Microbiologist, September 2007

The magazine of the Society for Applied Microbiology

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September 2007

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Vol 8 No 3

ISSN 1479-2699

Getting vaccines to those that need them most

INSIDE

TB Vaccines — an update ■ Summer Conference 2007 full report ■ Winter Meeting 2008 ■ Careers: Clinical Research ■ In the loop: PECS column ■ Mediawatch: the tale of a microbiologist and the media ■ Caption Competition ■ Statnote 10 ■ Med-Vet-Net — 3rd Annual Scientific Meeting

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September 2007

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Vol 8 No 3

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ISSN 1479-2699

the magazine of the Society for Applied Microbiology

contents members

04 Editorial: infectious disease in the developing world 05 Contact point: full contact information for the Society 06 Benefits: what SfAM can do for you and how to join us 07 Microbreak: caption competition 08 President’s and CEO’s columns 10 Membership Matters 14 In the loop: PECS column 43 Careers: Clinical Research and the Clinical Research Associate 46 MiSAC Competition 2007 50 Students into Work Grant reports 52 President’s Fund Grant articles

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news

18 MediaWatch: the tale of a microbiologist and the media 20 Med-Vet-Net: 3rd Annual Scientific Meeting 22 Bio Focus: Richard Dyer on competition within the biosciences publications

07 29

Microbreak: caption competion

Winter Meeting 2008: book now!

information

15 JournalWatch: Microbial Biotechnology — a new journal 16 Book Reviews

Microbiologist is published quarterly by the Society for Applied Microbiology. ISSN 1479-2699. Registered in the UK as a charity.

features

© Society for Applied Microbiology 2007. Material published in Microbiologist may not be reproduced, stored in a retrieval system, or transmitted in any form without the prior permission of the Society.

32 Getting vaccines to those that need them most 36 Tuberculosis vaccines — an update 40 Revealing all: the microscopic world of compost 41 Statnote 10

Editor: Lucy Harper. lucy@sfam.org.uk Contributions: These are always welcome and should be addressed to the Editor at: lucy@sfam.org.uk Advertising: Lucy Harper Tel: +44 (0)1234 326709. email: lucy@sfam.org.uk

meetings

Design and print: Pollard Creativity. sfam@pollardcreativity.co.uk

23 Summer Conference 2007: Full report 29 Winter Meeting 2008: programme and booking form

Society for Applied Microbiology, Bedford Heights, Brickhill Drive, Bedford MK41 7PH, UK Tel: +44 (0)1234 326661. Fax: +44 (0)1234 326678

commercial

www.sfam.org.uk

54 Advertisements and news from our Corporate members September 2007

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members

Microbiologist is published quarterly by the Society for Applied Microbiology, a registered charity.

ISSN 1479-2699 Copy Dates: Vol 8 No.4 Dec 2007 Friday 5 October 2007

T

he issue of infectious disease in the developing world is one which, if this years’ G8 Summit is anything to go by, is now a priority in the minds of our world leaders.

One such disease which has long been a problem in the developing world, is tuberculosis (TB) for which the WHO Global Tuberculosis Control report 2007 stated that “the global epidemic is on the threshold of decline”. However, TB remains a serious global health issue: in 2005 there were an estimated 8.8 million new TB cases, 7.4 million in Asia and sub-Saharan Africa and a total of 1.6 million people died of TB, including 195,000 patients infected with HIV. It is infection with HIV Lucy Harper talks about infectious which is the main reason for failure to reach targets disease in the developing world in the control of TB in areas where HIV prevalence is high. In addition, TB is the primary cause of death of people diagnosed with HIV/AIDS. The rapidly increasing incidence of HIV in Sub-Saharan Africa and other parts of the world has the potential to increase the number of HIV-related cases of TB. More recently the emergence of multi drug resistant (MDR) and We are always looking extensively drug resistant (XDR) TB adds further complication to this for enthusiastic writers already complex problem. The misuse or mismanagement of anti-TB drugs who wish to contribute can lead to the development of resistant strains of Mycobacterium articles to the magazine tuberculosis. MDR-TB classifies M. tuberculosis resistant to at least on their chosen isoniazid and rifampicin, with XDR-TB bacteria being resistant to any microbiological subject. fluoroquinolone, and at least one of three injectable second-line drugs For further information (capreomycin, kanamycin, and amikacin). please email the editor, But with one of the themes of this years’ G8 summit being “helping Lucy Harper at: Africa to develop” (http://www.g-8.de/Webs/G8/EN/G8Summit/g8lucy@sfam.org.uk summit.html) and leaders pledging $60billion towards fighting infectious diseases in Africa (HIV/AIDS, Malaria and tuberculosis), this makes for encouraging news in the fight against TB. In this issue of Microbiologist, Professor Gordon Dougan of the Wellcome Trust Sanger Institute looks at global vaccination programmes and how a global approach aims to get vaccines to those who need them most (page 32).

We then focus on TB specifically for our second feature article, looking in some detail at the development of new TB vaccines in a report by Peter Andersen of Statens Serum Institute, Denmark (page 36).

I have painful memories of receiving my BCG vaccination against TB whilst at school. This issue of Microbiologist discusses happier times at school with a report from this years’ MISAC Competition. Participating pupils were asked to design a public information leaflet on Salmonella: from farm to fork. Page 46 has a full report and pictures of the very impressive winning entries.

Finally, we have resurrected the ‘Microbreak’ section of the magazine for this issue. A photo which was taken at this years’ Summer Conference, Microbiology of Fresh Produce, Park Plaza Hotel, Cardiff has been used for a caption competition — take a look at page 7 and send your captions to me to be in with a chance of winning an exclusive prize!

editorial contribute

Lucy Harper

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Vol 9 No.1 March 2008 Friday 4 January 2008 Vol 9 No.2 June 2008 Friday 4 April 2008 Vol 9 No.3 Sept 2008 Friday 4 July 2008 Disclaimer: The Society assumes no responsibility for the opinions expressed by contributors. The views expressed by Society officers and staff do not necessarily represent the official position of the Society.

Readers should note that scientific material is not refereed and represents only the views of the authors.

The claims of advertisers cannot be guaranteed.

Subscriptions: A subscription to Microbiologist is included in the annual SfAM membership fee.

For further information about the many benefits of membership please see page 6.

Advertising: Information about advertising in Microbiologist and how to submit advertisements are can be found on the Society website.

Website: our website (www.sfam.org.uk) is a timely source of upto-date information on all Society matters and maintains a comprehensive archive of articles and reports on a variety of microbiological topics.

contact point

executive committee COMMITTEE MEMBERS 2006 - 2008 HON PRESIDENT: Dr Margaret Patterson, Agri-Food and Biosciences Institute, Agricultural, Food and Environmental Science Division, Newforge Lane, Belfast BT9 5PX email: margaret.patterson@afbini.gov.uk HON VICE PRESIDENT: Professor Geoff Hanlon, School of Pharmacy and Biomolecular Sciences, University of Brighton, Moulsecoomb, Brighton BN2 4GJ email: g.w.hanlon@brighton.ac.uk HON GENERAL SECRETARY: Dr Anthony Hilton, School of Life and Health Sciences, Aston University, Birmingham B4 7ET email: a.c.hilton@aston.ac.uk HON MEETINGS SECRETARY: Professor Martin Adams, School of Biomedical & Molecular Sciences, University of Surrey, Guildford,Surrey GU2 7XH email: m.adams@surrey.ac.uk

Society for Applied Microbiology, Bedford Heights, Brickhill Drive, Bedford MK41 7PH, UK tel: +44 (0)1234 326661 fax: +44 (0)1234 326678 email: communications@sfam.org.uk www.sfam.org.uk

HON TREASURER: Dr Valerie Edwards-Jones, Research Development Unit, Manchester Metropolitan University, Lower Chatham St, Manchester M15 5HA email: v.e.jones@mmu.ac.uk HON EDITOR: Journal of Applied Microbiology Professor Arthur Gilmour, Agri-Food and Biosciences Institute, Agricultural, Food and Environmental Science Division, Newforge Lane, Belfast BT9 5PX email: arthur.gilmour@afbini.gov.uk HON EDITOR: Letters in Applied Microbiology Dr Jean-Yves Maillard, Welsh School of Pharmacy, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff CF10 3XF email: maillardj@cardiff.ac.uk ORDINARY COMMITTEE MEMBERS UNTIL JULY 2008 Dr Tony Worthington, Department of Pharmaceutical and Biological Sciences, Aston University, Birmingham B4 7ET email: T.Worthington@aston.ac.uk Dr Andrew Sails, Health Protection Agency, Institute of Pathology, Newcastle General Hospital, Westgate Road, Newcastle-upon-Tyne NE4 6BE email: andrew.sails@hpa.org.uk ORDINARY COMMITTEE MEMBERS UNTIL JULY 2009 Professor Carol Phillips, School of Health, The University of Northampton, Boughton Green Road, Northampton NN2 7AL email: Carol.Phillips@northampton.ac.uk Dr Mark Fielder, School of Life Sciences, Kingston University, Penrhyn Road, Kingston upon Thames, Surrey KT1 2EE email: m.fielder@kingston.ac.uk Professor Joanna Verran, Manchester Metropolitan University, Dept Biological Sciences, Chester Street, Manchester M1 5GD email: j.verran@mmu.ac.uk

society office staff CHIEF EXECUTIVE OFFICER: Philip Wheat email: pfwheat@sfam.org.uk tel: 01234 326661

