Diversity among the yeasts isolated from naturally fermented cashew apple juice

208 Pages·2016·3.21 MB·English

“Diversity among the yeasts isolated from naturally
fermented cashew apple juice”
Submitted to
GOA UNIVERSITY
For the award of the degree of
DOCTOR OF PHILOSOPHY IN MICROBIOLOGY
by
Supriya Nagesh Prabhu Khorjuvenkar
Work carried out under the guidance of
Dr. S. B. Barbuddhe
(Guide)
Principal Scientist (Veterinary Public Health)
National Institute of Biotic Stress Management
Raipur, Chhattisgarh, India
July 2016
Dedicated to
My loving Parents
Mr. Nagesh Prabhu Khorjuvenkar
Mrs. Geeta N. Prabhu Khorjuvenkar
My sister
Diksha N. Prabhu Khorjuvenkar
Certificate
Certified that the research work embodied in this thesis entitled
“Diversity among the yeasts isolated from naturally fermented cashew
apple juice” submitted by Ms. Supriya Nagesh Prabhu Khorjuvenkar for
the award of Doctor of Philosophy degree in Microbiology at Goa
University, Goa, is the original work carried out by the candidate herself
under my supervision and guidance.

Research Guide
Dr. S. B. Barbuddhe
M.V.Sc., Ph.D.
Principal Scientist
National Institute of Biotic Stress Management, Raipur,
Chhattisgarh, India
Statement
I hereby state that this thesis for Ph.D. degree on “Diversity among the
yeasts isolated from naturally fermented cashew apple juice” is my original
contribution and that the thesis and any part of it has not been previously
submitted for the award of any degree/diploma of any University or Institute. To
the best of my knowledge, the present study is the first comprehensive work of its
kind from this area.

Supriya Nagesh Prabhu Khorjuvenkar
Ph.D. Student
Department of Microbiology
Goa University,
Goa
Acknowledgment
I am extremely happy and pleased that my dream of entering into Research work
is coming true. I now know why great Scientist forget their social life once they enter
into science laboratory. But my work in the laboratory and outside was made easy by Dr.
S.B. Barbuddhe, Principal Scientist at ICAR-National Institute of Biotic Stress
Management, Raipur.
He has a neck of extracting and exploring the hidden skills and talents in his
students. He is very kind but at the same time, stern and disciplined as far as perfection
in the work is concerned. A person who can be a model to his students. I am grateful to
you Sir for all efforts you have invested in me.
I am thankful to my VC’s Nominee, Dr. Sandeep Garg, Associate Professor,
Department Of Microbiology, Goa University, for his constructive suggestions and
taking every step to go deep and deep into the matter related to topic and research.
My gratitude goes to Dr. Saroj Bhosle, Retired Dean of Life Science and
Professor of Microbiology Department, Goa University for encouraging and supporting
me to take up research work
I thank Dr. N.P. Singh, Director, Indian Council of Agricultural Research
Complex for Goa for providing facilities to carry out research work at the Institute. I am
thankful to Dr. Z.B. Dubal, Senior Scientist, IVRI, Izatnagar, for his co-operation during
the work.
I also thank Mr. Salim Veljee, Director, Directorate of Food and Drug
Administration, for allowing me to use the chemicals and conduct a part of my research
work in the FDA laboratory. I would like to express my gratitude to Mrs. Nilima Mishal,
Scientific Officer, FDA Laboratory, for her assistance and suggestions whenever
necessary.
I am also thankful to Dr. Swapnil Doijad, who inspite of his busy schedule gave
his valuable time for me and provided an insight into the writing of thesis. I am also
thankful to my colleagues Abhay Raorane, Ajay Pathak, Dr. Shubhada Chothe,
Prathamesh Ghavnalkar and Priyanka Naik Parrikar for their support and useful
suggestions and advices during the research work.
The collection of fermented cashew apple juice samples from fields was possible
because of the help and co-operation of various persons especially, Prof. Deepak
Karmalkar, Mr. Narayan Mangeshkar, Mr. Roy, Mr. Andrew, Mr. Bhakta, Mr. Kamat,
Mr. Hanumant Pednekar and Mr. Victor.
I must thank Dr. Swapnil Arsekar M.D. (Medicine) and Dr. Sarika Arsekar M.D.
(Gynaec.) for recommending ways and means to relive stress and strain to realize the
dream. Prof. Rajendra Arsekar (Ex. Principal of D.M’s College), my uncle, for
reminding me every now and then how important it is to submit the thesis well in time.
Whenever I was depressed, Ms. Sujata Arsekar, my aunt gave examples of others to
make me lively. Thank you my uncle, Mr. Mahableshwar Prabhu Khorjuvenkar for
trying to build up my courage to complete research work and Mr. Ashok Prabhu
Khorjuvenkar for his great support.
Friends make life worthwhile. I am grateful to Priya Narvekar, Surbhi
Mandrekar, Nandini Dabholkar, Krunal Dangui, Dr. Teja Gaonkar, Dr. Trelita, Dr.
Meghanath Prabhu, Adarsh Shettigar and my cousins Richa, Samiksha, Raj, Vipul,
Goldie, Rahul and Gayatri for their moral support. I am indeed indebted to my sister
Diksha for her invaluable help and sharing views to boost my pensive mood and helping
me to collect cashew apple juice samples. Shriyan and Sandesh, Diksha’s colleagues,
were a great help to me. Thank you.
The whole credit of joining research field goes to my father, Mr. Nagesh and my
mother Mrs. Geeta. I thank for their understanding, wisdom, patience, enthusiasm and
encouragement and for pushing me further than I thought I could go. I would like to
place on record that their trust and belief in me will be rewarded.
I must express my gratitude to my late grandparents and my late aunt Pramodini
for nurturing me and showing little extra affection so that I may attain my goal.
I am thankful to the Almighty for giving me good health for all these years and
helping me to complete my research work and keeping me grounded throughout the
process.

