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

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Project code: OZ0311 Project title: Conventional versus capillary electrophoresis of RFLP fragments and PCR products for sensitive and rapid subtyping of Salmonella. Start date (dd/mm/yy): 01/06/99 End date (dd/mm/yy): 31/03/04 £373,347 Total cost: Affiliation: Central Science Laboratory Abstract of research We compared different molecular typing techniques in combination with two electrophoretic techniques (conventional gel and capillary electrophoresis) for molecular subtyping of a wide range of Salmonella enterica serovars from animal, human and food or feed sources. These included Pulsed-field Gel Electrophoresis (PFGE), Restriction Fragment Length Polymorhism (RFLP) and polymerase chain (PCR) based techniques such as Fluorescent Amplified Restriction Fragment Length Polymorphism (f- AFLP), Random Amplification of Polymorphic DNA (RAPD), Repetitive Extragenic Palindromic (REP) and Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR.Comparisons between gel based and capillary electrophoresis showed that gel electrophoresis was more reliable and gave greater band resolution than capillary electrophoresis based the equipment available. Most methods gave good but not perfect serovar identification. PFGE and f-AFLP were comparable and were the most promising techniques for differentiation and characterisation at strain level within serovars. f-AFLP analysis requires improved standardisation including use of internal standards and triplicate analysis. Review Summary This was an important comparison of techniques aimed at improving typing methods for salmonella, with the approaches used valid at the time of funding. Unfortunately, none of the methods delivered the sensitivity required for epidemiological studies. Some useful information was obtained which informed future approaches to molecular typing. A major criticism of the study was that the work carried had not been published in the scientific literature as the findings would have been useful to other scientists in the field. 44
Project code: OZ0314 Project title: The role of defined bacterial genes and host genetic background in intestinal colonisation of poultry by Salmonella Start date 01/04/99 (dd/mm/yy): End date (dd/mm/yy): 30/06/02 Affiliation: Institute for Animal Health Project code: OZ0320 Project title: Bacterial and host genes in Salmonella colonisation in poultry Start date 01/07/02 (dd/mm/yy): End date 30/06/05 (dd/mm/yy): £956,279 Total cost: Affiliation: Institute for Animal Health Abstract of research We have demonstrated that reproducible differences exist in the extent to which several Salmonella serovars colonise the alimentrary tract of genetically different inbred populations of chickens. This is not associated with the MHC type, is not sex linked and not the result of SAL1 or NRAMP1 haplotypes but is a dominant trait. Similar patterns are found when chickens are also infected with Campylobacter jejuni indicating a common mechanism. Considerable differences in the patterns of expression of selected defensin have been found in the intestinal tissues. No major differences could be found in the extent to which mucin obtained from resistant or susceptible lines of chickens were able to act as nutrient sources for colonising Salmonella serovars. The resistance characteristic has enormous potential for selective breeding and for use with vaccines for infection control. Bacterial mutational studies indicated that ability to utilise sialic acid was not essential to colonisation. A number of electron acceptors used under anaerobic conditions were tested and none was found to be essential. Greater reductions were seen when mutants were unable to use substrate level phosphorylation suggesting that fermentation may be of greatest importance in the large gut. The energy storage compounds glycogen and polyphosphate contributed little to colonisation whereas they and particularly polyphosphate was involved in survival outside the body suggesting a very important role in homeostasis. 45
Page 2 and 3: Contents Section A - Introduction..
Page 4 and 5: Review of the Food-borne Zoonoses (
Page 6 and 7: Plans for taking forward these prio
Page 8 and 9: Policy Objectives The key policy ob
Page 10 and 11: Figure 2. Laboratory reports of Sal
Page 12 and 13: Figure 4. Laboratory reports of Cam
Page 14 and 15: 6. Aims of the FBZ research review
Page 16 and 17: Section B - Review Summaries 16
Page 18 and 19: epidemiology of Salmonella and anti
Page 20 and 21: There is a need to objectively asse
Page 22 and 23: 2. Campylobacter review summary 2.1
Page 24 and 25: for this organism. Research within
Page 26 and 27: 3. E. coli review summary 3.1 Succe
Page 28 and 29: 4. Other zoonotic pathogens of inte
Page 30 and 31: Annex 1 - Projects Reviewed 30
Page 32 and 33: OZ0311 Conventional versus capillar
Page 34 and 35: OZ0402 OZ0404 OZ0405 OZ0407 OZ0604
Page 36 and 37: 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 88 and 89: Review summary A key aim of this pr
Page 90 and 91: Review summary This project aimed t
Page 94 and 95:
Project code: OZ0712 Project title:
Page 96 and 97:
Project code: OZ0138C Project title
Page 98 and 99:
Page 100 and 101:
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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Review of the Food-borne Zoonoses Research ... - ARCHIVE: Defra

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