ORDINARY COMMITTEE MEMBERS UNTIL JULY 2010 Mr Steve Davies MA CSci FIBMS, Microbiology Department, Northern General Hospital, Herries Road, Sheffield, S7 5AU email: steve.davies@sth.nhs.uk

COMMUNICATIONS OFFICER: Lucy Harper email: lucy@sfam.org.uk tel: 01234 326709

Dr Louise Fielding, Food Research and Consultancy Unit, Cardiff School of Health Sciences, University of Wales Institute Cardiff, Llandaff Campus, Western Avenue, Cardiff, CF5 2YB email: lfielding@uwic.ac.uk

MEMBERSHIP CO-ORDINATOR: Julie Wright email: julie@sfam.org.uk tel: 01234 326846

Professor Andrew Fox, Health Protection Agency North West, PO Box 209, Clinical Sciences Building, Manchester Royal Infirmary, Manchester, M13 9WZ email:andrew.fox@hpa.org.uk

ADMINISTRATOR: Ruth Sawford email: ruth@sfam.org.uk tel: 01234 326661

Dr. Andrew McBain, School of Pharmacy & Pharmaceutical Sciences, Stopford Building, University of Manchester, Manchester, M13 9PT email: andrew.mcbain@manchester.ac.uk

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membership benefits options ■ Full ordinary membership gives online access to the Journal of Applied Microbiology, Letters in Applied Microbiology and Environmental Microbiology, copies of Microbiologist, preferential registration rates at Society meetings and access to the members areas of the website.

The Society for Applied Microbiology is the voice of applied microbiology within the UK and was founded in 1931. Society members play a leading role in shaping the future of applied microbiology, and enjoy many benefits, including: ■ Substantially reduced rates for attendance at Society meetings and conferences ■ Access to the members areas of the Society website ■ Many generous grants and awards ■ FREE access to three acclaimed journals Detailed information about all these benefits and more can be found on the Society website at: www.sfam.org.uk GRANTS & AWARDS: Many grants, awards and prizes are available to members including the W H Pierce Memorial Prize and Prizes for Student Oral Presentations and Posters at the Summer Conference. In addition to these substantial awards, the Society has funds to assist members in their careers as microbiologists. These include The President’s Fund, Conference Studentships, Sponsored Lectures and the popular Students into Work Scheme.

Full details of all the Society’s grants and awards can be found on the website together with PDF application forms available to download from the members area.

JOURNALS: The Society publishes two monthly journals: Journal of Applied Microbiology and Letters in Applied Microbiology. We also produce this quarterly colour magazine, Microbiologist, which contains features, topical news stories and full details of our meetings. The Society is also a partner with Blackwell Publishing in the monthly journal Environmental Microbiology and we are launching a new journal for 2008; Microbial Biotechnology.

Synergy is an online service provided by Blackwell Publishing that gives Full and Student Members FREE access to the online versions of the Society’s three journals: Journal of Applied Microbiology, Letters in Applied Microbiology, Environmental Microbiology and Microbial Biotechnology. Members can register for this service at http://www.blackwell-science.com. Members can also submit papers directly to our journals via an online submission service. For more information about Synergy or online manuscript submission, please visit the website.

MEETINGS: We hold two annual meetings. The January Meeting is a one-day meeting with parallel sessions on topical subjects. The Summer Conference is held every July and comprises a main symposium, a poster session, the AGM and a lively social programme. We also hold occasional joint ventures with other organisations on topics of mutual interest.

WEBSITE: The website is the best source of detailed information on the Society and it’s many activities. It has fully interactive membership areas where you can book your place at Society meetings, find exclusive SfAM documentation and much more.

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■ Full student membership confers the same benefits as Full Membership at a specially reduced rate for full time students not in receipt of a taxable salary.

■ Associate membership is only open to those with an interest in applied microbiology without it being a prime aspect of their job. For example, school teachers and those taking a career break; on maternity leave, or working temporarily in other areas. It does not provide access to any journals or Society grants and awards.

■ Honorary membership of the Society is by election only and this honour is conferred on persons of distinction in the field of applied microbiology. Honorary members have access to our online journals.

■ Corporate membership is open to all companies with an interest in microbiology.

Corporate members benefits include: ● Quarter page advertisement in each issue of Microbiologist (which can be upgraded to a larger size at discounted rates) ● the opportunity to publish press releases, company news, etc., in each issue of Microbiologist ● Full page advertisement in the Members' Handbook. ● FREE banner advert on the Society Website with a direct link to your company site. ● Up to three members of company staff attending Society meetings at members' rate (This means a 50% discount on non member registration rate).

■ Retirement membership is available to Full Members once they have retired from their employment. Retired members are entitled to all the benefits of Full Membership except grants and access to Journal of Applied Microbiology, Letters in Applied Microbiology and Environmental Microbiology.

JOIN US! You can apply for membership on, or offline. To apply offline, please contact the Membership Coordinator, Julie Wright on +44 (0)1234 326846, or email julie@sfam.org.uk. Alternatively, write to her at: The Society for Applied Microbiology, Bedford Heights, Brickhill Drive, Bedford MK41 7PH, UK

www.sfam.org.uk

microbreak This picture was taken during the tour of Cardiff’s Millennium Stadium prior to this years’ SfAM Summer Conference dinner. We thought it was too good a photograph to leave to the archive and we’re looking for a witty caption.

If you can think of a suitably humorous comment (keep it clean please), send it by email to the Editor no later than 12 October 2007 to be in with a chance of winning a prize.

You know you’re a microbiologist when... ✔ The phrase ‘fermentation seminar’ is slang for having a beer.

✔ You bet on said disease outbreaks. ✔ You think that the inventor of parafilm was more of a genius than Einstein.

✔ Family and friends avoid asking how work/school was today during dinner. ✔ You yell at the doctor for prescribing antibiotics to you after diagnosing you with a cold. ✔ Friends come into the lab and retch from the stench, but you don’t notice it.

✔ You’ve ever streaked a plate of Streptococci with your initials. ✔ You remember Latin species names easier than your friends’ names. ✔ When flaming your loop you’ve thought that you are committing mass genocide—and giggled!

✔ You talk about Koch’s postulates and wonder why people giggle. ✔ You’ve seriously considered a biohazard tattoo.

✔ You can roughly guess the bacteria by the colony morphology.

✔ You’ve wondered what LB broth tastes like.

✔ You know what ‘colony morphology’ means.

✔ You follow disease outbreaks with more enthusiasm than your favourite sport.

✔ You can self-diagnose food poisoning due to the amount of time between eating and symptoms.

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I

would like to thank all who attended the summer conference in Cardiff in July — Martin Adams and his team for organising the programme, the speakers, and all the attendees.

The title, ‘Microbiology of Fresh Produce’, was extremely topical and we had over 150 delegates from 13 different countries around the world.

You will see from the conference report (page 23) that, not only was the science excellent but there were lots of opportunities to interact with other microbiologists during the packed social programme.

Many thanks also to those delegates who returned the conference feedback forms. We are always keen to receive feedback from our members at any time — good or bad. We endeavour to ensure the Society meets the needs and expectations of its Your views are important to us members so any suggestions or ideas will Margaret Patterson invites all members to be considered seriously participate in the many new developments

by Committee and the Society is considering by providing her wherever possible, they will be taken forward. A with your views and suggestions good example of this is the formation of the Postgraduate and Early Career Scientists subcommittee (PECS). We received some feedback from student members that they would like to be better identified within SfAM. As a result, Dr Anthony Hilton has been working with all interested postgraduate student members to develop programmes aimed specifically at this group. The remit of the group has since evolved to include those who have just started their scientific career — and so PECS was born (see page 14 for their regular column).

Another example of responding to feedback has been the one-day Spring meeting aimed at Biomedical Scientists. The first meeting was held last April in Manchester, and was extremely popular. As a result we are in the process of planning a similar event again next year.

The Society is keen to embrace changes in new technologies wherever appropriate. The website continues to develop and improve and we will soon provide the facility for nonmembers to apply on-line for membership and soon delegates will also be able to register for conferences online. This is something that many members have requested and will ease the application process. The ‘microbiology in the news’ section is updated regularly, and we hope this is a useful resource. We are constantly on the lookout for new ideas and feedback for the website so all suggestions are appreciated.

We have recently updated the membership database allowing us to send email bulletins to

president’s column

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members every month. We can also target specific groups of our membership based on your interests so that we alert you to activities relevant to you. We feel this alerts our members to all relevant issues in a timely way but would welcome your views on its usefulness or suggestions as to how this system could be improved. There are many other ways the Society can use new technologies to communicate with members, and the general public about applied microbiology in general.

For example, potentially we could make keynote lectures at our conferences available as podcasts — do you think this would be worthwhile? Or perhaps you would like to see a wider SfAM presence online or you think podcasts would be put to better use as a commentary on recent Microbiology news events?

The Committee are considering these and many other ideas for improving benefits to our members, and informing the community in general about applied microbiology. If you have any comments on current activities or suggestions for the future, please do not hesitate contact us. You can write to me directly (contact details on page 5) or the Society Office. We are especially aware that it is the members that form the backbone of SfAM and we are keen to hear from you all.

