up to 240 h for 10 days (Table 1).
The control showed decrease up to 48 h, and later after
increasing up to 144 days, there was a gradual decrease up
to 240 h. The treated crabs showed initial increase up to 72
h, and later there was a gradual decline in the rate of heart
beat up to 240 h as plotted in graph (Fig. 1).
The literature available on the rate of heart beat in crus-
taceans indicate that the heart ratio is influenced by a number
of factors like body size, activity, light, nutritional status,
temperature, population density, oxygen and CO2 content
of the medium (Prosser & Brown 1961, Prosser 1973,
Ambore 1976, Krishnaha et al. 1987, More 1992). The im-
pact of pollutants on cardiophysiology of the freshwater crab,
Banytelphusa guerini was studied by Tonapi & Varghese
(1984). Some waterborne metals can bind to gills of fresh-
water fish and disrupt the ion-regulatory and respiratory func-
tions of the gills (Playle 1998). The physiological modula-
tions in cardiac rhythms of crab B. cunicularis, induced by
heavy metals, has been studied. Depletion in the rate of oxy-
gen consumption upon exposure to heavy metal stress might
be due to penetration of pollutants at subcellular level and
damage of gill tissue, thereby failure of an alternative com-
pensatory mechanism to achieve energy generation for com-
bating toxic stress.
In the present investigation, the initial increase in heart
 R.P. Mali and Shaikh Afsar




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Fig. 1: Effect of zinc sulphate on rate of heart beats per 10 seconds for 24-240 h.

Table 1: Rate of heart beats/10 seconds of the crab Barytelphusa guerini tain heavy metals mercury, cadmium, lead, arsenic and selenium on
after exposure to zinc sulphate for 24-240 (h). the intertidal crab, Scylla serrata. Mar. Environ. Res., 21: 109-120.
Kulkarni, K.M. 1980. The metabolic response of Barytelphusa jacquemontii
Control Crab Treated Crab to some pollutants. Geobios, 8: 133-135.
(Barytelphusa guerini) (Barytelphusa guerini) Lomte, V.S. 1982. Changes in the biochemical components of the
Exposure Weight Mean Heart Weight Mean Heart prosobranch snail, Belamia bengalensis on exposure to malathion.
(h) of crab Beats of crab Beats Proc. Symp. Physiol. Rep. Anim. Pollutants.
in (g) (H.B.) in (g) (H.B.) Mali, R.P., Afsar, S.K. and Jagtap, A.R. 2010. Impact of cadmium induced
alterations in the glycolytic potential of freshwater female crab
0 33.5 7.16 - - Barytelphusa guerini. Geobios, 37: 100-102.
24 38 5.43 36 5.05 More, A.D. 1992. Studies on some aspects of physiology of freshwater
48 42 4.67 38.5 5.93 male crab Barytelphusa guerini with special reference to inorganic
72 35 6.68 40 7.79 pollutants. Ph.D. Thesis submitted to Marathwada University,
96 47 6.91 42.5 5.82 Aurangabad.
120 44 8.29 41.5 6.45 Playle, R.C. 1998. Science Total Environment, 219(2-3): 147-163.
144 36.5 7.54 38 5.90 Prosser, C.S. and Brown, F.A. 1961. Comparative Animal Physiology. N.B.
168 38.5 6.24 34 5.52 Sounders Company, Philadelphia.
192 35.5 3.18 34.5 3.36 Prosser, C.S. 1973. Comparative Animal Physiology. N.B. Sounders Com-
216 41 3.10 39 4.12 pany, Philadelphia.
240 45.5 2.80 47.5 3.37 Rajaiah, V. and Venkaiah, Y. 2007. Effect of parathion on esterase patterns
of Channa punctatus. J. Aqu. Biol., 22(1): 181-185.
Ketpadung, R. and Olan, N.T. 2006. Changes in oxygen consumption and
rate in treated crabs may be due to accelerative metabolic heart rate of the blue swimming crab, Portunus pelagicus (Linnaeus,
activity of the animal and decline in the rate of heart beat 1766) following exposure to sublethal concentrations of copper. Jour-
indicating that animals try to settle down to the toxic nal of Environmental Biology, 27(1) 7-12.
medium. Shailaja, V., Madhuri, E., Ramesh Babu, K., Rama Krishna, S. and Bhasker,
M. 2008. Study of protein metabolism in hepatopancreas and muscle
$&.12:/('*(0(17 of prawn Penaeus monodon on exposure to altered pH media. J.E.F.
1: 85-92.
The author is thankful to Principal, Yeshwant College, Tonapi, G.T. and Varghese, G. 1984. Cardiophysiological responses of the
Nanded, Maharashtra for providing facilities. crab, B. cunicularis (Westwood) to three common pollutants. Ind. J.
Expt. Biol., 22: 548-549.
5()(5(1&(6 Tungare, S.M. and Sawant, A.D. 2000. Physiological effects of heavy met-
als on prawns. International Conference on Probing in Biological Sys-
Ambore, N.E. 1976. Studies on some aspect of physiology in freshwater tems, Mumbai, Feb. 2000, pp. 139.
crab, Barytephusa guerini with special reference to sex and size. Ph.D. Varghese, G., Naik, P.S. and Katdare, M. 1990. Respiratory responses and
Thesis submitted to Marathwada University, Aurangabad. blood sugar level of the crab, Barytelphusa guerini expressed to mer-
Jagtap, A.R., Afsar, S.K., Kothole, S.D. and Mali, R.P. 2009. Effect of pol- cury, copper and zinc. Ind. J. Exp. Biol., 30(4): 308-312.
lutants from car washing centre on oxygen consumption in freshwater Wankhade, V.M. and Kulkarni, K.M. 2000. Liver protein profiles of the
fish, Channa punctatus. J. Aqu. Biol., 24(2): 189-192. fish Channa orientalis exposed to endosulphan. J. Aqua. Biol., 15(1
Krishnahan, A.P., Rage, M.S. and Joshi, A.G. 1987. Toxic effects of cer- & 2): 89-91.

