Campylobacter abortion in sheep

257 Pages·2010·6.83 MB·English

by  Mannering, Sally Anne

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Campylobacter abortion in sheep:
a study of strain types and vaccine protection.
A thespirse senitnef dul filmoefnt th er equiremfeonrtt hsed egreoef
DoctoorfP hilosoipnhV ye terinPaartyh ology
atM assey UnivPearslimtye,r sNtoornt Nhe,w Z ealand.
SallAyn neM annering
2003
Abstract
The bacteria Campylobacter fetus subsp. fetus and C. jejuni are important causes of
abortion in sheep, and the majority of Campylobacter sheep abortion in New Zealand is
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associated with C. fetus subsp. fetus. Campylovexin (Schering-Plough Animal Health
Ltd., Upper Hutt) is a single-strain, killed vaccine available in New Zealand for the
prevention of abortion in sheep due to C. fetus subsp. fetus. The present study was
initiated in response to questions regarding the protection offered by this vaccine
against other strains of C. fetus subsp. fetus causing sheep abortion in New Zealand.
Campylobacter isolates from aborted sheep foetuses were collected after culture in
veterinary diagnostic laboratories from sheep flocks in the Hawke's Bay region in 1999.
This was extended in 2000, when Campylobacter sheep abortion isolates cultured in the
same way from farms throughout New Zealand were collected. In total, 374 C. fetus
subsp. fetus isolates, 30 C. jejuni isolates, and one C. coli isolate were collected from
249 fanus. These isolates were sub-typed using the genetic technique, pulsed-field gel
electrophoresis (pFGE). A total of 26 distinct PFGE types were identified amongst the
C. fetus subsp. fetus sheep abortion isolates, and one type, PFGE type B 1,
predominated. Of the farms from which C. fetus subsp. fetus was isolated, PFGE type
B1 was found on 19 out of the 25 fanus in the Hawke's Bay study (75%) and 132 out of
the 200 farms in the national study (66%).
Nine distinct PFGE types were found amongst the 30 C. jejuni isolates from the
Hawke's Bay and national studies. No single C. jejuni PFGE type was identified from
sheep abortion most frequently. Investigation of a large sheep abortion outbreak initially
attributed to C. jejuni revealed that the initial diagnosis was misleading due to
contamination of the foetal sample, and that the outbreak was more likely to have been
associated with C.fetus subsp.fetus.
Seven C. fetus subsp. fetus isolates representative of the genotypes found amongst New
Zealand sheep abortion isolates in a 1987 study were lodged with the New Zealand
Reference Culture Collection, Medical Section (de Lisle et al. 1987). These isolates
were typed in the present study using PFGE, and were found to be similar or
11
indistingufriosmh tahbelP eFG E typesi dentiifinet dh eH awke'Bsa yo rn ational
studieTsh.e fr equenocfyi solatoifto hnet ypeasl saop pearetdob es imilbare tween
1987 and 2000, ast hper evalteynpfrteo m1 987 wasi dentiafisPe FdG E typBe 1 .
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A nationwciadlfelo rs uspecctaesde osfC ampylovexibnreakdowrne sulted in the
identificoafot nieofl no ckin w hicahb ortidounets oC. fetus subsfpet.us werree ported
ine wesa pparentvlayc cinattheadt season. Ap4p0r eowxeisam baotretloeyud to fa
flocko f1 100 ewesa,n di tw ast hougthhtaf to ure wetsh ahta da bortweedr vea cicnated
thaste asoTnh.eh ighs tockdienngs imtanya gemenotft hiflso ckm eantth atth erwea s
liketloyh avebe ena high chaltlove ancgcei naetweedsfr om unvaccinaabtoertdi ng
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ewesI.n vestigoaftt iwooon t hearp parenCtam pylovexibnreakdowcna serse vealed
thaetw esh adn otb eenv accinattheasdte asoannd werea lsruno ata highs tocking
densiInt ya.d dititohneC., f etus sUbs.pfetus isolaitnthe ess et woc asewse rteh seam e
PFGE typaes t hev accisntrea iPnF,G E typAeI ,w hicshu ggesttheadit nt hescea ses
apparetn vaccibnree akdowwans n otd uet ol acokf c ross-protaegcatiinoostnth er
strsa.iD nespittheew idesproecacdur renocfde i stisntrcaitn so fC. fetus subsfpet.us ,
abortidoune t oC. fetus subsfpet.us ine wesv accinawtietCdha mpylov®e axpipneared
tob er area ndw asn oats sociwaitteahdp articuPlFGaEr t ype.