Dr Margaret Patterson President of the Society

29 - 31). The meeting following this will be the second in the series ‘Broadening Microbiology Horizons’ which will be primarily targeting personnel who work in Clinical Microbiology laboratories. This one day meeting will take place on the 9 April 2008 at the excellent Lakeside Conference facilities, Aston University, Birmingham. Finally, the 2008 summer conference will be held between 7 – 10 July at the Wellington Park Hotel, Belfast where the topic covered will be Water Microbiology. Full details of all the meetings will appear shortly on the Society’s web site and in future issues of Microbiologist.

Attendance at Society’s meetings is increasing so I strongly recommend you book your place early to avoid disappointment.

During the second week of July we attended Philip Wheat reports on the latest the International Association of Food developments within the Society Protection (IAFP) meeting in Orlando, USA.

ceo’s column

I

am writing this column the first few weeks of July. The month so far has been an extremely busy but rewarding one.

July started with a very successful summer conference in Cardiff. Over one hundred and fifty people attended over the four-day event.

The conference was so popular that we declared the meeting full which unfortunately resulted in us having to turn away delegates who tried to book just before the event. It was a great pleasure meeting all the delegates who attended.

In particular, it was very pleasing to see that the conference attracted delegates from over 13 countries from around the world. This probably reflects the very topical (Microbiology of Fresh Produce) nature of the meeting after recent food scares with spinach and other fresh produce. The programme was very well received by all delegates. Professor Martin Adams and the rest of the Meetings subcommittee are all to be congratulated in producing such a comprehensive and informative programme covering the area. I look forward during 2008 to seeing many of the papers from the conference published in the Journal of Applied Microbiology. In addition to the scientific programme, delegates also enjoyed an extensive array of social events (see page 23 for a full conference report).

Plans are already in place for the calendar of 2008 meetings. These begin at the Royal Society in London on the 9 January. This one day winter meeting will cover both quality control and accreditation in applied microbiology and the microbiology of alcoholic beverages (see pages

During this three day conference it was a great pleasure to meet many existing members who stopped at the Society’s exhibition stand. In addition, several new members were enlisted and many other potential members requested further information about joining the Society. These included several potential new corporate members. In addition to the new members and potential new members we also enhanced our relationship with the Executive Committee members of the IAFP. Finally, I am delighted to inform you that the office has a new member of staff. Ruth Sawford joined the office staff in July. Ruth works as an Office Administrator and has much experience in administration duties (see page 10 for a full introduction).

Philp Wheat Chief Executive Officer

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Membership Changes New Members We would like to warmly welcome the following new members and hope that you will participate fully in the activities of the Society.

Australia S. Gaskin

Austria A. Sessitsch

Belgium I. Habib; K. Houf

Denmark

Welcome!

We extend a warm welcome to Ruth Sawford, newly appointed Administrator for SfAM. I started as Administrator with SfAM in early July and am finding it is very different from my previous roles in the engineering sector, in both size, subject matter and in the companies and individuals I have dealings with.

My previous role was in a large, worldwide converting equipment company where I was first Secretary/PA and then took on the role of Credit Controller and administrator, handling many hundreds of accounts covering all time zones, countries and all with very different views on debts and payments which made for interesting correspondence.

I was born in Lincolnshire and moved south many years ago, married a Bedfordshire boy and have two grown up children (David and Jenny).

I enjoy gardening, reading and theatre going and especially enjoy all things historic (houses, plays, books) which I went back to studying as a mature student plus dabbling in art from time to time.

Hopefully my background in administration, attention to detail and ability to multi-task will provide a good basis for assisting the Society to run in a smooth and efficient manner.

membership matters

Ruth Sawford

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E. Litrup

France C. Moran; V. Thomas

Ireland K. Considine; B. V. Jones; G. Moschonos; C. O’Reilly; S. Patil

Kuwait E. Alsaif

Mexico S. R. Treso-Estrada

Nigeria J. C. Orji

United Kingdom B. A. F. Blair; M. Al-Sallaqi; Al-Zeyara; A. Anyogu; B. Awamaria; B. Banja; P. Chanos; M.A. Chattaway; P. Danes; S. B. Hanniffy; A. Hashim; S. Kaur; R. A. J. Nicholas; E. O’Gara; K. O’Malley; N. S. ; T. K. RalebitsoSenior; C. Rice; N. Sharef A Ben Muhamed; R. G. Stentz; E. Szwed; A. Vanichpun; L. Vasireddy; P. L. Waines; J. M. Ward; R. L. Ward; J. S. Webb

USA Y. Ahmed; S. Bornstein-Forst; J. Byron; G. Chapman; J. LeJeune; K. R. Min; M. W. Shepherd; G. Siragusa; J. E. M. Watts

CORPORATE Dow Chemical Company USA

LOSSES The Society was saddened to learn of the death of the following member: C. M. Cousins, Retired Member from St Ives, Cornwall, who sadly died on 28 June 2007.

2007 SfAM AGM The 76th annual general meeting of the Society for Applied Microbiology was held on Wednesday 4 July at 4.30 pm at the Park Plaza Hotel, Cardiff. The Honorary President, Dr Margaret Patterson, was in the chair attending the meeting. This was accepted.

Present Margaret Patterson, Anthony Hilton, Karen Stanley, Christine Dodd, Will Waites, Martin Adams, Sally Cutler, Mark Fielder, Tony Worthington, Andrew Sails, Louise Fielding, Don Whitley, Susannah Walsh, Carol Phillips, Peter Green, Valerie Edwards-Jones, Bernard Mackey, Janet Corry, Andrew Hall, Lucy Harper, Adrian Peters, Geoff Hanlon, Gillian Francis, Bernard Dixon, Maurice Moss, Catherine Ramsay, R. W. A. Park, Jeffrey McGarvey, Bill Keevil.

1. Apologies for absence Apologies were received from Peter Silley, Sue Passmore, Arthur Gilmour, Muriel Rhodes-Roberts, David Post and John Rigarlsford.

2. 75th Annual Meeting The minutes of the 75th Annual Meeting held in Edinburgh, 2006, were approved and accepted as correct by those present.

Proposed: Mark Fielder Seconded: Alan Gardner

3. Matters arising Philip Wheat provided an update on progress towards Incorporation. Trustees are working on the articles and memorandum of association. Is it anticipated that within eight weeks the governing documents will be lodged with the Charities Commission and Companies House.

5. Adoption of the Annual Report 2006 Dr Patterson asked for the report to be officially adopted by those present.

Proposed: Susannah Walsh.

Seconded: Robert Park.

8. Election of new members

6. Election of the Honorary Vice President Dr Margaret Patterson explained that she will be retiring from her post as President of the Society at the next AGM in July 2008 and the formal written procedure was followed in finding a replacement for this post. A panel comprising three members of Main Committee met to discuss potential candidates and four candidates were selected. Members of the Committee then took a vote and Professor Geoff Hanlon was duly nominated. Margaret Patterson then asked for any further nominations from the floor. No further nominations were made.

Dr Patterson congratulated Professor Hanlon. Seconded: Sally Cutler.

7. Election of new Committee members

Report of the Society for the year 2006.

Copies of the report for the year 2006 were previously distributed to all members

Dr Patterson then stated that four nominations had been made to committee.

The new members of committee for 2007-

Photo Competition

4. Report of the Trustees

Dr Margaret Patterson reported that this year there were four committee vacancies as Dr Susannah Walsh, Dr John Coote, Professor Geoff Hanlon and Dr Karen Stanley were retiring by rotation. Dr Hilton thanked these four people for their contributions and hard work during their terms in office.

Have you taken an outstanding photograph of your beloved bugs? Do you know someone who has and you'd like to see their work in print?

Perhaps you've taken a photograph while attending a SfAM conference, which you think is worthy of reproduction?

Due to popular demand, SfAM are running the photography competition again this year. We are looking for twelve eye-catching images to use for our 2008 calendar which we will be giving to all our members as a Christmas gift.

To enter this competition, please send your photographs to the Editor in the form of JPEG files which must be a minimum size of 7 x 7cm at

2010 are: Professor Andrew Fox, Dr Louise Fielding, Dr Andrew McBain and Mr Steve Davies.

Dr Patterson welcomed Dr Louise Fielding — the only new committee member present.

This was proposed by Mark Fielder and seconded by Peter Green.

Dr Anthony Hilton provided a list of names of applicants for ordinary membership which was tabled. During 2006 there are 1,207 true members of the Society showing an increase in membership.

This has been published in the Microbiologist throughout the year.

Dr Hilton holds a summary of the deaths and resignations of members through the previous year for consultation if requested.

9. Any other business Will Waites suggested that the conference badges and delegate list provided for the summer conference were variable. He explained that the affiliation of some delegates was not very clear. Philip Wheat suggested that a space be added to the booking forms allowing delegates to choose the affiliation information placed on badges.

Maurice Moss then thanked the Society for all the work that had gone into the organisation of a very successful summer conference.

10. Date of next meeting The next ordinary AGM will be on 9th July 2008 in Belfast.

300dpi (800 x 800 pixels). Alternatively, you can send the original photographs in hard copy to the Society Office and we will return them to you once they have been scanned.

