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Evaluation of the efficacy of two different tulathromycin treatments in weaned piglets infected intratracheally with Haemophilus parasuis serovar 5 [Elektronische Ressource] / Rose-Leah Kavita Austin-Busse

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From the Clinic for Swine (Head: Prof. Dr. Dr. habil. Karl Heinritzi) of the Veterinary Faculty of the Ludwig-Maximilians-University Munich and the Clinic for Swine (Head: Prof. Dr. Mathias Ritzmann) of the Veterinary University Vienna Evaluation of the efficacy of two different tulathromycin treatments in weaned piglets infected intratracheally with Haemophilus parasuis serovar 5 Inaugural-Dissertation for the veterinary doctorate of the Veterinary Faculty of the Ludwig-Maximilians-University Munich Rose-Leah Kavita Austin-Busse Georgetown, Guyana Munich 2010 Printed with permission from the Veterinary Faculty of the Ludwig-Maximilians-University Munich Dean: Univ.-Prof. Dr. Joachim Braun Referent: Univ.-Prof. Dr. Dr. habil. Karl Heinritzi Korreferent: Day of Promotion: th24 July 2010 EmmanuelContents I Contents Contents..................................................................................................................... I Abbreviations............................................................................................................ V 1 Introduction ........................................................................................................ 1 2 Literature review ................................................................................................ 3 2.

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Published 01 January 2010
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From the Clinic for Swine
(Head: Prof. Dr. Dr. habil. Karl Heinritzi)
of the Veterinary Faculty
of the Ludwig-Maximilians-University Munich
and the
Clinic for Swine
(Head: Prof. Dr. Mathias Ritzmann)
of the Veterinary University Vienna






Evaluation of the efficacy of two different tulathromycin
treatments in weaned piglets infected intratracheally with
Haemophilus parasuis serovar 5




Inaugural-Dissertation
for the veterinary doctorate
of the Veterinary Faculty
of the Ludwig-Maximilians-University Munich

Rose-Leah Kavita Austin-Busse
Georgetown, Guyana


Munich 2010
Printed with permission from the Veterinary Faculty
of the Ludwig-Maximilians-University Munich











Dean: Univ.-Prof. Dr. Joachim Braun
Referent: Univ.-Prof. Dr. Dr. habil. Karl Heinritzi
Korreferent:















