THESIS Submitted in partial fulfillment of requirements for the Degree of ...

140 Pages·2015·1.79 MB·English

MOLECULAR TYPES AND ANTIMICROBIAL RESISTANCE PROFILES
OF Staphylococcus aureus FROM BOVINE MASTITIS

T H E S I S
Submitted
in partial fulfillment of requirements for the Degree of

M A S T E R O F V E T E R I N A R Y S C I E N C E

IN
VETERINARY PUBLIC HEALTH

BY

GANDHALE DATTATRAY GANPAT
Enrolment No: V/02/354

Krantisinh Nana Patil College of Veterinary Science, Shirwal

MAHARASHTRA ANIMAL AND FISHERY SCIENCES
UNIVERSITY, NAGPUR- 440 001
(INDIA)
2015
DECLARATION OF STUDENT

I hereby declare that the experimental Research work and
interpretation of the thesis entitled MOLECULAR TYPES AND
ANTIMICROBIAL RESISTANCE PROFILES OF Staphylococcus aureus FROM
BOVINE MASTITIS or part thereof has not been submitted for any other
degree or diploma of any University, nor the data have been derived from
any thesis/publication of any University or scientific organization. The
sources of materials used and all assistance received during the course of
investigation have been duly acknowledged.

Date : Signature

(GANDHALE DATTATRAY GANPAT)
Enrolment No: V/02/354

Counter signed by Chairman,
Advisory Committee with date
DECLARATION OF ADVISORY COMMITTEE

Shri. GANDHALE DATTATRAY GANPAT has satisfactorily
prosecuted his course of research for a period of not less than one
semester and that the thesis entitled, MOLECULAR TYPES AND
ANTIMICROBIAL RESISTANCE PROFILES OF Staphylococcus aureus FROM
BOVINE MASTITIS submitted by him is the result of research work is
sufficient to warrant its presentation to the examination in the subject of
VETERINARY PUBLIC HEALTH for the award of MASTER OF
VETERINARY SCIENCE degree by the Maharashtra Animal and Fishery
Sciences University, Nagpur.

We also certify that the thesis or part thereof has not been previously
submitted by her for a degree of any other University.

Place : Shirwal
(Dr. R. P. KOLHE)
Date : Advisor/Guide
Assistant Professor

Advisory Committee
Name and Designation Signature

1) Dr. R.P. Kolhe, Chairman
Assistant Professor, Dept. of Veterinary Public Health

2) Dr. P.D. Deshpande, Member
Associate Professor, Dept. of Veterinary Public Health

3) Dr. D. M. Muglikar, Member
Associate Professor, Dept. of Veterinary Microbiology

4) Dr. S. N. Jadhav, Member
Assistant Professor, Dept. of Veterinary Biochemistry
CERTIFICATE

This is to certify that the thesis entitled, MOLECULAR TYPES AND
ANTIMICROBIAL RESISTANCE PROFILES OF Staphylococcus aureus
FROM BOVINE MASTITIS submitted by GANDHALE DATTATRAY
GANPAT to the Maharashtra Animal and Fishery Sciences University in
partial fulfillment of the requirement for the degree of MASTER OF
VETERINARY SCIENCE has been approved by the Student's Advisory
Committee after examination in collaboration with the External Examiner.

Name & Signature of Signature with Seal Name & Signature
External Examiner Head of Department Advisor/Guide

Advisory Committee

Name and Designation Signature

2) Dr. R.P. Kolhe, Chairman
Assistant Professor, Dept. of Veterinary Public Health

2) Dr. P.D. Deshpande, Member
Associate Professor, Dept. of Veterinary Public Health

3) Dr. D. M. Muglikar, Member
Associate Professor, Dept. of Veterinary Microbiology

4) Dr. S. N. Jadhav, Member
Assistant Professor, Dept. of Veterinary Biochemistry