Photographs will appear in one of two categories: 1. Scientific — e.g., a colourful image using bacteria.

2. Non-scientific but with a SfAM theme e.g., taken at a SfAM event.

The closing date for entries for this competition is 28 September 2007.

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In Memoriam Professor Malcolm Woodbine, Emeritus Professor of Agricultural Microbiology, University of Nottingham died on 7 February 2007, aged 88.

Malcolm was educated at Lancaster Royal Grammar school. He left school at 16 and later won a Lancashire County Council Scholarship to read Chemistry at the University of Manchester.

He graduated in 1941 and as a Quaker and registered conscientious objector, joined Wellcome Research Laboratories to work on developing the early antibiotics. He gained his MSc in 1948 and became a Scientific Advisor in the Scientific Civil Service, Ministry of Food. He was seconded to the Medical Research Council to carry out research on the potential production of fats from moulds under the direction of Professor T.K. Walker at University of Manchester Institute of Science and Technology (UMIST) and gained a PhD for this work. He returned to the Ministry of Food to work on the accelerated freeze-drying of foods.

In 1952, he was appointed to one of the first Senior Lectureships in the University of Nottingham at the School of Agriculture. He was promoted to a Readership in 1959 and was invited to spend a year in the USA on a Senior Foreign Scientist Fellowship in 1969. He was appointed to a personal chair in 1976, retiring in 1983 at the age of 65.

His research interests embraced the conservation and utilisation of resources from which came three international symposia on the prudent use of antibiotics (1964, 1976, and 1983). Much of his life’s work was directed to Listeria monocytogenes.

Cathy Woodbine (daughter)

Dr Timothy N. Whitmore After postdoctoral research at the University of Warwick, Tim joined the staff of the Water Research Centre plc (WRc) at Medmenham in 1989. His work there was particularly associated with the recovery and detection of Cryptosporidium oocysts from potable water and the environment, and with investigating methods to remove oocysts during water treatment. After leaving WRc in 2003, Tim continued related work in the commercial sector. On the Editorial Board of the Journal of Applied Microbiology and Letters in Applied Microbiology for many years, Tim was a very versatile and enthusiastic applied microbiologist, and his quiet humour will be sorely missed by his family, friends and colleagues.

Stefan Sidorowicz

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mailbox From: Professor Jim Lynch, OBE Subject: OECD, OBE and Microbiology I was of course surprised and delighted to receive the OBE in the Queen’s Birthday Honours for my work over the past 17 years as coordinator of the OECD Co-operative Research Programme on Biological Resource Management for Sustainable Agricultural Systems. Applied microbiology fits very firmly into the Organisation for Economic Cooperation and Development (OECD) Programme as a core integrating discipline. We award about 80 bi-national postdoctoral fellowships for periods up to six months each year between the 30 member countries and also sponsor about ten workshops (up to about 30 participants). Theme 1 is The Natural Resources Challenge, Theme 2 is Sustainability in Practice and Theme 3 is The Food Chain. All the information can be found at www.oecd.org/agr/ prog. I have genuinely found that a microbiological background has been very useful in communicating across the wide range of disciplines and environments that are covered by the programme. Increasingly we have interacted with social scientists who have brought new perspectives to what we are doing and not only improving knowledge transfer but also by bringing a sociological, economic and political perspective to the research activity. To my mind, this is what makes applied science exciting.

SfAM would like to congratulate Professor Lynch on his award of OBE at the Queen’s Birthday Honours.

From: Eshwar Mahenthiralingam Subject: Students into Work success Ceri Wilmot, a second year microbiology student at Cardiff University, started a Students into Work project with me in July, and is searching for B. cepacia complex isolates that produce antimicrobials that can inhibit other B. cepacia bacteria as well as cystic fibrosis (CF) pathogens such as Staphylococcus aureus and Pseudomonas aeruginosa. At the time of going to press and after screening only a couple of dozen isolates, the project has already been successful in identifying an environmental B. cepacia complex strain which produces a potent inhibitor of Burkholderia multivorans, the second most dominant CF pathogen in the complex. Hopefully, after the 10 week project we will know a lot more information about antibacterial agents produced by B. cepacia complex bacteria and whether these are worthy of further development as clinically useful antibiotics. Ceri is really enjoying the project and finding it is providing her with invaluable laboratory experience. We’d like to thank SfAM for awarding her this grant which has been of great value to both of us.

2007 W H Pierce prize winner: Dr Dennis Linton

Dr Dennis Linton (left) receiving his prize

On 4 July 2007, Colin Booth of Oxoid presented the W H Pierce prize for young microbiologists who have made a substantial contribution to the science, to Dr Dennis Linton of the University of Manchester. Here he introduces himself and gives a brief overview of his career.

Following graduation from the University of East Anglia with a degree in Microbiology, I worked as a Clinical Scientist at the Central Public Health Laboratory (now the Centre for Infections) in London. It was there, whilst working for Dr Robert J. Owen and subsequently Dr John Stanley, that I was introduced to the Campylobacter and Helicobacter species that remain my research focus. I was initially involved in developing molecular typing methods for the gastric pathogen Helicobacter pylori and then, as part of my PhD work, in characterising novel Campylobacter and Helicobacter species that had been isolated by Dr Andre Burnens, at the University of Berne.

On completion of my PhD in 1997, I left public health microbiology for academia joining Professor Brendan Wren’s laboratory, then based at St. Bartholomew’s hospital in London. This was a joint project, with Dr Norman Gregson from Guy’s Hospital, investigating the link between Campylobacter jejuni infection in humans and the subsequent development of Guillain-Barre syndrome (GBS), an acute paralytic neuropathy. It is thought that the production of antibodies against the surface located C. jejuni lipooligosaccharide (LOS) results in development of GBS due to the structural similarity between LOS and gangliosides present in human nerve.

Autoimmune-mediated nerve damage can follow leading to paralysis and even death. I was able to identify a number of C. jejuni enzymes that were involved in biosynthesis of LOS molecules

structurally related to gangliosides. It was while studying these genes that I became interested in the biosynthesis of other components of the C. jejuni cell surface including capsular polysaccharide and glycoproteins, as well as the role of phase variable genes in modifying these structures.

During my second postdoctoral position in Professor Wren’s laboratory, now located at the London School of Hygiene and Tropical Medicine, I developed methods for the purification of an intriguing set of previously uncharacterised, surface-located C. jejuni glycoproteins.

Collaboration with Dr Michael Wacker from Professor Markus Aebi’s group in Zurich, led to the identification of a complete C. jejuni N-linked protein glycosylation system, the first such system identified in a bacterial species.

In 2003 I took up a four year Wellcome Trust Career Development Fellowship at the University of Manchester to study the role of C. jejuni phase variable genes in generating cell surface structural diversity. Currently, as a Lecturer I combine research work with the challenges of undergraduate teaching and obtaining funding for my nascent laboratory.

Research is a collaborative venture and I would like to acknowledge the contributions of Professor Markus Aebi, Professor Anne Dell, Dr Paul Hitchen and Dr Andrew Lawson. Furthermore, I am fortunate to have been mentored by a number of outstanding microbiologists including Dr John Stanley, Dr Andrey Karlyshev, Professor Brendan Wren and Professor Ian Roberts. Finally I gratefully acknowledge the financial support of the Wellcome Trust and BBSRC. It is a great honour to be awarded the 2007 W H Pierce prize and I thank both the Society for Applied Microbiology and the sponsors Oxoid.

Sponsor a new Member of the Society and win a £50 Voucher of your choice!

If you feel you could be our next winner for 2007, and would like some promotional material to help you recruit new members please contact Julie Wright, Membership Co-ordinator on 01234 326661 or email julie@sfam.org.uk.

September 2007

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members

I love it — you never get the opportunity to do anything like this at school!

Jo Heaton, Chair of the PECS subcommittee discusses a microbiology workshop she ran for gifted and talented children During August of 2006 I was lucky enough to teach 19 gifted and talented children, attending a two-week National Academy for Gifted and Talented Youth (NAGTY) summer school. I prepared a course entitled ‘Disease Detectives’, which aimed to introduce the students to microbiology and molecular biology, bringing the two subjects together by employing molecular techniques to investigate a mock food-poisoning outbreak. The course was a great success and working with children who were seeing the microbial world for the first time rejuvenated my love for the subject.

in the loop News from the SfAM Post-Graduate and Early-Career Scientist Committee We started ‘large’, examining different species of protozoa and determining the mechanisms used in feeding. We also examined the feeding of the metazoan Daphnia magna and a number of Rotifera, which many of the students wanted to keep as pets (I’m fairly certain some were smuggled home!). Once the basics of using a light microscope were mastered, we moved onto bench skills, practicing Gram stains on probiotic yoghurt drinks and discussing the likelihood of such products being effective. The highlight of the week was our tour around a sewage treatment works in Warrington.

The process engineer described every aspect in detail but the class questioned him tirelessly. We had to drag the boys away from the Archimedes screw pump, which seemed to fascinate them! Once back in the lab we compared the water quality of the final effluent to bathing water samples from Morecambe bay and water from the River Lune. The claim that the effluent was clean 14

enough to drink was soon quashed as the class judged the microbial quality to be “ugh…disgusting”.

Virulence factors and pathogenicity were next on the agenda. The class took the complex ideas in their stride and designed their own ‘bacterium’, explaining the virulence factors it possessed, how it avoided host defences and the symptoms it caused.

