Survey and Evaluation of Oleaferous Freshwater Algae of Dal Lake Ecosystem

350 Pages·2014·16.53 MB·English

by  Javeed Ahmad Lone

Survey and Evaluation of Oleaferous Freshwater Algae of
Dal Lake Ecosystem

Javeed Ahmad Lone
(2010-332-D)

Division of Environmental Sciences
Faculty of Postgraduate Studies
Sher-e-Kashmir University of Agricultural Sciences &
Technology of Kashmir

2014
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Survey and Evaluation of Oleaferous Freshwater Algae of
Dal Lake Ecosystem

Javeed Ahmad Lone
(2010-332-D)

Thesis

Submitted to

The Faculty of Post-graduate Studies
Sher-e-Kashmir University of Agricultural Sciences and
Technology of Kashmir
in partial fulfilment of requirement for the award of the degree of

Doctor of Philosophy in Environmental Sciences

2014
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Whose blessi ngs and constant
encou ragement
have been always w ith me throughout my

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Sher-e-Kashmir
University of Agricultural Sciences and Technology of Kashmir
Division of Environmental Sciences, Shalimar Campus, Srinagar -
190 025

Certificate – I

This is to certify that the thesis entitled “Survey and
Evaluation of Oleaferous Freshwater Algae of Dal Lake
Ecosystem” submitted in partial fulfilment of the requirements for the
award of the degree of Doctor of Philosophy in Environmental
Sciences, to the Faculty of Postgraduate Studies, Sher-e-Kashmir
University of Agricultural Sciences and Technology of Kashmir, is a
record of bonafide research work carried out by Mr. Javeed Ahmad
Lone (Regd. No. 2010-332-D) under my supervision and guidance.
No part of the thesis has been submitted for any other degree or
diploma.
It is further certified that any help or information received
during the course of investigation have duly been acknowledged.

Dr. F.A. Lone
Chairman
Advisory Committee

Endorsed

Prof. & Head,
Division of Environmental Sciences

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Sher-e-Kashmir
University of Agricultural Sciences and Technology of Kashmir
Division of Environmental Sciences, Shalimar Campus, Srinagar-
190 025

Certificate – II

We, the members of the Advisory committee of Mr. Javeed
Ahmad Lone (Regd. No. 2010-332-D), a candidate for the degree of
Doctor of Philosophy in Environmental Sciences, have gone
through the manuscript of the thesis entitled, “Survey and
Evaluation of Oleaferous Freshwater Algae of Dal Lake
Ecosystem” and recommend that it may be submitted by the student
in partial fulfilment of the requirements for the award of degree.

Advisory Committee

Chairman Dr. F.A. Lone
Associate Professor,
Division of Environmental Sciences
Members
Dr. Javeed Iqbal Ahmad Bhat
Associate Professor,
Division of Environmental Sciences

Dr. G.H. Dar
Professor, Division of Plant Pathology

Dr. J.A. Wani
Associate Professor, Division of Soil Science

Dr. Showkat Maqbool
Assistant Professor, Division of Agri-Statistics

Dean PG Nominee Dr. Tariq Raja
Associate Professor, Division of Agri-Statistics

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Sher-e-Kashmir
University of Agricultural Sciences and Technology of Kashmir
Shalimar Campus, Srinagar – 190 025

Certificate – III

This is to certify that the thesis, “Survey and Evaluation of
Oleaferous Freshwater Algae of Dal Lake Ecosystem” submitted
by Mr. Javeed Ahmad Lone (Regd. No. 2010-332-D) to the Faculty
of Postgraduate Studies, Sher-e-Kashmir University of Agricultural
Sciences and Technology of Kashmir, in partial fulfilment of the
requirements for the award of the degree of Doctor of Philosophy in
Environmental Sciences was examined and approved by his
Advisory Committee and external examiner on ………………..