Day of Promotion:
th24 July 2010















EmmanuelContents I
Contents
Contents..................................................................................................................... I
Abbreviations............................................................................................................ V
1 Introduction ........................................................................................................ 1
2 Literature review ................................................................................................ 3
2.1 Haemophilus parasuis ................................................................................ 3
2.1.1 Etiology ................................................................................................... 3
2.1.2 Epidemiology........................................................................................... 5
2.1.3 Experimental infection............................................................................. 9
2.1.4 Pathogenesis and virulence .................................................................. 12
2.1.5 Clinical signs ......................................................................................... 15
2.1.6 Gross and microscopic lesions.............................................................. 17
2.1.7 Differential diagnosis............................................................................. 18
2.2 Diagnosis ................................................................................................... 19
2.2.1 Selection of animals and sampling sites................................................ 19
2.2.2 Sampling procedures ............................................................................ 20
2.2.3 Bacterial isolation .................................................................................. 20
2.2.4 Molecular biological methods ................................................................ 21
2.2.5 Further diagnostic methods................................................................... 23
2.3 Therapy ...................................................................................................... 25
2.3.1 Antimicrobial use in food animal production .......................................... 25
2.3.2 Antimicrobial treatment for HPS ............................................................ 25
2.3.3 Antimicrobial susceptibility patterns for HPS ......................................... 26
2.3.4 Tulathromycin........................................................................................ 27
2.3.5 Prophylaxis and immunity ..................................................................... 29
3 Material and Methods ...................................................................................... 32
3.1 Study aim ................................................................................................... 32
3.2 Study animals ............................................................................................ 32
3.3 Study design.............................................................................................. 33
3.3.1 Housing ................................................................................................. 34
3.3.2 Biosecurity............................................................................................. 36 Contents II
3.3.3 Administration of tulathromycin ............................................................. 36
3.3.4 Preparation of challenge inoculum ........................................................ 36
3.3.5 Administration of inoculum .................................................................... 37
3.3.6 Euthansia .............................................................................................. 38
3.4 Clinical examinations................................................................................ 38
3.5 Necropsy.................................................................................................... 40
3.6 Sample collection and Diagnostic ........................................................... 42
3.6.1 Blood sampling...................................................................................... 42
3.6.2 Synovial and cerebrospinal fluid collection............................................ 43
3.6.3 Collection and processing of serosal swabs.......................................... 43
3.6.4 Bacteriological culture ........................................................................... 44
3.6.5 Histopathological examination............................................................... 44
3.6.6 Polymerase chain reaction (PCR) analysis for HPS.............................. 45
3.7 Statistics .................................................................................................... 45
4 Results.............................................................................................................. 47
4.1 Screening, animal numbers and data collection .................................... 47
4.2 Clinical results........................................................................................... 48
4.2.1 Mortalities.............................................................................................. 48
4.2.2 Clinical examinations............................................................................. 49
4.2.3 Average daily gain (ADG)...................................................................... 51
4.2.4 Rectal temperatures.............................................................................. 52
4.2.5 Blood examination (Leukocyte population)............................................ 53
4.3 Gross lesions............................................................................................. 55
4.3.1 Lung assessment .................................................................................. 55
4.3.2 Synovial fluid assessment ..................................................................... 56
4.3.3 Cerebrospinal fluid assessment ............................................................ 57
4.3.4 Brain assessment.................................................................................. 58
4.3.5 Assessment of the serosal membranes and cavities............................. 60
4.4 Histopathological lesions ......................................................................... 64
4.5 PCR detection of HPS genome................................................................. 65
4.6 Bacterial isolation of HPS......................................................................... 66
4.7 Association between HPS detection in collective serosal swabs by PCR
and bacterial culture ................................................................................. 67 Contents III
4.8 Associations between HPS detection in samples from challenged pigs
and clinical results.................................................................................... 67
4.8.1 Association between HPS detection by PCR and bacterial culture and
the median total daily clinical score....................................................... 67
4.8.2 Association between HPS detection by PCR and bacterial culture and
time of death.......................................................................................... 68
4.9 Association between HPS detection in samples from challenged pigs
and gross lesions...................................................................................... 70
4.9.1 Association between HPS detection in collective serosal swabs and the
total inflammation score ........................................................................ 70
4.9.2 Association between HPS detection in collective serosal swabs and
pleuritis, pericarditis, and peritonitis ...................................................... 71
4.9.3 Association between HPS genome detection in cerebrospinal fluids,
brain swabs and the assessment of the brains and cerebrospinal fluids73
4.9.4 Association between HPS genome detection in synovial fluids, joint
capsules and the synovial fluid assessment.......................................... 76
4.10 Association between HPS detection in samples from challenged pigs
and histopathological lesions.................................................................. 77
4.10.1 Association between HPS genome detection in synovial fluids and joint
capsules and synovitis .......................................................................... 77
4.10.2 Association between HPS genome detection in cerebrospinal fluids and
brain swabs and meningitis ................................................................... 78
4.10.3 Association between HPS detection and pyelitis................................... 79
5 Discussion........................................................................................................ 80
5.1 Clinical results........................................................................................... 80
5.1.1 Mortalities.............................................................................................. 80
5.1.2 Clinical examinations............................................................................. 81
5.1.3 Average daily gain (ADG)...................................................................... 84
5.1.4 Rectal temperatures.............................................................................. 84
5.2 Blood examination (Leukocyte population) ............................................ 85
5.3 Gross lesions............................................................................................. 86
5.3.1 Lung lesions .......................................................................................... 86
5.3.2 Synovial fluid assessment ..................................................................... 88
5.3.3 Cerebrospinal fluid assessment ............................................................ 89 Contents IV
5.3.4 Brain assessment.................................................................................. 89
5.3.5 Assessment of serosal membranes and cavities................................... 90
5.4 Histopathological lesions ......................................................................... 91
5.5 Detection of HPS by PCR and bacterial culture...................................... 92
5.6 Comparison of HPS detection by PCR and bacterial culture in collective
serosal swabs from challenged pigs....................................................... 93
5.7 Associations between HPS detection, the clinical results and
pathological lesions.................................................................................. 94
5.7.1 Association between HPS detection and clinical results ....................... 94
5.7.2 Association between HPS detection and pathological lesions .............. 95
5.8 Conclusion................................................................................................. 97
6 Summary........................................................................................................... 99
7 Zusammenfassung ........................................................................................ 101
8 List of Figures ................................................................................................ 103
9 List of Tables.................................................................................................. 105
10 References...................................................................................................... 107
11 Acknowledgements ....................................................................................... 123 Abbreviations V
Abbreviations

ADG Average daily gain
AGPT Agar gel precipitation test
BALF Bronchoalveolar lavage fluid
BW Body weight
CD Colostrum-deprived
CDCD Caesarean derived-colostrum-deprived
CFU Colony forming units
CNS Central nervous system
DIC Disseminated intravascular coagulation
ELISA Enzyme linked immunosorbent assays
ERIC-PCR Enterobacterial repetitive intergenic consensus
polymerase chain reaction
HE Haematoxlylin and eosin
HEPA High efficiency particulate arresting filter
HPS Haemophilus parasuis
IHC Immunohistochemistry
IHA Indirect haemagglutination
IM Intramuscular
ISH In situ hybridisation
KDa Kilodalton
MIC Minimum inhibitory concentration
NAD Nicotine adenine dinucleotide
NADH Nicotine adenine dinucleotide plus hydrogen
PBS Phosphate buffered saline
PCR Polymerase chain reaction
PCV-2 Porcine circovirus type 2
PMWS Post weaning multisystemic wasting syndrome
PPLO Pleuropneumonia like organism
PRRSV Porcine reproductive and respiratory syndrome virus
rep-PCR Repetitive element-based polymerase chain reaction
RFLP Restriction fragment length polymorphism Abbreviations VI
SDS-PAGE Sodium dodecyl sulfate polyacrimde gel eletrophorese
SEW Segregated early weaning
SPF Specific pathogen free
SRDC Swine respiratory disease complex