All kinds of weird and wonderful infections were proposed, including a neurological disorder which caused the host to sing continuously and a systematic infection resulting in tissue melting.

We then moved on to talking about infection. The class were given bacterial isolates from a group of fictional patients and case notes. I gave a quick lecture introducing them to the oxidase, catalase and coagulase tests and then they were left to determine the bacteria responsible for each infection, using an identification flow chart and additional information on each of the ‘usual suspects’ (Staphylococcus, Streptococcus, E. coli, Bacillus cereus and Pseudomonas aeruginosa). The students correctly identified the infectious agents and went on to research and recommend the necessary treatment.

The group were now confident in their microbiological skills and so we set up a food poisoning scenario. The first task was to use chromogenic and selective agars to isolate E. coli from a range of suspected food items. The chocolate éclair was found to be responsible and the bacterium was identified using API 20E kits, kindly supplied free of charge by Biomeriuex.

Week two concentrated on molecular biology and was co-ordinated by my colleague, Dr Christine Shirras. Each morning session consisted of lectures on DNA structure, inheritance and genetic manipulation. In the lab, the students were introduced to the technique of gel electrophoresis. We returned to the food poisoning outbreak, using PFGE to determine if further E. coli infections were related to the contaminated chocolate éclair.

Several other cases were discovered and an outbreak report was prepared.

On Wednesday morning the students entered the lab to find a crime had been committed! Police tape marked off a blood splattered area, with a broken conical flask, an overturned trolley and

September 2007

a chalk outline: our lab technician had been murdered! The class got on the case immediately, swabbing the blood samples, taking notes and analysing the crime scene. The conclusion was that the victim had been attacked from behind with a stool and that the blood leading away from the area belonged to the murderer. The whole class (and the teachers) were suspects and had to be included in the inquiry. Everybody isolated their own DNA and carried out pulse field gel electrophoresis (PFGE) analysis to create DNA fingerprints.

Comparison of our unique fingerprints to the blood sample suggested that I was the murderer, but I protested my innocence! The reliability of such evidence was debated and the class decided that extra evidence was necessary for a conviction — luckily for me!

On the final day, the students from all the other subject areas gave presentations on their experiences and we held a scientific poster session during lunch. This was a great opportunity for the group to really show-off everything that they had learnt, describing their new skills in detail and using plenty of complex terminology. We received extremely favourable feedback from the students and their parents alike and as a result are running the same course this August. I can’t wait to be inspired again by the students’ enthusiasm and relentless questioning- I’m sure many of them will be fantastic scientists in the future.

N.B. Don’t worry, we didn’t really murder anyone and all bacterial isolates were classed as category 1 and safe for use.

NAGTY is a government initiative to improve provision for schoolchildren in the top five cent of the ability range.

Jo Heaton Lancaster University

publications

Top 5 most downloaded articles published in Journal of Applied Microbiology in 2007: 1) Microbial biofilms in the human gastrointestinal tract. S. Macfarlane, J.F. Dillon.

Vol. 102, No. 5, May 2007 2) Intestinal bacteria and ageing. E.J. Woodmansey. Vol. 102, No. 5, May 2007 3) Understanding the effects of diet on bacterial metabolism in the large intestine. P. Louis, K.P. Scott, S.H. Duncan, H.J. Flint. Vol. 102, No. 5, May 2007 4) The microbiological quality of hot waterwashed broccoli florets and cut green beans. S.C. Stringer, J. Plowman, M.W. Peck. Vol. 102, No. 1, January 2007

journalWatch News about the Society’s

5) Internal amplification controls have not been journals employed in fungal PCR hence potential false negative results.

R.R.M. Paterson. Vol. 102, No. 1, January 2007 Top 5 most downloaded articles published in Letters in Applied Microbiology in 2007: 1) Screening and mutagenesis of a novel Bacillus pumilus strain producing alkaline protease for dehairing. H.Y. Wang, D.M. Liu, Y. Liu, C.F. Cheng, Q.Y. Ma, Q. Huang, Y.Z. Zhang. Vol. 44, No. 1, January 2007 2) Antifungal activity of thyme (Thymus vulgaris L.) essential oil and thymol against moulds from damp dwellings. M. Segvic Klaric, I. Kosalec, J. Mastelic, E. Pieckova, S. Pepeljnak. Vol. 44, No. 1, January 2007 3) Vanillin production from simple phenols by wine-associated lactic acid bacteria. A. Bloem, A. Bertrand, A. Lonvaud-Funel, G. de Revel. Vol. 44, No. 1, January 2007 4) Evaluation of two viral extraction methods for the detection of human noroviruses in shellfish with conventional and real-time reverse transcriptase PCR. L. Baert, M. Uyttendaele, J. Debevere. Vol. 44, No. 1, January 2007 5) Cultivable bacterial diversity from the human colon. S.H. Duncan, P. Louis, H.J. Flint. Vol. 44, No. 4, April 2007 Top 5 most downloaded articles published in Environmental Microbiology in 2007:

3) Environmental predators as models for bacterial pathogenesis. Hubert Hilbi, Stefan S. Weber, Curdin Ragaz, Yves Nyfeler, Simon Urwyler. Vol. 9, No. 3, March 2007 4) Interactions and competition within the microbial community of the human colon: links between diet and health. Harry J. Flint, Sylvia H. Duncan, Karen P. Scott, Petra Louis. Vol. 9, No. 5, May 2007 5) Real-time microbial ecology. Forest Rohwer.

Vol. 9, No. 1, January 2007

microbial biotechnology A new journal for 2008 ■ You are invited to submit your best research papers to Microbial Biotechnology – a new journal for 2008 from Blackwell Publishing and the Society for Applied Microbiology. Microbial Biotechnology has been created to publish papers of original research reporting significant advances (belonging to the upper 25% in the field) in any aspect of microbial applications, including, but not limited to biotechnologies related to: ✔Green chemistry ✔Primary metabolites ✔Food, beverages and supplements ✔Pharmaceuticals ✔Diagnostics ✔Agriculture ✔Bioenergy ✔Biomining, including oil recovery and processing ✔Bioremediation ✔Biopolymers, biomaterials ✔Protein engineering ✔Functional genomics ✔Bionanotechnology ✔Biosurfactants and bioemulsifiers ✔Systems analysis, modelling ✔Biologically-based analytical methods ✔Metabolic engineering ✔Metabolic design ✔Compatible solutes and bioprotectants ✔Process engineering ✔Biosensors, monitoring systems, quantitative microbial risk assessment ■ Microbial Biotechnology has an editorial board composed of the leaders in the biotechnology and associated fields and a core of experienced and dedicated editors. Articles will be published OnlineEarly towards the end of 2007 and the first issue will appear online at the beginning of 2008.

We look forward to publishing your research in Microbial Biotechnology.

1) Theory and the microbial world. Tom Curtis.

Vol. 9, No. 1, January 2007

Kenneth N. Timmis on behalf of the Editors of Microbial Biotechnology email: kti@helmholtz-hzi.de

2) The human microbiome: eliminating the biomedical/environmental dichotomy in microbial ecology. Ruth E. Ley, Rob Knight, Jeffrey I. Gordon. Vol. 9, No. 1, January 2007

■ Submit your manuscript at: http://mc.manuscriptcentral.com/microbio ■ Visit the journal homepage at: www.microbialbiotech.com

September 2007

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publications

reviewers The Society receives several new books every week from publishers around the world and we are always looking for enthusiastic additional reviewers who have an interest in the subjects covered.

There is an up-to-date list of titles available for review at the Society Office.

To make an offer to review any book simply email the Editor of Microbiologist at: lucy@sfam.org.uk.

In return for your efforts you get to keep the book!

Titles for review and book reviews published in Microbiologist will soon be available on the website.

Food spoilage microorganisms Clive de W. Blackburn Woodhead Publishing Limited, Cambridge. ISBN-13:978-1-85573966-6. Price: ÂŁ160.00 Reviewed by: Dr Evdoxios Psomas Food spoilage microorganisms edited by C. de W. Blackburn meets the needs of professionals whose role involves microbial quality control of the food industry as well as microbiologists, non-specialists and students enrolled in Food Microbiology programmes. It serves both audiences very well by: using an abundance of important information to point to the major spoilage microorganisms groups, and; proposing a wide variety of solutions to deal with possible food spoilage problems.

Controlling microbiological spoilage requires knowledge of possible hazards, their likely occurrence in different products, their physiological properties and finally the availability and effectiveness of different preventive measures. All the above important information is covered in detail in this book The book is divided into five parts and each part is subdivided into four or five chapters. Every chapter starts with a concise and helpful summary introducing the reader to the topic discussed in it. Furthermore, chapters are authored by many of the research

book reviews

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September 2007

leaders in Food Microbiology. For example, Dr Kurtzman on detectionidentification-enumeration for spoilage yeasts, Dr Deak on Candida and related genera, Dr Fleet on Saccharomyces, Dr Moss on general characteristics of moulds, Dr Schillinger and Dr Holzapfel on Lactic acid bacteria, and many others.

