Review of the Food-borne Zoonoses Research ... - ARCHIVE: Defra

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Review summary A key aim of this project was to express recombinant antibodies in edible crop plants, to facilitate the delivery of passive immunisation against Escherichia coli O157:H7 to mucosal surfaces of livestock. Recombinant antibodies were produced, however these were less protective against E. coli O157 adherence factors than traditionally produced antibodies. As a consequence, the assays with transgenic plants were not performed. This was an ambitious project which was more in the area of pure research rather than applied research, however, it fitted well with the Defra request at the time for novel control methodologies. The work was useful preliminary research to determine whether immunisation might be an option. 88
Project code: OZ 0704 Project title: Quantification of E. coli O157:H7 virulence factors in vivo using real-time RT-PCR Start date (dd/mm/yy): 01/06/1999 End date (dd/mm/yy): 30/09/2002 £200,764 Total cost: Affiliation: Dept. Vet. Pathology, University of Glasgow Sub-contractor(s): None Abstract of research The asymtomatic carrier bovine is a major reservoir of the food borne zoonotic E.coli O157:H7. Previous studies have shown that an individual animal may shed organisms intermittently throughout life and that infection/colonisation can be cleared but that the same animal can also become reinfected. The main site of carriage in the adult bovine is the colon although organisms have been described colonising other regions of the alimentary tract in the adult carrier animal. E.coli O157:H7 has been reported to cause disease in young calves and we have recently identified O157 organisms attached to intestinal epithelium in an adult cow with profuse bloody diarrhoea. Various virulence determinants - VT1, VT2, intimin, fimbrae - have been identified by culturing E.coli O157 in vitro and in studies using mutated organisms. However, whilst the role of some of these determinants in pathogenicity has been established, the precise molecular events associated with colonisation, persistence of infection/carriage and clearance of carriage have not been determined. In this project we will apply the new and exciting technology of real-time, convential PCR and RT-PCR to quantitate the number of organisms and the expression of mRNAs for various virulence factor produced by E.coli O157:H7 organisms in vivo. This will be combined with in situ RT-PCR techniques to obtain a detailed understanding of microbial gene expression during colonisation/ attachment/carriage in the bovine alimentary tract. Thus we will use modern molecular techniques to obtain a clearer understanding of the distribution of infection and pathogenesis (expression of virulence determinants in vivo) of E.coli O157:H7 in cattle. These data would underpin future studies investigating in detail host responses in the carrier animal, relating these to microbial virulence factors and host-pathogen interactions. The overall aim of this approach is to devise strategies leading to reduction/elimination of the E.coli O157:H7 carrier state in cattle. These studies therefore address MAFF policy relating to MAFF Strategic Research Requirements to obtain a clearer understanding of the distrubution of infection, pathology, colonisation, host specificity and virulence of the E.coli O157:H7 bacterium in animals. Such knowledge is essential to the development of preventative and control measures for O157 infection of animals, thereby reducing food contamination and the incidence of O157 food poisoning in man. Development of these techniques for E.coli O157:H7 could also form the basis of investigating host-pathogen interactions of other food borne zoonoses at the molecular level. 89
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Contents Section A - Introduction..
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Review of the Food-borne Zoonoses (
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Plans for taking forward these prio
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Policy Objectives The key policy ob
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Figure 2. Laboratory reports of Sal
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Figure 4. Laboratory reports of Cam
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6. Aims of the FBZ research review
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Section B - Review Summaries 16
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epidemiology of Salmonella and anti
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There is a need to objectively asse
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2. Campylobacter review summary 2.1
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for this organism. Research within
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3. E. coli review summary 3.1 Succe
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4. Other zoonotic pathogens of inte
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Annex 1 - Projects Reviewed 30
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OZ0311 Conventional versus capillar
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OZ0402 OZ0404 OZ0405 OZ0407 OZ0604
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OZ0704 OZ0705 OZ0706 OZ0707 Quantif
Page 38 and 39: Annex 2 - Project Abstracts and Pro
Page 40 and 41: To test the suitability of the AFLP
Page 42 and 43: Project code: OZ0324 Project title:
Page 46 and 47: Review Summary The knowledge that t
Page 48 and 49: Review Summary The project was felt
Page 52 and 53: It is clear that the EC legislative
Page 54 and 55: if the vaccines currently available
Page 56 and 57: Four published papers have come out
Page 58 and 59: Review Summary This epidemiological
Page 60 and 61: Project code: OZO316 Project title:
Page 64 and 65: agricultural industry/public health
Page 66 and 67: testing laboratories. In addition,
Page 68 and 69: animal pathogen but is a human path
Page 70 and 71: esearch has also substantially impr
Page 72 and 73: to elucidate the antigenic repertoi
Page 74 and 75: and have contributed to our underst
Page 76 and 77: Fundamental issues concerning the f
Page 78 and 79: Review summary Project OZ0606 focus
Page 80 and 81: Abstract of research OZ0608 The ado
Page 82 and 83: thinning process for molecular trac
Page 84 and 85: Project code: LK0665 Project title:
Page 86 and 87: Review summary While poultry meat i
Page 90 and 91: Review summary This project aimed t
Page 96 and 97: Project code: OZ0138C Project title
Page 102 and 103: Abstract of research Projects OZ070
Page 104 and 105: produced were both relevant and imp
Page 106 and 107: List of External Reviewers. Mr. Pau
Page 108 and 109: Review of Defra’s FBZ Research Pr
Page 110 and 111: Review of Defra’s FBZ Research Pr
Page 112 and 113: Annex 5 - Review Timetable 112
Page 114 and 115: 11.45 - 12.00 Bovine and Porcine Fo
Page 116: Campylobacter Other 11.50 - 12.05 P
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