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Serum from 15 ewes vacciwniatht Ceamdp ylovexiwnasc ollecantde du sedi n
Westebrnl otteixnpge rimetnoti sn vestiangtaitbeo dbyi ndiwnigth th e proteintsh eo f
variouPsFG E typeosfC. fetus subsfpet.us . Serumfr om vaccinaetweedsc ontained
antibodtiheasbt o untdo t hep roteoifnt sh ed iffereC.n ftet us subsfpet.us PFGE types.
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Thisin vitro studsyu ggesttheadst h eevpa ccinated with Camdpeyvleolvoepx in
antibotdhiaertse cognipsreo teoifen as cohf t hC.e fetus subsfpet.us straitny pes.
Vaccicnhea lleenxgpee rimweenrtpese rformuesdi npgr egnangtu inpeiag ass a model.
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Initiailtwl ays,s hownu sintgh imso detlh aCtam pylovexipnrovidae sdt atistically
significlanetv oeflp rotecfrtoimoh no mologcohuasl lewnigteth h vea ccisntrea iTnh.e n,
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usinang isolaotfPe F GE typBel , itw asd emonstrattheadCt am pylovexianlso
providase tda tistically siognfpi rfioctanetc atlgieaovinenh lse tt erolocghoaulsl enge.
Despithtee r angoefC. fetus susbpf.et us PFGE typeisd entiifinte hdi sst udtyh,e rwea s
tr @
noe videntcose u ggetshta tth ed ifferseanitnt ypecso mpromiCsaemdp ylovexin
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efficacyi np rotectaigoainn sC.t fetsuusb sfp.e tsuhse ep abortiporno.t eTihne
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recognitbiyos ne rufrmo me wesv accinawtiethCd am pylovex,it nhel acokf a robust
vaccibnree akdowcnas e,an d thed emosntratoifoc nr oss-protientc htegi uoinn epai g
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modelp,r oviedvei dentcheaC tam pylovexaipnpeartso o ffecrr oss-cptriooatneg ainst
straionfsC. fetussu bsfpet.u csa usianbgo rtiinos nh eeipnN ewZ ealand.
IV
Acknowledgements
Ith ank my team of supervisors:
Dr. Dave West Associate Professor
Institute of Veterinary, Animal and Biomedical Sciences,
Massey University, Palmerston North
Dr. Stan Fenwick Senior Lecturer in Veterinary Public Health
School of Veterinary Clinical Science, Murdoch University,
Perth, Australia
Dr. Roger Marchant Veterinary Adviser
Schering-Plough Animal Health Limited, Upper Hurt
Dr. Kevin O'Connell Technical Manager R&D Global Vaccine Unit
Schering-Plough Animal Health Limited, Upper Hurt
It hank Stan and Roger for dreaming up this project, constructing a plan and obtaining
the funding from Technology New Zealand. Iw ould like to thank them both for their
support and especially Roger for his endless enthusiasm. Dave has been a fabulous
strength in this project and It hank him for his support and for getting me to think.
Trudy Owen (Regulatory Affairs Manager, SPAH ) has been an invaluable source of
advice and support and provided useful input at the project meetings. Iw ould also like
to recognise the time and support of Marty PhiIlips (Division Manager, SP AH) towards
this project.
It hank the friendly crowd at SPAH for making me feel welcome, particularly the staff
of R&D, QA, and QC. Iw ould especially like to thank Phil ip Wallace (Wally) and
Ja cquie Clarke for providing a home-away-from-home for me in the Hurt.
It hank Megan Leyland (pFGE Typing Laboratory technician, IV ABS) for sharing her
considerable PFGE expertise, including help with Diversity Database and with the
production of endless dendrograms. Iw ould like to thank the microbiology staff of the
veterinary diagnostic laboratories who supplied Campylobacter abortion isolates. I
thank Kylie Walker (Micro Lab technician, IV ABS) for storing the Campylobacter
v
isolafrtoems th e Hawke'Bsa yi n1 999 andt hes taaftfV eterinSaerrvyi ceHsa wke's
BayL tdf.o thre icro ntinsuuepdp oorftt h ipsr oject.
Thanky out ot hvee terinarwihanoas d visuesdo fs uspecbtreedakthr oughc ases:
SonyJaa cks(oNno rtCha nterbVuertye rinary SaenrdCv aricoelsynH) am il(lE dendale
VetC luba)n,d t hfearm ersf othre icro operaitnit ohnii sn vestigation.
IV
Iw oulldi kteot hank ABS vetreinarianDsr sA.n neRi dlerM,ar k Owen,Da veW est,
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AlisQouni nAnl,a stJaoihrn stMoarnk, C olleantdt e,s peciDarlK layt hPyar ton ABS
LaboratAonriym aVle terinarfioanar)d ,v iancde assistwanictegh u inepai gp rocedures
ofa veterinnarayt ureI.th ank DebbiCeh esterfi(eSlmda lAnli malP roductUinoint ,
MasseUyn iversanidt Ryo)b H azelwoo(dS AHP AnimalB reediUnngi tf)o ard viancde
IV
assistanIc weo.u ldl ikteo th ank Dr NigePle rki(nEsp idemiologAiBsSt),f or
sttaistiacdavli caned Dr Alan Murrayf or helspufuglg estiIoth nasn.k D r Anne
Midwintfeorras sistanwciet hth e Petroff-Hausseran dc oMuinkteHe org an (post­
IV
mortermo omt echniciAanB,S )f oars sistance.