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Ayan Chandra, Gaurav Saha*, Sayantan Goswami*, Anwesha Saha*, Soumi Sinha Roy*, Payel Ganguly*, Avishek
Ghosh*, Kaveri Banerjee*, Radhika Saraf, Arpita Banerjee** and Arup Kr. Mitra*
Department of Statistics, St. Xavier’s College (Autonomous), Kolkata-700 016, W. B., India
*Department of Microbiology, St. Xavier’s College (Autonomous), Kolkata-700 016, W. B., India
**Department of Respiratory Medicine, Nil Ratan Sircar Medical College, Kolkata, W.B., India

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,1752'8&7,21 But in Kolkata and other metropolitan cities, these stand-

The surge of urbanization has engulfed Kolkata to the full-
est. Wherever we put our eyes, we find new high rise build-
ings being built, more and more vehicles hitting the streets,
and more and more industries being set up. There are both
advantages and disadvantages of the increased urbanization.
While the benefits are manifold, there is one disadvantage
in the form of increasing air pollution. Even if we do not
consider other kinds of pollution, the effect of air pollution
in itself is the maximum, both in intensity and in destruc-
tion. Air pollution is generally measured by the parameters,
suspended particulate matter (SPM), respirable particulate
matter (RPM ), oxides of sulphur (SOx) and oxides of nitro-
gen (NOx).
SPM plays a key role in determination of the amount of
air pollution in metropolitan cities. The standards have been
established by the CPCB (Central Pollution Control Board).
ards are often violated, hence, the government runs scans of
the concentration of SPM regularly to keep its increasing
values in check. CPCB has set up standards of 120 µg/m3
and 60 µg/m3 for RPM in the industrial and residential areas
respectively. The government also tries to undertake vari-
ous methods in order to keep these standards in check.
The Kolkata has a large number of vehicles. As the old
vehicles are not well maintained, they cause high pollution
as compared to the newer vehicles. This invoked the gov-
ernment to impose a ban on such vehicles, which are more
than 15 years old. This ban came into effect on August 1,
2009, and 22000 trucks, 6000 taxis, 32000 auto rickshaws,
500 mini buses and 2500 private buses were banned from
the streets of Kolkata. After the ban was imposed, the air
improved from the earlier pollution levels. So, we under-
took an attempt to prove whether the ban was really a suc-
cess or not and whether it had any relation to the diseases
 Ayan Chandra et al.