Signature with seal
Dean/Associate Dean
TABLE OF CONTENTS

Chapter Page

I) INTRODUCTION 1 - 4

II) REVIEW OF LITERATURE 5 - 33

III) MATERIALS & METHODS 34 - 42

IV) RESULTS & DISCUSSION 43 - 62

V) SUMMARY & CONCLUSION(S) 63 - 66

A) BIBLIOGRAPHY i - xiii

B) APPENDIX AND ANNEXURE xiv - xxv

C) VITA xxvi
LIST OF TABLES

Page No/
Table No. Title of the table Between
Page
1 Details of samples collected during study 35-36
2 Biochemical characteristics of S. aureus. 36
3 Oligonucleotide sequences used for detection of S. aureus 39-40
enterotoxin genes.
4 Antimicrobial agents used and their concentrations 41
5 Oligonucleotide sequences used for detection of antibiotic 41
resistance genes in S. aureus
6 Prevalence of subclinical and clinical mastitis at different farms 44
7 Isolation of S. aureus from different samples 47
8 Farm wise prevalence of S. aureus isolates 48-49
9 Farm wise distribution of enterotoxin types of S. aureus isolates 49 - 50
10 Antimicrobial resistance pattern of S. aureus by disk diffusion 55- 56
method
11 Farm wise genotypic antimicrobial resistance pattern of S. aureus 60 - 61
isolates
12 Isolate wise details of S. aureus enterotoxin 62 – 63
13 Isolate wise phenotypic and genotypic antimicrobial resistance 62 – 63
pattern of S. aureus
14 Farm wise antimicrobial resistance pattern of S. aureus isolates 62 – 63
15 Dairy management practices followed at different farms 62 – 63
16 Teat wise prevalence of subclinical mastitis at different farms 62 – 63
LIST OF FIGURES

Fig No. Title Page No.

1 Colony characteristics of S. aureus on Mannitol Salt Agar A

2 Antibiotic sensitivity/ resistance pattern of S. aureus on Muller B
Hinton Agar

3 S. aureus enterotoxin genes (set A) C

4 S. aureus enterotoxin genes (set B) D

5 Detection of antimicrobial resistance genes in S. aureus isolates E

6 Farm wise prevalence of subclinical and clinical mastitis F

7 Prevalence of S. aureus from different sources of samples F

8 Farm wise prevalence of S. aureus from mastitis milk G

9 Antimicrobial resistance pattern of S. aureus isolates G

10 Distribution of antimicrobial genes in S. aureus isolates H
DEDICATED TO MY FAMILY AND
FRIENDS
Acknowledgement
I do not have exact words to express my deepest and sincere
feelings of thanks towards Dr. R. P. Kolhe, my Major Advisor and
Assistant professor, Department of Veterinary Public Health, KNP
College of veterinary Science, Shirwal. The whole credit of research
work done by me goes to him for his valuable guidance, hard work,
dedication, patience, encouragement. This thesis would not have been
possible without the help, support and understanding of him.
I take this opportunity to express my deep sense of gratitude to
my member of advisory committee and Head, Department of
veterinary Public Health Dr. P. D. Deshpandefor hisconsistent
inspirations, invaluable guidance, cooperation, and understanding
throughout this study.
I wish to express my sincere thanks to Dr. D. M. Muglikar and
Dr. S. N. Jadhav, members of advisory committee for their valuable
suggestions and technical support for completion of this study. I
respectfully thanks to Dr. P. P. Mhase, Assistant professor,
Department of Microbiology, KNPVC, Shirwal, for his support. I am
indebted to Dr. H. S. Birade, Associate Dean, PGI, Akola, Dr. S. B.
Kawitkar, Dr. SmitaSurkar, Dr. B. R. Kadam andDr. T. C. Shende for
providing me valuable help and advice during the course this study. I
also never forget the motivation and support of my friends Engineer
ManojGosavi and Dr. A. M. Jadhav.
My special thanks goes to Dr. A. H. Ulemale, In charge, CIF for
providing every possible facility for this research work.
I would like to pay my sincere thanks to Dr. A.S. Bannalikar,
Associate Dean of the college for support and facilities for this thesis
work and also I also thanks to S. V. Mote, for his timely help.
I would like to give sincere thanks to Dr. A. K. Misra,Honorable
Vice Chancellor of Maharashtra Animal & Fishery Sciences University
Nagpur for providing me opportunity after a long period for higher
education as an in service candidate by sanctioning my study leave. I
would also like to acknowledge the help of Dr. S. T. Bapat, former
Associate Dean, KNP College of veterinary science, Shirwal because of
whom it was possible for me to accomplish this task.
The words are insufficient to express my feelings about my
loving father and mother, brother Sunil and sisterBharati, without their
hard work and support it is not possible to me to reach this destination.
I thank to God for bringing in my life loving wife Mayuri and beloved
daughter Ishwari, with their dedication, continuous support and
motivation,I am able to complete this work.
I extent my thanks to Saba Nalband, UrmilaJagtap,
ChandrakantDhandore, JaishriRohakale, SujataBhave, Dr. D. M.
Khandagale and TejalWalunj for their moral help. I thank to God for
giving me precious gift in the form of friends Satish, Datta, Digamber,
Somanath, Nitin, Kaustubh,Yogesh who always supported me.
I am very much thank full to all KNP college staff for their co-
operation and motivation throughout the course of study. I also thanks
all those who have helped me directly or indirectly in completion of
this study.
I sincerely thanks to the God who made everything possible in my life.
I wish to dedicate this thesis to my beloved family and friends.
Place: Shirwal
Date: 04/ 07/ 2015
(GandhaleDattatrayGanpat)
En. No. V/02/354