The first section of the book discusses the detection and analysis of microbial food spoilage. More specifically it is a tightly written review of how to detect and analyse microbial spoilage using the appropriate tools, techniques and methods along with a special reference to predictive modelling. An important chapter of this section, is that on shelf-life which deals with determining the stability and the self life of the foods. I found this chapter extremely interesting and helpful especially for those working for the Food industry.

The second section covers the possible ways of managing microbial food spoilage with particular reference to some of the major food groups where the types of spoilage, the causative microorganisms and methods for control are considered by product type. Included in this section are a well written chapter about microbial food spoilage in cereal and baking products and a chapter about spoilage in the dairy industry. In addition, this section contains a rather descriptive chapter on meat microbial spoilage providing a lot of useful tools on how to cope with this kind of problem.

The following three sections of the book are dedicated to yeasts, moulds, and bacteria, and take a more detailed look at the major organisms responsible for food spoilage. In each chapter the taxonomy, spoilage characteristics, methods for detection and control options are discussed in detail. The control measures proposed by the authors of the above chapters are very helpful.

In summary, I found the book very well written and informative covering a broad range of relevant and important information concerning microbial food spoilage. Nevertheless, its high cost means that it is unlikely to be purchased by any individual, rather it would be a very good addition to an institutional or food industry library.

Overall it is an excellent publication and I recommend it highly.

Industrial Pharmaceutical Microbiology: Standards & Controls Editors: Norman Hodges and Geoff Hanlon. Euromed Communications Ltd, 2003. ISBN 1740-4975.

Price £325.00 Reviewed by Irina Barbolina The subtitle of this book is a ‘Practical international resource for all those working in the Pharma industry.’ Indeed, this book is a unique reference source for specialists dealing with the control of microorganisms in the manufacture of pharmaceutical, medical and medicinal products. The book does not teach microbiology but provides valuable help in understanding the industry specific requirements and regulatory expectations. Written as a manual, the book focuses on the microbiological issues of quality assurance, process control, product release, validation, safety and sterility.

The manual consists of 21 chapters dedicated to specific industry related microbiological topics. Each chapter is written by experts in this area, with most contributors having a strong industrial background from worldleading manufacture and consulting pharmaceutical companies.

The first two chapters look at the safety and quality assurance, and the role of qualified personnel in these procedures. Readers will find a guide to Good Manufacturing Practice and safety requirements in microbiology together with a clear summary of responsibilities. Each step is discussed with a respect to the existing procedures, requirements and standards.

Several chapters deal with the practical aspects of microbiological environmental monitoring, including: microbiology of water, chapter 4; microbiological environmental monitoring, chapter 6; prevention and elimination of microbial biofilm, chapter 8. This book provides clear step-by-step instructions for organising the correct procedures. It is impossible to give general advice for all practical situations, but this manual presents a

simplified guide in estimating the required scope of an environmental programme, taking into account the type of manufacture, suitable methods, validation techniques, and regulatory requirements.

To make this advice even more practical, the editors also include a detailed review of more complicated issues, such as cleanroom (chapter 9) and isolator technology (chapter 10). It provides an up-to-date approach in the decision making process by working back from the product or application towards cleanroom design, build and control procedures. One of the main advantages of the publication is that it considers not only existing standards, but also those which are likely to be adopted in the future, making the planning and decision making process easier.

Several chapters present an overview of existing and emerging methods and technologies with potential use in the pharmaceutical industry such as: enumeration and identification methods, chapter 5; methods of environmental monitoring, chapter 6; selection and use of cleaning and disinfection agents in pharmaceutical manufacturing, chapter 7; sterilization technologies, chapter 14; disinfectants testing protocols, chapter 16; measurements of biocides effectiveness, chapter 17. The overviews are not intended to give a detailed scientific description of listed methods nor products; they are rather concentrated on practical and regulatory considerations for selection and implementation.

Part of the book is dedicated to microbiological aspects of product quality and process control. Chapter 18 summarises general quality and regulatory requirements for biotherapeutics and manufactured products. Requirements, methodology, and validation of endotoxin tests and depyrogenation process are discussed separately in chapter 12. Anyone involved in quality assurance and process control will benefit from access to the following sections: Validation of aseptic processing and media fills, chapter 11; Biological indicators, chapter 13; Parametric release, chapter 15. The authors present each process in connection with practical principles, regulatory requirements, risk analysis, control, and validation.

The book concludes with a compilation of standards dealing with microbiological aspects of the manufacture and quality assurance. The complete list of relevant documents is supplemented by references to useful websites.

The book is clearly structured and very well written. The reference style allows easy navigation and efficient content searching. Each chapter is preceded by an introduction and depending on the topic, includes information about related regulatory guidance, design principles, and available methods or products. The clear and concise summaries and suggested reading list included at the end of each section are especially useful. This manual does not contain colour pictures but when necessary, clear, logical and helpful schematics, diagrams and illustrations are used to explain complex and difficult issues.

The most valuable feature of this book is a comparison of regulatory approaches and legislation requirements in Europe, USA and the Far East, and an explanation of philosophy behind these differences.

Such information will be extremely valuable in the industrial sector and gives this manual widely accepted international appeal.

The book is published in a loose-leaf format to allow it to be updated annually. Some updates and specialised supplements are already available to purchase.

The ‘Industrial Pharmaceutical Microbiology’ is an exceptional and unique publication in this field. This book is an extremely important and very practical collection of reference information for microbiologists, and other specialists in the pharmaceutical industry, medical devices and other related businesses. The editors combined in one volume the summary of all regulatory information necessary to start, maintain or improve microbiological control and assurance procedures.

Despite its institutional price, this book is value for money. I’m sure this manual will be actively used as a reference, navigation, and troubleshooting tool to help with many microbiological and nonmicrobiological aspects of pharmaceutical manufacture practice.

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news

The tale of a microbiologist and the media Richard James discusses the benefits of talking to the media

our policy on the media We will: ■ always do our best to provide facts, information and explanation.

■ if speculation is required, explain the rationale behind that speculation.

■ desist from hyping a story—whether it is the journalist or the scientist doing the hyping.

The Launch Symposium to open the Centre for Healthcare Associated Infections held at the University of Nottingham on January 5 2007 had a wider significance for me. Since 1975 I lectured on resistance to antibiotics and the problem of hospital-acquired infections (now called healthcare-associated infections, HCAI) and carried out research on killing of E.coli cells by protein antibiotics at the University of East Anglia. In the early years of my lectures antibiotic resistance was largely a scientific curiosity since a number of antibiotics were still available to treat every bacterial infection. As time passed bacteria became resistant to microbiology in the news multiple families of antibiotics and the rate of If you have any views on science in the media antibiotic discovery went into a long term decline. which you think should feature in this Gradually my lectures column, please send them to the Editor at: changed from ending lucy@sfam.org.uk. with what could be done to try to counteract the problem to why was something not being done.

Even the publication in 1998 of the comprehensive 7th report of the House of Lords Science and Technology Committee on “Resistance to antibiotics and other antimicrobial agents” did not relieve my sense of gloom and foreboding as the periodic press interest on HCAI, usually at election times, concentrated on the one small aspect of the problem that appeared to interest politicians,

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‘cleaner hospitals.’

In 2000 I moved to the University of Nottingham as Professor of Microbiology and was then giving lectures on antibiotic resistance and HCAI to BSc, MSc and medical students.

The term hospital superbug had entered into common usage as the incidence of methicillinresistant Staphylococcus aureus (MRSA) and later Clostridium difficile infections (and deaths) in hospitals rapidly increased. It soon became clear to me that there were many researchers in Nottingham, both in the University and the NHS, who were interested in many aspects of HCAI and that we could collectively make an impact to help reduce HCAI in UK hospitals. The idea of the Centre for Healthcare Associated Infections (CHAI) was born and I was happy to become its founding Director. The Centre has a unique breadth, with research staff from nine Schools and two research Institutes of the University as well as the University Hospital NHS Trust, that allows a unique holistic approach to research in healthcare associated infections.

The Launch Symposium of the Centre was the culmination of a lot of hard work and attracted a high quality audience of >200 delegates that included researchers from industry, universities and the NHS; NHS clinicians and Infection Control nurses; senior representatives of research funding bodies, the Department of Health and the Health Protection Agency; patient support groups; venture capitalists, solicitors and a member of the House of Lords. Before setting up the new Centre I remember informing

the Dean of the Faculty of Medicine that the Centre would attract considerable media interest because of the ‘superbug’ angle, but very close to the Symposium date we finalised the arrangements for Actress and TV personality Leslie Ash to be the Patron of the new Centre.

Media interest in the new Centre was exceptionally high and coverage of our research featured on national and local TV, national and regional radio stations and in newspaper articles.

The Director of Media Relations at Nottingham University was particularly pleased to see the Centre being the lead story on the BBC News at lunchtime that day.

With hindsight, I was grateful that the University had arranged in advance of the Launch Symposium for me to attend a course on giving media interviews that included the opportunity to practice recorded radio and TV interviews as in the last six months I have now given more than 70 media interviews. These ranged from in depth interviews with newspaper science correspondents, short recorded interviews with local radio stations, recorded items for the Trevor MacDonald Tonight program, live interviews on national TV, to a live TV debate with local politicians. You quickly learn that the contents of a 30 minute interview might be condensed to a 15 second soundbite in a TV news bulletin. I was particularly pleased with the contribution that researchers in the Centre made to the BBC Radio 4 programme ‘Warding off the germs’ that was presented with consummate professionalism and impressive background knowledge by Winifred Robinson. Most scientists would say that they get nervous just before giving a Conference lecture using content that they have prepared in advance, however the last few seconds waiting to be interviewed ‘down the line’ live on national TV when you do not know what the first question will be certainly gets the adrenaline going.