Chairman External Examiner
Advisory Committee

Prof. & Head
Division of Environmental Sciences

Director Education,
SKUAST-Kashmir, Shalimar, Srinagar

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Sher-e-Kashmir
University of Agricultural Sciences and Technology of Kashmir
Division of Environmental Sciences, Shalimar Campus, Srinagar-
190 025

Name of the student : Javeed Ahmad Lone
Registration No. : 2010-332-D
Major subject : Environmental Sciences
Minor subjects : Soil Science/Forestry
Major advisor : Dr. F.A. Lone,
Associate Professor,
Division of Environmental Sciences,
SKUAST-Kashmir, Shalimar
Title of the Thesis : “Survey and Evaluation of Oleaferous
Freshwater Algae of Dal Lake Ecosystem”

ABSTRACT
In the present study investigations on the various oleaferous algae of Dal
Lake were carried out to identify the promising microalgal isolates for biodiesel
production which is very much relevant to the need of biofuel industry. During the
study 96 water samples and 96 algal samples were collected during the four
seasons of the year from six different sites (DLS-I = Nehru Park, DLS-II = Bren
Laam, DLS-III = Charchinar, DLS-IV = Hazratbal, DLS-V = Nagin, DLS-VI =
Ranawari) of the lake. The water chemistry revealed that the lake is undergoing
tremendous cultural eutrophication because of different anthropogenic activities
viz., waste water from house boats and local population, agricultural runoff from
the floating gardens, catchment areas and tourist activities. DLS-VI was the most
polluted site of the lake during the four seasons of the year. However, the
biological studies of the lentic ecosystem revealed that the lake is also rich in
microalgal flora. A total of 91 algal genera were identified comprising of 217
species, 41 varieties and 8 forma. Maximum number of species belongs to
Chlorophyceae (149) with 30 varieties and 8 forma. The Chlorophycean algae of
Dal Lake was best represented in summer and autumn months and lowest in the
winter followed by spring season. In the class Bacillariophyceae, 30 species and 5
varieties were recorded showing their peaks of standing crop during winter
months. Cyanophyceae was the third largest class with 28 species and 2 varieties
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and showed their peak abundance during summer and autumn seasons with the
maximum standing crop at DLS-VI. Among the Euglenophyceae, 4 genera, 9
species with 4 varieties were recorded and all these species were found in
appreciable numbers at DLS-VI during the autumn season. Rhodophyceae was
monotypic as it was represented by only one taxon and is a new record to the
phycological studies of India. To identify the most promising biodiesel microalgal
strains for our future research programme thirteen microalgal isolates were also
successfully isolated and purified from the six different sites of the Dal Lake and
were subjected to lipid analysis using slight modified Folch method (1957). The
peak lipid content in the isolates ranged from 4.63-30.99 per cent. Amongst all,
two microalgae Scenedesmus dimorphus and Scenedesmus quadricauda were
selected for further studies as these two isolates possess appreciable amount of
lipids 30.99 per cent and 28.61 per cent respectively. The growth analysis pattern
of these two robust algae in the BBM media showed that both the species are fast
growing and reached a stationary phase on 14th day of incubation and are suitable
for high-density culture. The photosynthetic pigments were calculated using two
set of equations viz., Arnon’s formulas and Wellburn equations. High amount of
total pigments (mg g-1 fw) were calculated when Wellburn equations were applied
(S. dimorphus = 19.806 and S. quadricauda = 27.099) and Arnon’s equations (S.
dimorphus = 7.274, S. quadricauda = 9.713) were found to be inefficient when
using DMSO as a solvent. The study also revealed that maximum protein content
was found in S. quadricauda (13.026%) followed by S. dimorphus (8.284%).
Based upon the fast growth and maximum lipid content, the two promising
species of microalgae were selected for large scale biomass production in self
made 25 liter lab scale photobioreactors having 15 liters BBM media. Both the
species thrived very efficiently and biomass was harvested after a period of five
weeks of incubation using different harvesting techniques like flocculation,
centrifugation, lypholisation and oven dry. The lypholized biomass was subjected
to lipid extraction by Soxhlet (1875) and Folch method (1957) using same solvent
chloroform: methanol (2:1). In Folch method 30.99 per cent oil content was
reported from S. dimorphus while as 28.61 per cent was reported from S.
quadricauda. On the other hand in Soxhlet extraction 27.29 per cent oil was
reported in S. dimorphus while as 24.75 per cent oil was reported from S.
quadricauda. The results reveal that in case of green microalgae Folch method is
more efficient as compared to Soxhlet in terms of oil extraction as in the former
there is complete mechanical disruption of cells. The microalgal oil of both the
species was subjected to fatty acid (FA) analysis and the fatty acid methyl ester
(FAME) profiles showed that both the species possess appreciable amounts of
major FA with carbon chain length of C16 to C18 viz oleic acid 21.1 per cent,
26.2 per cent, palmitic acid 18.9 per cent, 17.8 per cent, and linoleic 13.1 per cent,
13.8 per cent making them suitable feedstock for the production of good quality
biodiesel. The total amount of fatty acid methyl ester of the S. dimorphus was
86.2 per cent and that of S. quadricauda was 85.7 per cent with 13.8 per cent and
14.3 per cent hydrocarbons and unidentified. The quality parameters of both the
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microalgal oil like degree of unsaturation (DU), cetane number (CN), iodine value
(IV), saponification value (SV) were within the limits of ASTM D6751, EN
14214 and ANP 255 standards respectively. The physico-chemical characteristics
of oil obtained from these two tested microalgae viz., colour, density and viscosity
were too high and did not pass the standards. The highly dense and viscous oils of
both the microalgae reveal that transesterification is an important step to minimize
these physico-chemical characteristics of the oil and conversion of the algal oil
into biodiesel. Over all our results suggest that both the species of Scenedesmus
are the promising isolates for producing high quality biodiesel.