Table 1: Year-wise comparison of various air pollutants (µg/m3) for the years 2008, 2009 and 2010.

Winter(January-February) Summer(April-May) Monsoon(June-July)

2008 2009 2010 2008 2009 2010 2008 2009 2010

RPM 145.3 171.1 184.4 51 47 43.7 34 34.9 33.5

SPM 302.4 334.3 383.1 132.3 116.3 114.4 96.8 93 92
NOx 84.9 85.5 104.1 57.6 45.3 48 44.3 43.9 42
SOx 7.8 10.1 12.2 5.2 5.5 7.4 4.9 5.2 5.1

Table 2: Patients affected due to air pollutionas as per data collected from Data collection for disease incidence during pre-banning,
the three hospitals.
banning and post-banning time periods: All the data were
Year Seasons number of Sample size of collected following the interview method in which data were
patients each year collected from 3 hospitals across the city of Kolkata by di-
rect interviews with the hospital authorities. From this we
2008 Winter 142 366
Summer 126
came to know about the patients suffering from diseases due
Monsoon 98 to air pollution. The doctors distinguished between the vari-
2009 Winter 153 365 ous kinds of diseases by Pulmonary Function Test (PFT).
Summer 117 Firstly, data related to patients were obtained for their name,
Monsoon 95
2010 Winter 157 377 age, gender, height, weight and smoking history. With the
Summer 119 help of spirometry (Hancox & Whyte 2005), the following
Monsoon 101 parameters were obtained: FVC (Forced Vital Capacity),
Total Sample Size 1108 FEV1 (Forced Expiratory Volume on 1st sec) and FEV1/FVC.
Considering these values, finally diagnosis was made as the
caused by air pollution. A survey was performed in various
normal, the obstructive airways or the restrictive lung disor-
renowned hospitals of Kolkata. We have collected data in
der. The differentiation of COPD from asthma was done by
three different seasons, summer (April-may), monsoon (June-
considering the Reversibility Criteria as mentioned by the
July) and winter (January-February) in the years 2008, 2009
American Thoracic Society.
and 2010. t-test, ANOVA and box plots tests were performed
to determine correlation between the diseases and the ban,  .


 
and to find out whether the ban on old vehicles has been a
success or failure. Year-wise comparison of RPM, SPM, NOx, SOx: Year-
wise comparison of the four parameters was performed in
-

.


-/ winter, summer and monsoon seasons. The maximum value
of all the parameters was found in the months of January-
Air analysis: Air samplers having glass impingers contain-
February of 2010, and the minimum values in June-July of
ing specific solvents for measuring the levels of NOx and
the same year (Table 1).
SO2 in ambient air were used. In order to capture the
particulate matter present in the ambient air, a filter paper Season-wise comparison of RPM, SPM, NOx, SOx: Sea-
was placed in front of the absorbing jet. son-wise comparison of the four parameters was performed
for the three years separately. This showed that the winter
Sulphur dioxide was determined by modified West and
season has the maximum values for all the three years, with
Geake method. Nitrogen oxides were determined by absorb-
2010 showing the highest values, and monsoon season the
ing them in sodium hydroxide-sodium arsenite solution and
minimum values for all the three years with 2010 showing
later analysing by colourimetric method. SPM and RPM (par-
the least concentrations (Figs. 1, 2, 3).
ticles lesser than 10 µm) were determined by filter paper dif-
ference method using specific filter papers after passing Patients affected by air pollution as per data collected
known volume of air through them. from the three hospitals: The analysis of the three hospi-
tals showed that the highest number of patients affected by
Statistical tests: The tests like t-test , ANOVA and box plates
air pollution were in the winter seasons of all the three years
were carried out for correlating the disease incidence with
with maximum in 2010. The sample size year-wise was 366
levels of pollutants by standard statistical methods. The dif-
for 2008, 365 for 2009 and 377 for 2010, hence the total
ferences in the level of the air pollutants during the three
sample size was 1108 for the three hospitals (Table 2,
seasons were also tested by ANOVA to check whether they
Figs. 4, 5).
are significant or not.

