Table Of Content“Prevalence and antibiotic resistance of Listeria monocytogenes from
pork products and piggery environment in Konkan region”
Submitted to
GOA UNIVERSITY
For the award of the degree of
DOCTOR OF PHILOSOPHY IN MICROBIOLOGY
By
Mr. Abhay Vijay Raorane
Work carried out under the guidance of
Dr. S. B. Barbuddhe,
(Guide)
Principal Scientist (Veterinary Public Health)
National Institute of Biotic Stress Management
Raipur, Chattisgarh, India
April, 2015
Certificate
Certified that the research work embodied in this thesis entitled
“Prevalence and antibiotic resistance of Listeria monocytogenes from pork
products and piggery environment in Konkan region” submitted by Mr.
Abhay Vijay Raorane for the award of Doctor of Philosophy degree in
Microbiology at Goa University, Goa, is the original work carried out by the
candidate himself under my supervision and guidance.
Dr. S. B. Barbuddhe, (Guide)
Principal Scientist (Veterinary Public Health)
National Institute of Biotic Stress Management,
Raipur,Chattisgarh, India
Statement
I hereby state that this thesis for Ph.D. degree on “Prevalence and
antibiotic resistance of Listeria monocytogenes from pork products and
piggery environment in Konkan region” 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.
Abhay Vijay Raorane
Ph.D. student
Department of Microbiology
Goa University,
Goa
Acknowledgement
Completion of this doctoral dissertation was possible with the support of
several people. I would like to express my sincere gratitude to all of them. First
of all, I am extremely grateful to my research guide, Dr. S. B. Barbuddhe,
Principal Scientist at National Institute of Biotic Stress Management, ICAR
India, for his valuable guidance, scholarly inputs and consistent encouragement I
received throughout the research work. This feat was possible only because of
the unconditional support provided by Sir. A person with an enthusiastic mind,
most active personality and positive disposition, Sir has always made himself
available to clarify my doubts despite his busy schedules and I consider it as a
great opportunity to do my doctoral programme under his guidance and to learn
from his research expertise.
I thank Dr. N.P. Singh, Director Indian Council of Agriculture Research
Complex for Goa, for the academic support and the facilities provided to carry
out the research work at the Institute.
Dr. S. Bhosale, Professor and Dean, Life science, Goa University, has
been very encouraging and supportive, and I express my gratitude to her.
I owe immensely to my VC’s Nominee, Dr. S. Garg Assistant Professor
Department of Microbiology, Goa University, for his valuable suggestions and
concise comments on the progress report of dissertation.
I am thankful to Dr Z. B. Dubal Senior scientist, IVRI, Izatnagar,
Bareilly and Dr E. B. Chakurkar Principle scientist, ICAR for Goa, for their
imperative help during sample collection program. I express my thanks to
Managing Director of Deonar abattoir for allowing sample collection during pig
slaughtering process. Also thanks to Dr Sudhkar Pawar, he actively helped while
sample collection at slaughterhouse. I thankful to Dr. A. S. Bannalikar, Head,
Department of Veterinary Microbiology, Bombay Veterinary College, and his
students, Dr. Mangesh Hemade and Dr. Swapnil Dalvi for providing laboratory
facilities and hostel accommodation during sample collection programme.
Sample collection was impossible task without active participation of
piggery farmers, my special thanks to Mr Sunil Desai, Mr. Khanolkar, Mr.
Jovian Lewis Mr. Jhon and Father Almeda and their workers for their help in
animal handling while sample collection.
My colleagues, Dr. Swapnil Doijad, Dr. D’çosta, Ms. Sushanta, Ajay,
Supriya, Satyajit, Dr. Shubhada, Krupali, Prathmesh and Mennakshi have all
extended their support in a very special way, and I gained a lot from them,
through their personal and scholarly interactions, their suggestions at various
points of my research programme. I also acknowledge my friends from my
native place and hostel mates, Sukanya, Priti, Sheetal, Susmita, Mitesh, Suyog,
Tushar, Hrishikesh, Amit, Nitin and Chaitanya for their moral and economical
support.
I indebted a lot to my Father and Mother, they scarified all their dreams
and life to provide quality education to us, my elder Sister, Vishakha and
Brother, Ajay who encouraged and helped me at every stage of my personal and
academic life.
Above all, I owe it all to Almighty God for granting me the wisdom,
health and strength to undertake this research task and enabling me to its
completion.
Mr. ABHAY VIJAY RAORANE
Abbreviations
AGP: Antibiotics as Growth Promoters
ALOA: Agar Listeria according to Ottaviani and Agosti
AST: Antibiotic Susceptibility Testing
BAM: Bacteriological Analytical Manual
BHI: Brain Heart infusion (broth)
Bp: Base pairs
CAMP: Christie Atkins Munch Petersons test
CDC: Center of Disease Control
CFU: Colony Forming Unit
CHEF: Contour Clamped Homogeneous Electrophoresis
CLSI: Clinical and Laboratory Standards Institute
DNA: Deoxyribo nucleic acid
dNTP: Deoxyribonucleotide triphosphate
EDTA: Ethylene diamine tetra acetic acid
EP: Efflux Pump
FAO: Food and Agriculture Organization
FDA: Food and Drug Administration
FSIS: Food safety and inspection service
FQ: Fluroquinolones
GMP: Good Manufacturing Practices
HACCP: Hazard Analysis Critical Control Point
ILCC: Indian Listeria Culture Collection
ISO: International Organization of Standardization
Lde: Listeria Drug Efflux pump
L. monocytogenes: Listeria monocytogenes
LLO: Listeriolysin O
mdrL: Mutlidrug resistance (efflux pump) of Listeria species
MFS: Major Facilitator Superfamily
MIC: Minimum Inhibitory Concentration
MLEE Multi Locus Enzyme Electrophoresis
MLST Multi Lcous Sequence typing
MTCC Microbial Type Culture Collection
MTWP Microtiter well plate
OD: Optical Density
PALCAM: Polymyxin Acriflavin Lithium-chloride Aesculin Mannitol (Agar)
PBS: Phosphate Buffered Saline
PC-PLC: Phosphatidylcholine Specific Phospholipase C
PCR: Polymerase chain reaction
PFGE: Pulsed field gel electrophoresis
PI-PLC: Phosphatidylinositol Specific Phospholipase C
QRDR: Quinolone Resistance Determining Region
RNA: Ribose nucleic acid
RTE: Ready-To-Eat (Food)
Spp.: Species
TBE: Tris–borate EDTA buffer
TE: Tris EDTA
TMS: Transmebrane Segments
USDA: United States Department of Agriculture
UVM: University of Vermont media
WHO: World Health Organization
Units of measurement
µg: microgram
µm: micrometer
g: grams
h: hour
kDa: Kilodaltons
M: Molar
mg: miligrams
ml: milliliter
mM: miliMolar
mm: millimeter
O.D.: Optical density
oC: Degree celcius
Pmol: Pico mole
rpm: revolution per minute
µ: micron
Description:Principal Scientist at National Institute of Biotic Stress Management, ICAR. India . Porcine parvo virus infection is the major cause of stillbirth, mummification, rodents, sheep, rabbits, cattle, horses, dogs, and cats also carry pathogenic Y. For preventive care of pigs, antibiotics such as, tet