Finalbluyt,n otl easItw ,o ulldi kteo t hanmky familyan d friendfso rt heirs upport,
particulAanrnleyR idleJru,l iTeh ompsoannd, RowenaT ayolrf orr eveiwintgh esis
drafts.
Thisw orkw ass upportbye da TechnoloNgeyw ZealanTde chnolofgoyrI ndustry
Fellowsinh ciopn juncwtiitoShnc hering-PAlnoiumgaHhle alLtthd .
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TabloefC otnents
Abstract 11
Acknolwedgments v
Tabloef c ontents V11
Lisotf fi gures x
Lisotf t ables xv
Lisotf a ppednices XVI11
Chapt1e:Lr i terarteuvireew 1
1.I1n troduction 2
12. Thgee nCuasmp ylobacter 2
13. IdentifiocfCaa mtpyiloobnac ter species 5
14. EpidemioofCla ompgylyob acter abortinis ohne ep 14
15. Stratiynp ionfCga m pylobacter sheaebpo ritsioolna tes 25
16.P reventCiamopnyl oboacfte r abortini sohnese p 32
17. Surflaacyepe rro t(eSiLnPss ) 36
18. Animal challmeondgeefl osC.r f etus subfsetpus. v acceifnfiec acy 45
19. Objectoiftv hpeers e ssetnutd y 48
Chapt2: eGre neMraatle rainaMdle st hdos 50
2.M1a nufactaudrderress's es 51
2.M2e disat,a ainnsdso lutions 52
2.C3a mpylobacter growtahn idd entification 55
2.P4u lsedg-efielel ledc tro(pphFoGrEe)s is 59
2.S5D S-polyacgreyelll ameicdtreo phoresis 61
2.C6o lleocfts ieornfru om mv accineawteesd 62
2.W7e stbelrno tting 63
VII
Chapt3e:Tr y pionfCg a mpylobacfteteurss u bfsetpuform
. s
sheaebpo rtuisoinpnsug l sedg-efielel ledc torroep:sh ipasi lot
stuodfHy a wk'eBsa yi solates 65
3.I1n troduction 66
3.M2a terainamdle st hods 68
3.R3e sults 70
34. Discussion 75
3.C5o nclusions 77
Chaeprt4 :T ypionfCg a mpyolbacfteteurss u bfsetpussh eep
.
abortiisootnleu ass ipnugl sedg-efielel ledc troip:sh Noaer we s
Zealasntdu dy 78
4.I1n trtoidounc 79
4.M2a terainamdle st hods 80
4.R3e sults 83
4.D4 iscussion 94
4.C5o nclusions 97
Chpate5r:P ulesd-fieledl ergcoetplh oroefCs amipysl boacter
jejunsihe aebpo rtiisootnle as 98
5.I1n troduction 99
5.M2a terainamdle sth ods 101
5.R3e uslts 103
5.D4i scuns sio 101
5.C5o nclusions 151
viii
Chapte6r:I n vitro proteainnd s erologsitcuadlio efs
Campylobacter fetus subsfpet.u s strains 161
61. Introduction 171
6.M2a terainamdle st hods 181
6.R3e sults 132
6.D4i scussion 132
6.C5o nclusions 153
@
Chapte7:r Campylovexbirneakdoiwnn vestigation1 36
71.I ntroduction 137
72. Materanidam lest hods 139
7.R3e sults 141
7.D4i scussion 174
7.C5o nclusions 151
@
Chapte8:r Campylovexeifnficaicnyt hep regnagnuti nea
pigm odel 125
8.I1n troduction 135
8.G2e neMraatle ranidaM lesth ods 155
8.D3e terminoafMADt io son f oArl andB I 158
8.V4a cicnceh alleexnpgmeeer nits 177
8.C5o nclusions 109
9:
Chapter Generdails cussion 191
91. Introduction 129
9.S2tr atiynp iofnCagm pylobacterf etus sbusp.Jetus 129
9.C3a mpylobacter jejuni 149
9.I4n v itro protaenisdne rolosgitcuadli es 159
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9.C5am pylovebxrienak doiwnnv estigation 169
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9.I6n vestiogfCaatmipoynl ovperxoitneucstiinoganinm al
an
challmeondgeel 189
9.S7 ary 199
umm
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