Isolation, Antibiotic Resistance, and Molecular Characterization of Campylobacter from Poultry ...

144 Pages·2016·1.16 MB·English

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Doctoral Dissertations Graduate School
12-2012
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Willie James Taylor
University of Tennessee - Knoxville, [email protected]
Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss
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Taylor, Willie James, "Isolation, Antibiotic Resistance, and Molecular Characterization of Campylobacter
from Poultry, Swine and Dairy Cows. " PhD diss., University of Tennessee, 2012.
https://trace.tennessee.edu/utk_graddiss/1566
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To the Graduate Council:
I am submitting herewith a dissertation written by Willie James Taylor entitled "Isolation,
Antibiotic Resistance, and Molecular Characterization of Campylobacter from Poultry, Swine
and Dairy Cows." I have examined the final electronic copy of this dissertation for form and
content and recommend that it be accepted in partial fulfillment of the requirements for the
degree of Doctor of Philosophy, with a major in Food Science and Technology.
F. Ann Draughon, Major Professor
We have read this dissertation and recommend its acceptance:
P. Michael Davidson, Sevetlana Zivanovic, Stephen P. Oliver
Accepted for the Council:
Carolyn R. Hodges
Vice Provost and Dean of the Graduate School
(Original signatures are on file with official student records.)
To the Graduate Council:
I am submitting herewith a dissertation written by Willie James Taylor entitled
“Isolation, Antibiotic Resistance, and Molecular Characterization of
Campylobacter from Poultry, Swine, and Dairy Cows”. I have examined the final
electronic copy of this dissertation for form and content and recommend that it be
accepted in partial fulfillment of the requirements for the degree of Doctor of
Philosophy, with a major in Food Science and Technology

Dr. F. Ann Draughon
Major Professor

We have read this dissertation
and recommend its acceptance:
Dr. Michael Davidson
Dr. Sevetlana Zivanovic
Dr. Stephen Oliver

Accepted for the Council:
Carolyn R. Hodges
Vice Provost and Dean of the Graduate School

(Original signatures are on file with official student records.)
Isolation, Antibiotic Resistance, and Molecular
Characterization of Campylobacter from Poultry,
Swine and Dairy Cows

A Dissertation
Presented for the
Doctor of Philosophy
Degree
The University of Tennessee, Knoxville

Willie James Taylor
December, 2012
Dedication
I would like dedicate this dissertation to my wife (Melissa), my mother
(Lois), my step-mother (Mrs. B), my family (Sue, Bae, Lee, and Tamika), my-
step-dad (Bernard), my dad (Willis), my son (Willie) and his mother (Helen).

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Acknowledgements
I would like to thank Dr. Ann Draughon, for without her belief in my
abilities, I would not be where I am today. I would also like to thank my
committee members, Dr. Michael P. Davidson, Dr. Svetlana Zivanovic, and Dr.
Stephen P. Oliver for their continued support and patience over the past three
years and throughout this entire process.
Special thanks go to my wife (Melissa) for her patience and sacrifice
throughout this entire process. Without her support, persistence, and
understanding, I could not have completed this dissertation. This
accomplishment is due to her. Thanks.
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Abstract
This study was conducted to determine presence, antibiotic resistance,
and genetic relatedness of Campylobacter on diverse farm types. Cloacal fecal
samples from poultry (broilers, layers, and turkey breeders), and fecal rectal
samples from non-poultry animals (farrowing sows, finisher pigs, and lactating
dairy cows) were tested for Campylobacter using BAM protocols. Agar disk
diffusion method was used to determine antibiotic resistance, and PFGE analysis
to determine genetic relatedness of isolates recovered within the different farm
types. Campylobacter was detected in 34.8% (range of 6.7 to 62.0%, P < 0.001)
of cloacal samples from poultry, in 61.4% (range of 55.0 to 69.0%, P > 0.05), and
26.0% (range of 19.0 to 34.0%, P < 0.05) of rectal samples from pigs, and dairy
cows, respectively. Antibiotic resistance was detected in 30.2% (range of 0 to
82.4%, P < 0.001) of Campylobacter from poultry, in 54.7% (range of 24.2 to
100%, P < 0.001), and 14.3% (range of 0 to 28.6%, P < 0.05) of Campylobacter
from pigs and dairy cows, respectively. Isolates from poultry with resistance
against ciprofloxacin (range of 0 to 79.4%, P < 0.001) were detected more often
than those with resistance to erythromycin (range of 0 to 8.8%, P < 0.01) or
gentamicin (range of 0 to 4.5%, P > 0.05). Isolates from pigs with resistance to
erythromycin (range of 20.0 to 100%, P < 0.001) were detected more often than
those with resistance to ciprofloxacin (range of 0 to 11.4%, P < 0.05) or
gentamicin (2.6%, range of 0 to 3.0%, P > 0.05). Resistance to ciprofloxacin
(range of 0 to 7.7%, P < 0.05), erythromycin (range of 0 to 28.6%, P < 0.01), and
gentamicin (range of 0 to 18.8%, P < 0.01) varied between the different dairy
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farms. Genotyping by PFGE showed that Campylobacter strain types recovered
from different poultry and non-poultry production types had distinctly different
PFGE patterns. In conclusion, marked disparities existed within different poultry
and non-poultry production types in terms of Campylobacter presence, antibiotic
resistance, and strain types.

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Table of Contents
Introduction..................................................................................................1
References ...........................................................................................3
Part I: Literature Review................................................................................5
Campylobacter General Characteristics...............................................6
Bacteriological............................................................................6
Physiological..............................................................................6
Survival......................................................................................7
Clinical Significance............................................................................11
Human Campylobacteriosis.....................................................12
Sequelæ...................................................................................13
Epidemiology......................................................................................14
Mode of Transmission of Campylobacter to Food Animals.................16
Vertical.....................................................................................18
Horizontal.................................................................................20
Campylobacter Contamination at Slaughter.......................................30
Poultry......................................................................................30
Pork..........................................................................................33
Campylobacter Colonization in Broilers..............................................34
Campylobacter Colonization in Layers ...............................................35
Campylobacter Colonization in Turkeys..............................................36
Campylobacter Colonization in Swine ................................................37
Campylobacter Colonization in Dairy Cows........................................38
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Campylobacter from Diverse Production Types..................................38
Antibiotic Resistance in Campylobacter..............................................41
Types of Antibiotic Resistance.................................................41
Antibiotics in Treatment of Campylobacter Infections..............44
Antibiotic Susceptibility Testing Methods............................................48
Campylobacter Resistance.................................................................50
Resistant Campylobacter on Diverse Production Types.....................53
Pulsed Field Gel Electrophoresis........................................................54
References .........................................................................................55
Part II: Isolation, Antibiotic Resistance and Molecular Characterization of
Campylobacter from Poultry in Different Types of Poultry Productions........68
Abstract...............................................................................................69
Introduction.........................................................................................70
Materials and Methods........................................................................71
Farm Selection.........................................................................71
Sample Collection....................................................................72
Sample Preparation and Selective Enrichment........................72
Campylobacter spp. Determination..........................................72
Antibiotic Resistance Testing...................................................73
PFGE Analysis.........................................................................74
Statistical Analysis...................................................................76
Results and Discussion.......................................................................76
Campylobacter Isolation...........................................................76
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