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Table 3: Patients suffering from respiratory disease, lung cancer and cardiac disease in the three seasons of 2008, 2009 and 2010.

Winter Summer Monsoon

Diseases 2008 2009 2010 2008 2009 2010 2008 2009 2010

Respiratory Disease 108 114 117 95 88 86 71 68 71

Lung Cancer 9 10 9 9 10 8 8 9 8
Cardiac Disease 25 29 31 22 19 25 19 18 22

Table 4: Patients suffering from asthma, restrictive lung disorder and COPD in the three seasons of 2008, 2009 and 2010.

Seasons Winter Summer Monsoon

YearsDiseases 2008 2009 2010 2008 2009 2010 2008 2009 2010

Asthma 38 41 42 34 30 29 27 26 28
Restrictive Lung Disorder 37 38 39 31 29 27 24 23 21
COPD 33 35 36 30 29 30 20 19 22

  

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Patients suffering from respiratory diseases in the three
seasons: There are three diseases selected for analysis of the
effect of air pollution on health of the citizens of Kolkata.
The analysis shows that the patients suffering from respira-
tory disease are highest. The number of cardiovascular dis-
ease patients is the second highest and the number of lung
cancer patients is the least. The maximum number of pa-
tients was found in the year 2010 which is after the ban of
old vehicles (Table 3).
Patients suffering from respiratory disorders in the three
seasons: The three main respiratory disorders are asthma,
restrictive lung disorder and chronic obstructive pulmonary

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disorder (COPD). The maximum number of patients is for
asthma during the winter season of 2010. In fact, all the dis-
eases affected the citizens to the greatest in the winter of
2010, a few months after the ban on old vehicles was im-
posed, showing that the ban has failed to mitigate the air
pollution effects (Table 4).
Analysis of variance (ANOVA) of the four parameters:
The ANOVA of the four parameters has been performed to
see whether their levels differ significantly or not from the
average levels in the three seasons. It was found that all the
four parameters differ significantly from their average
values for all the three seasons of the three years.

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Box plots of the SPM, RPM, SOx and NOx: Four box plots
were obtained by calculating the p-values for all the four
parameters. It was found that the average levels of SPM and
RPM differ significantly from their standard values but SOx
and NOx do not differ much from the standard values (Figs.
6, 7, 8, 9).
Kolkata is among the most polluted cities of the world.
The major reason for the highly increased air pollution load
in the city during the 2000-2008 period was huge increase
in the number of private taxis, buses, autos, cars, etc. The
city also has an additional problem of having very old vehi-
cles.
The Supreme Court banned all the vehicles greater than
15-year old from plying on roads of Kolkata, which was made
effective from August 1, 2009. About 32000 auto rickshaws,
2500 private buses, 6000 taxis, 500 mini buses and 22,000
trucks were banned. However, there was no guarantee that
the vehicles less than 15-year old are not going to increase
air pollution.
In order to investigate the air pollution load in Kolkata,
an year-wise comparison of the four air pollution param-
eters SPM, RPM, NOx and SOx was undertaken consider-
ing three periods of before the ban, during the ban and after
the ban. The season-wise comparison showed that the win-
ter season of 2010 had the highest value of particulate mat-
ter, while monsoon of 2010 had the lowest values for the
same. The season-wise comparison showed that winter sea-
son for all the three years has highest concentration of all
the four parameters, while the monsoon season the lowest
values in all the three years.
A few questionnaires were also used for survey in the
three hospitals to find whether air pollution has caused any
disease among the citizens. The doctors gave a list of pa-
tients who were suffering from diseases caused due to air
+