Summary Many scientists might question the value of being involved in media work and the time that it takes, but I believe that University researchers who obtain funding from public bodies have an obligation to discuss their research with the public. Research funding bodies are also increasingly looking for research grant holders to talk about their science in order to justify the continued public support for research. The media appearances gave me on behalf of CHAI a fantastic opportunity to move the debate on HCAI to other areas, such as the pressure of competing targets for reducing waiting lists on HCAI, and the need for universal screening for MRSA on admission to hospital, that extended beyond the usual discussions on the need for cleaner hospitals. I can at least say that we have

attempted to raise the complex issues with a larger audience in the hope that the UK develops an integrated strategy of measures that can help reduce the incidence of HCAI. Negotiations are underway with production companies that may result in some further opportunities to develop this theme in future TV programmes. I hope that other Microbiologists will look over the parapet and be willing to help inform the public about what they do and why it is important. The completely unexpected products of the media interviews were approaches from members of the public to make donations to the Centre, and an analysis in The Scientist of my use of ‘war metaphors’ when talking about waging war against HCAI.

Media tips 1. Register your research interests with the Science Media Centre who do a fantastic job in arranging press briefings for journalists and consulting scientists to help shape newspaper articles about science.

2. Get advice from your University, NHS or Society Media Relations office before a media appearance to check that you have permission to appear and to prepare you for the format, the need for a soundbite etc. If possible attend a course to help practice media skills; these are now often made available to grant holders by Research Councils. The Science Media Centre also run Introduction to the News Media sessions for scientists.

3. A live interview cannot be edited so it is your opportunity to set the agenda but it can be scary.

4. TV still likes the stereotypic image of a scientist in a white coat being interviewed in a lab so you need to carry out a documented risk assessment to facilitate this.

5. Media appearances usually lead to more invitations to appear so you may have to decide how much time you are willing to give up. For news and current affairs programmes there is no payment for giving an interview.

further reading Information about the Centre for Healthcare Associated Infections can be found at http://hcai.nottingham.ac.uk.

The extensive and very helpful Top tips for media work guide is available from the Science Media Centre — www.sciencemediacentre.org.

Richard James Director, Centre for Healthcare Associated Infections, Head of the School of Molecular Medical Sciences, University of Nottingham

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news

Med-Vet-Net’s 3rd Annual Scientific Meeting

med-vet-net Med-Vet-Net is a European Network of Excellence that aims to improve research on the prevention and control of zoonoses by integrating veterinary, medical and food science research.

Comprising 16 European partners and over 300 scientists, Med-Vet-Net will enable these scientists to share and enhance their knowledge and skills, and develop collaborative research projects. Med-Vet-Net officially commenced on 1 September 2004, and is funded to the value of €14.4 million for five years.

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Med-Vet-Net’s 3rd Annual Scientific Meeting, attended by nearly 200 delegates from 17 countries worldwide, was held from 27 to 30 June, near Lucca, Italy. The conference provided an opportunity for specialists in food borne disease from both clinical and veterinary backgrounds to exchange and share ideas as well as research methodologies that will enable greater cohesiveness and collaboration in the prevention and control of these diseases worldwide.

Sixty-seven talks and over 125 posters were presented at the conference on aspects of zoonoses related to epidemiology, including molecular epidemiology, risk research, detection and control, host–microbe interactions, and new and emerging zoonoses. A number of SfAM members were funded by Med-Vet-Net to attend the meeting, and SfAM Committee Member, Dr Mark Fielder, spoke on ‘The isolation of Staphylococcus aureus strains from raw chicken and pork from retail outlets in the UK.’ The meeting was opened by Valérie Baduel, Assistant Director of Agence Française de Sécurité Sanitaire des Aliments (AFSSA — French Food Safety Agency) and President of the Governing Board of Med-Vet-Net. She emphasized the importance of the multidisciplinary approach of the network to addressing problems of public health, as well as developing increasing connections with the European Food Safety Authority (EFSA). In addition, the President pledged to make sustainability of the network the priority of her mandate in the Governing Board.

Many of the keynote lectures looked at the new challenges to be faced in the field of zoonoses research, such as global climate change, avian influenza and vaccine development. The UK’s Chief Scientific Adviser, Professor Sir Howard Dalton, warned of a changing climate’s potential hazard to animal health and welfare, with new diseases having the potential to wipe out entire stocks of animals, in his lecture on the ‘Effects of climate change on infectious diseases.’ Dr Ilaria Capua, Head of Virology at Istituto Zooprofilattico Sperimentale delle Venezie, in her keynote ‘Four years of H5N1 — Where are we and where do we need to go?’ also spoke of a future challenge faced by the medical, veterinary and agricultural scientific communities; that of controlling avian influenza.

September 2007

The meeting’s focus on the future of both zoonoses research and Med-Vet-Net was continued in a panel discussion session with experts from academia, a public health institute, an international advisory body and, as consultant to the animal health industry, SfAM past President, Professor Peter Silley. The panel were tasked with answering the question: ‘What are the future challenges for zoonoses research and surveillance in Europe over the next 10 years?’ Ideas from these discussions will form the basis of developing strategies for the the long-term sustainability of Med-Vet-Net activities beyond the end of the European Comission funding in September 2009.

The first two Annual Scientific Meetings of the Network focused on networking and building

collaboration but, says Professor Diane Newell, Project Director, “By this third Annual Scientific Meeting of Med-Vet-Net our activities have matured and the outputs of various research networks are now coming to fruition. This was clearly demonstrated by the many presentations submitted from among our 22 Workpackages (research projects).” The quality of these research outputs is clearly recognized within the scientific community, with nearly a third of meeting participants coming from outside of Med-Vet-Net. In addition, it is of note that a number of delegates were from industry, highlighting the applied nature of the science carried out within the network.

Within Med-Vet-Net, a number of groups have come together to share information and develop

ideas in Special Interest groups. On the final day of the meeting, two of the groups had occasion to meet. The Emerging and Neglected Zoonoses Group held a workshop for all those, including non-Med-Vet-Net members, interested in the burgeoning area of MRSA as a potential zoonotic agent. The aim of the workshop was to develop, promote and take forward potential paths of research that can be moulded into a consortium bid for the upcoming deadlines for the European Union’s Framework Programme 7 (FP7) funding call. Chair of the meeting, Mark Fielder, said “Specifically, we need to identify potential calls that we can answer, decide if we need to lobby for a more specific funding call and identify the important or relevant branches of research that will aid a successful bid.”

In contrast, the Host–Pathogen Special Interest Group Session had a particular emphasis on the development of databases and networks. There was, however, also opportunity to discuss future collaborative funding opportunities in the area of host–pathogen interaction research.

Having sated their scientific appetites, meeting participants also had the opportunity to relax and have informal discussions while enjoying the panoramic views of the surrounding mountains on the conference-centre terrace or by the pool. A Tuscan-themed evening was held with a display of Gli Sbandieratori — traditional flag wavers — in medieval costume, followed by a dinner of regional dishes and local wines. On the final evening the Communications Unit hosted a lively quiz night, compered by Peter Silley, which was enjoyed by all, although there were a few disagreements over the answers!

The 4th Annual Scientific Meeting of Med-VetNet will be held in the seaside town of St Malo, in Brittany, France, from 11 to 14 June 2008.

We hope to have around 250 delegates, with many coming from outside the Med-Vet-Net network. SfAM members are most welcome to attend and submit abstracts. We are also now seeking sponsors for this meeting. À l’année prochaine!

abstracts A CD-ROM of abstracts from the meeting as well as many of the presentations is now available on request from the Med-Vet-Net Communications Unit: communications@medvetnet.org.

Jennie Drew Senior Communications Officer Med-Vet-Net

September 2007

information For more information about Met-Vet-Net, visit: www.medvetnet.org/ or contact Teresa Belcher on: +44 (0)1234 271020 For updates on the 2008 meeting in St Malo, France, please visit the 4th Med-VetNet Annual Scientific Meeting website: www.medvetnet.org/ mvnconf08

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ompetition is an essential part of scientific research culture. We aim to do “internationally competitive” research and funding is on a “competitive” basis. Many Laboratories seek to be first amongst equals because there are few accolades in being the second to make a discovery. Frequently this is difficult to achieve and highly competitive Laboratories frequently collaborate in order to be first together. This collaboration can be on a massive global scale. In general we thrive on competition and collaboration: the time to be worried is when nobody is interested in collaborating with you — it suggests that you are uncompetitive. This competition is a major driver for the success of science but there are occasions when it is unhelpful. I don’t intend here to review the major “spats” in the biosciences but will remark only that the current intense competition between two F1 racing drivers is not improving the morale of their team. Competition can be very positive but this upside can be replaced rather quickly by a negative effect if the Richard Dyer discusses the pros and cons competition is inappropriate.