Supriya Nagesh Prabhu Khorjuvenkar
Abbreviations

4-MUG 4-methylumbelliferyl-b-d-glucoside
BHI Brain Heart Infusion
BIS Bureau of Indian Standards
BLAST Basic Local Alignment Search Engine
CaCl Calcium chloride
2
CAJ Cashew apple juice
CellA Cellulase Activity
CMC Carboxy Methyl Cellulose
CO Carbon dioxide
2
Conc. H SO Concentrated Sulphuric acid
2 4
CTAB Cetyl Trimethyl Ammonium Bromide
DNA Deoxyribo Nucleic Acid
EEA Extracellular Enzyme Activity
FDA Food and Drug Administration
FeSO .7H O Ferrous sulphate
4 2
FSSAI Food Safety and Standards Authority of India
G.I Geographical Indication
GlucA Glucosidase Activity
H PO Phosphoric acid
3 4
HCl Hydrochloric acid
ICAR Indian Council of Agricultural Research
IS Indian Standards
ITS Internal Transcribed Spacer
KMnO Potassium permanganate
4
LipA Lipase Activity
MgSO .7H O Magnesium sulphate
4 2
MTCC Microbial Type Culture Collection
mtDNA Mitochondrial DNA
Na S O Sodium thiosulphate
2 2 3
NaCl Sodium chloride
NCBI National Center for Biotechnology Information
PCR Polymerase Chain Reaction
PDA/PDB Potato Dextrose Agar/Potato Dextrose Broth
PectA Pectinase Activity
PrA Protease activity
RFLP Restriction Fragment Length Polymorphism
RNA Ribo Nucleic Acid
SDA Starch Degrading Activity
TA Titratable Acidity
UrA Urease Activity
VA Volatile Acidity
YEPG Yeast Extract Peptone Glucose
YPD Yeast extract Peptone Dextrose
Units of Measurement

cfu colony forming unit
g Gram
g/L gram per Litre
gm/L gram per Litre
IU International Units
kg Kilogram
M.T Metric Tonnes
mg Milligram
mg/L milligram per litre
nm Nanometer
rpm rotation per minute
µg/kg microgram per kilogram
µg/L microgram per litre