(
Fig. 9: Box plate of SOx.

pollution. The number of patients was typically found to be
the highest in the winter season of 2010 with a total of 157
patients. This is really striking as the highest number of pa-
tients seems to have appeared when the ban on old vehicles
has been imposed in Kolkata. This really makes the ban an
unsuccessful one. The other two seasons, summer and
monsoon, also show that the ban has not been that much
successful as the number of patients after decreasing for the
year 2009 after the ban for some time, increased again in the
next year 2010.
The air pollution is a major cause for various kinds of
diseases particularly the respiratory diseases like asthma,
restrictive lung disorder and chronic obstructive pulmonary
disorder, lung cancer, and cardiac diseases (Ghose et al.
2005). These diseases have taken a toll on the citizens maxi-
mum during the winters (2010) and the minimum during
the monsoons (2009). This is probably due to the high con-
centration of the particulate matter present in the atmosphere
during winter, and lowest in monsoon. The effect of the ban
could not be understood properly on these patients as the
study has to be on a longer term basis. The air pollution can
also cause mutations, which are time consuming hence, the
effect of air pollution was seen during the time of 2008 but
could not be analysed properly after the ban as the time span
was short.
The pulmonary diseases are common outcome of the high
levels of air pollution (Seaton 1995). The particulate matter
concentration is generally high in the atmosphere and if par-
ticles are of the respirable size (< 0.65µ), then they get re-
spired into the lungs and get deposited inside the same. Thus,
hindering the proper activity of the lungs like gas exchange
in the alveoli or the release of surfactant type of chemicals
by bronchioles, which lead to various kinds of respiratory
disorders. In asthma, the spastic contraction of the
bronchioles occurs during expiration. This occurs due to the
,

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x9RO1R
 Ayan Chandra et al.
hypersensitivity reactions caused by air pollutants that act
as allergens and the particulate matters get deposited on the
bronchial walls leading to the improper release of the
surfactants (DPPC-dipalmitoyl phosphatidyl choline)
thereby leading to improper contraction and relaxation of
the muscles.
In the restrictive lung disorder, the particulate matters
get deposited in the alveoli leading to improper gas exchange.
The lung tissues do not get proper oxygen supply and the
carbon dioxide content increases which leads to improper
cell functioning and finally lung failure.
COPD is mainly comprises of two diseases, chronic
bronchitis and emphysema. The particulate matter and other
primary air pollutants cause the air passage to get constricted
and the destruction of the alveoli and terminal bronchioles.
These two diseases are the result of inflammation and tissue
degradation (Russell Michael 2006). The NO2 is a potent
agent for PAN (peroxy acetyl nitrate) formation. It is a pow-
erful respiratory irritant, lachrymator and mutagenic in na-
ture (Lui et al. 1999). The peroxy acetyl nitrate, if inhaled,
leads to destruction of the tissues of lungs leading to inflam-
mation and the respiratory distress.
The particulate matters, SOx and NOx cause an enhanced
coagulation or thrombosis. They propel arrhythmia and acute
arterial vasoconstriction due to deposition of particulate
matter in the blood vessels leading to atherosclerosis. This
leads to improper functioning of heart and its related vascu-
lar system (Brook et al. 2004). This leads to cardiovascular
diseases. This may lead to a fatal state of myocardial infarc-
tion. The SOx and NOx are also the main causative agents
for the lung cancer apart from few particulate matters like
lead, cadmium, etc.
Our main goal was to determine the effectivity of the old
vehicular ban on the streets of Kolkata. It was found that
after the ban there was a dip in the levels of SPM, RPM,
SOx and NOx. But with time, as more and more vehicles
entered the roads of Kolkata and the people became forget-
ful of the ban, the levels of these four air pollutants increased
and are now showing a rise. The increasing pollution can be
controlled if the ban is again imposed strictly. The West Ben-
gal Pollution Control Board conducts air pollution surveys
regularly and launched the ICEF (India Canada Environment
Facility) project to reduce SPM and RPM levels (Chakrabarty
& Bhattacharya 2004). Further, decentralization of office ar-
eas, improvement of road conditions, regulation of traffic
and proper implementation of the ban on polluting vehicles
9RO1Rx
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is mandatory to improve air pollution levels in the city. The
greater the government promotes the usage of eco-friendly
fuel, the better it will be for the air of Kolkata. The govern-
ment has to strengthen its Inspection and Maintenance (I&M)
of public and private transport. The WBPCB and CPCB
(Central Pollution Control Board) are working hard to im-
prove the air quality, but ultimately, it is duty of citizens to
abide by the rules if we have to breathe a cleaner air.