Key words: Biodiesel, Dal Lake, Fatty acids, Lipids, Microalgae, Scenedesmus
dimorphus, Scenedesmus quadricauda

Signature of Student Signature of Major Advisor

Dated:____________ Dated:_________________

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ACKNOWLEDGEMENT

I bow in reverence to the beautiful creator of the universe who is almighty Allah (Subhana–Wa–
Tallah) for inspiration and for every success whose being benediction gave me all required zeal for
the completion of this work.
V ocabulary in spite of all developments is still imperfect to express the
feelings of gratitude, because when expressed in a word is only half its
acknowledgement. Perhaps certain feelings of best expressed when felt unsaid.
Yet I made an effort to express my overwhelming sense of gratitude for that has
made this task possible.
It is a matter of immense pleasure to acknowledge my debt to my revered
Chairman Dr. F. A. Lone, Associate Professor, Division of Environmental
Sciences, SKUAST-Kashmir, Shalimar. It is because of his priceless intellectual
innovative guidance, valuable suggestions, constructive criticism and
constructive ideas for having given me complete independence, affectionate
encouragement to put my desire and thought, which paved the way for the
successful completion of this work. It is my fortune to get an opportunity to
work under the supervision of such great personality. I also wish to extend my
sincere thanks to my Chairman for suggesting me to work at Algology
Laboratory, Plant Biodiversity, Systematics and Herbarium Division of
National Botanical Research Institute, Lucknow.
I deem it to be my proud privilege to express my deep sense of gratitude
and indebtedness to modest, generous and courteous personality Dr. M. R.
Suseela, Scientist-F and Head, Algology Laboratory, National Botanical
Research Institute, Lucknow for her attentive guidance, noble and humble
advise, constructive suggestions, keen and sustained interest during the entire
period of investigation. She has been a perennial source of inspiration and
encouragement from the beginning till the completion of the thesis, without
which this work could not have seen the light of the day.
I have warmest regard and am highly thankful to the members of my
advisory committee, Dr. Javeed Iqbal Ahmad Bhat, Associate Professor,
Division of Environmental Sciences; Dr. G. H. Dar, Professor, Division of
Plant Pathology; Dr. J. A. Wani, Associate Professor, Division of Soil Science;
Dr. Showkat Maqbool, Assistant Professor, Division of Agri-Statistics; Dr.
Tariq Raja, Associate Professor, Division of Agri-Statistics (Dean P.G.
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