.0-
The authors express their greatest gratitude to the Principal
Rev. Fr. Dr. Felix Raj and the Vice-Principal, Rev. Fr. Jimmy
Keepuram of St. Xavier’s College, Kolkata for support. The
help and guidance rendered by the Dr. Surabhi Dasgupta
HOD, Department of Statistics is also gratefully acknowl-
edged. We would also like to thank the WBPCB and Prof.
Sumitra Basu Thakur for providing various information that
has helped us in this project.
1
Brook, D. Robert, Franklin Barry, Chair, Cascio Wayne, Hong Yuling,
Howard George, Lipsett Michael, Luepker Russell, Mittleman Murray,
Samet Jonathan, Smith C. Sidney, Jr., Tager Ira 2004. Air Pollution
and Cardiovascular Disease, A Statement for Healthcare Profession-
als from the Expert Panel on Population and Prevention Science of the
American Heart Association. Cir., 109: 2655-2671.
Chakrabarty Arijita, Bhaduri Satarupa, Ganguly Shibayan, Mukherjee
Gairik, Das Barenya and Chandra Ayan 2010. Alleviation of stress
induced hyperglycemia in urban population. Abstract published in 97th
Indian Science Congress, Kerala.
Chakrabarty, D. and Bhattacharya, P. 2004. Air quality management strat-
egy for Kolkata city. Indian Journal of Air Pollution Control, VII(1):
49-56.
Chaulya, K. S. 2004. Spatial and temporal variations of SPM, RPM, SO2
and NOx concentrations in an opencast coal mining area. J. Env.
Management, 70(1): 1-14.
Ghose, K. Mrinal, Paul R. and Banerjee, K. R. 2005. Assessment of the
status of urban air pollution and its impact on human health in the city
of Kolkata. Environment Monitoring and Assessment, 108: 151-167.
Hancox Bob, Whyte Ken 2005. Pocket Guide to Lung Function Test.
2nd. Edition, McGraw Hill, pp. 195.
Liu, G.Y., Chen, K. J., Lin-Shiau, S. Y. and Lin, J.K. 1999. Peroxy acetyl
nitrate-induced apoptosis through generation of reactive oxygen spe-
cies in HL-60 cells. Molecular Carcinoq., 25(3):196-206.
Mukherjee, Gairik, Das Barenya, Nandi Surajit, Saha Anwesha, Roy Sinha
Soumi, Islam Rubina and Mitra Arup Kumar 2010. Impact of banning
of old vehicles from the streets of Kolkata. Abstract in 97th Indian
Science Congress, Kerala.
Russell Michael 2006. An article on COPD-definition, symptoms, diagno-
sis and treatment. Asbestos Times, 21st June, 2006.
Seaton, P. 1995. Particulate air pollution and acute health effects. The Lan-
cet, 345(8943): 176-178.
,

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