There are many of competition within the biosciences organisations active in the biosciences. There are at least 80 Learned Societies and about half of these have come together in the Biosciences Federation (BSF). But these Societies are together whether or not they are part of an umbrella organisation: They are “together” in Team The Biosciences Federation is a single authority Bioscience. “Together” the representing the UK’s biological expertise, Team faces competition for providing independent opinion to inform public money, students, specialists policy and promoting the advancement of the and infrastructure from the biosciences. arts, the humanities and other branches of science and For further information visit: engineering. Team Bioscience http://www.bsf.ac.uk/default.htm has created a competitive internal market and has sometimes lost sight of the real competition.

For example, quite a few Learned Societies have staff working to get teaching resources into schools. My impression is that they are informed and dedicated people doing excellent jobs. But over and over again I hear that part of the motivation is to expose young people to the words that describe their Society - “microbiology”, “endocrinology”, “physiology”, “biochemistry”, “ecology” and probably all the other ologies! This is crazy because it is impossible for a single school to take on all these different resources, focussed on a separate sub-discipline of the biosciences. The internal market means that “microbiology” is successful if it gets more “hits” than “endocrinology”. Do we think this form of internal competition is helpful? What we really need is more students as a whole thinking of the biosciences as a career and a bigger and better qualified pool of young bioscientists. When we achieve these goals both the microbiologists and the endocrinologists will get more recruits – as will

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everyone else.

What do our competitors do? The Royal Society of Chemistry is also active with young people and looks to create teaching resources. However these are not focussed on “analytical chemistry”, “synthetic organic chemistry”, or any other subset of the subject of chemistry. Their efforts are exclusively focussed on chemistry because first and foremost they want to encourage more young people to become chemists. Their policy is wise.

Where else is there an internal market? Certainly in policy work! Currently it is quite common for the BSF to respond to an enquiry and find that several Member Organisations have produced their own response. I shall work harder to ensure that competition with Member Organisations is reduced but I can’t influence a decision of members to “do their own thing”. I am convinced that this is ineffective and therefore a waste of money.

Somehow the BSF must increase its catalytic capacity to get a unified voice for the biosciences on the major topics that impact on the future health of our discipline.

More broadly, we must identify those areas where Team Bioscience has created an internal market that acts as a detriment to success in the external market. Solutions are possible for both the examples given above. The BSF was established to help provide these solutions and I detect the landscape is changing in a way that will allow this to happen more readily than in the past. There is increasing awareness amongst Member Organisations, and their membership, that a structure for the Biosciences that seemed to work in the mid twentieth century is no longer appropriate today. Several weeks ago I was at an open meeting where Sir David King asked in his talk if the fractured Bioscience landscape was “fit for purpose”! The answer depends on which purpose you consider. If the purpose is for microbiologists, ecologists, physiologists or plant pathologists to meet to talk about their work, the answer is yes. In the practical science context, Societies are successful “special interest groups.”

The “yes” is qualified because young people don’t always relate to the disciplines around which some Societies are built and this may be a future problem for them. However, it is very much harder to answer “yes” if the “fit for purpose” question refers to outreach and to engagement with local, national and European politicians and opinion formers.

These areas need Team Bioscience.

Some competition between Learned Societies will remain for the foreseeable future. However, I do believe there is an increased wish for us all to work together whenever appropriate and possible.

This wish was rather theoretical two years ago: today I see much more desire for its implementation. I am confident that Team Bioscience will be built. Its shape and final structure is a little uncertain but I am confident of the outcome because the need is so great.

Richard Dyer Chief Executive Biosciences Federation

meetings

information For more information about the Society’s meetings please visit the website at: www.sfam.org.uk You can also find details of next years’ Winter Meeting on page 29 of this issue of Microbiologist

Summer Conference 2007 Report Microbiology of Fresh Produce Park Plaza Hotel, Cardiff, Monday 2 to Thursday 5 July 2007

Introduction

Session 1

he summer conference 2007, Microbiology of Fresh Produce kicked off with a fascinating talk from Dr Peter Green to honour the work of scientist Lewis B Perry held at the National Museum of Wales in Cardiff. The presentation entitled: “Bacterial anti-cancer vaccines: a science frozen in time” covered the work of scientist William Coley who was a young doctor in the late 1890s and early 1900s. His work investigated the observation that many cancer patients, who had encountered acute infection at some point during their illness, were subject to significant regression or complete recovery from the cancer after the infectious episode.

The development of a bacterial vaccine known as Coley’s toxin followed — a radical new treatment for cancer. Dr Green explained details of this remarkable work and how it fell out of favour with the introduction of radiotherapy and later chemotherapy. This thought-provoking talk was among the many topics of conversation at the reception which followed, giving delegates a chance to meet each other and chat over drinks and canapés. Later that evening, delegates met at the venue for the remainder of the conference, the Park Plaza hotel, for an entertaining pub quiz hosted by Honorary General Secretary, Dr Anthony Hilton.

The main conference opened with an overview by Mike Doyle (University of Georgia) on “The problems with fresh produce.” He concentrated on the situation in the USA where microbial contamination of fresh fruit and vegetables has been identified as a leading cause of food borne illness. He discussed the recent USA outbreaks of E. coli O157 associated with spinach and lettuce and Salmonella from lettuce, tomatoes and melons. These outbreaks occurred mainly in the autumn and he predicts that there will be more.

He discussed the trace-back work to identify the source of the infections and described the research being carried out on routes of infection, survival of pathogens on and around crop plants, prevention and cleansing. He finished by suggesting that risk assessment studies are useful tools in developing risk management strategies for the produce industry.

The rest of the morning was taken up with talks on the “Organisms and the Plant.” The first was given by Bill Keevil (University of Southampton) on “Zoonotic pathogen interaction with the microflora of the growing crop phylloplane.” Bill showed that crop plant surfaces are naturally colonised by a wide variety of microorganisms which derive from soil, wind blown dust, manure, rain splash and faecal contamination from animals and birds. He concentrated on how to measure and follow the attachment of bacteria to leaf surfaces and described elegant real time, in situ electron microscopy that circumvented some of the artefacts caused by drying and coating.

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Lucy Harper

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He showed how bacteria adhere and integrate into existing biofilms on the surface of leaves and demonstrated that curli enable Salmonella to attach better than E. coli.

Bill was followed by Joanna Heaton (Lancaster University) who discussed “Factors affecting the bacterial contamination of salad vegetables and the persistence of enteropathogens in the phyllosphere.” Joanna is in the final year of her PhD and this was her first lecture at a major conference. She presented data from a year-long study of the microbial contamination of lettuce and other salad crops at point of sale in various outlets in Lancaster, concluding that on the whole, fresh produce is within the EU guidelines for E. coli and Listeria — in line with the UK wide Food Standard Agency surveys. Measurements of field crops showed that growing lettuce carries large, but variable, populations of epiphytic bacteria. However, lettuce grown in polytunnels under different UV regimes (using spectrally modifying plastics) showed a reduction in the numbers of bacteria on lettuce exposed to UV-B. She then concentrated on the dose response to UV of free-living and attached Salmonella, and discussed the methods used to grow sterile plants for attachment studies. She concluded by suggesting that the normal epiphytic population may be required for enteropathogens to attach to leaves.

After a coffee break, Christine Dodd (University of Nottingham) discussed the “Non-culture based approaches to examining the microflora of salad vegetables.” She explained that ready-to-eat salad vegetables support the growth and or survival of food borne pathogens and that the ways in which crops are grown, washed and stored affect the levels of contamination. Washing and disinfection rarely produces more than a one or two logs reduction in microbial loading and leafy vegetables and herbs have structures which permit attachment. Attached microorganisms are more resistant to disinfection than non-attached. She showed that a range of molecular-based methods are useful in following washing and disinfection decontamination processes. She also emphasised the role that the normal epiphytic microflora might play in bio control and the reduction of attachment by pathogens.

Christine was followed by Tim Brocklehurst (Institute of Food Research, Norwich) on the topic “Measurement and modelling of the attachment of bacteria to plant surfaces.” Tim explained that although bacteria are found on both the upper and lower surfaces of crop plant leaves, their attachment and subsequent growth is much greater on cut surfaces. He described the three stages of bacterial attachment to cut surfaces: a rapid attachment phase; consolidation, which might involve extracellular polymer production; and growth. Tim went on to describe their stylish molecular-based work to elucidate the mechanisms of early attachment and presented models that can be used to predict the extent of colonisation of cut fruit and vegetables in a range of conditions.

The last talk of the morning session was given by Amy Charkowski (University of Wisconsin) on “Erwinia soft rots — ecology and genomics of broad host range plant pathogens.” This represented a change from looking at human pathogens on crop plants to one looking at pathogens of the crop plants themselves. Soft rot erwinia, now called Pectobacterium and Dickeya, are enterobacterial plant pathogens that cause soft rot, wilt, stem rot and blackleg in a 24

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range of crops, including carrots and potatoes. They are found world wide on plants, in water and in insects. Once decay of a crop begins, human pathogens such as Salmonella can multiply in the tissues. It is not known whether this is a real health risk. After rot sets in there are no effective control measures, so presently growers have only sanitation to rely on. Research is currently under way to develop bio control methods, competitive exclusion and resistant crops. Amy also described the genome projects and advances in topics such as phytotoxin synthesis and virulence in Pectobacterium and Dickeya, the detailed molecular methodology of which is in stark contrast to the other part of her job, which is to administer the Wisconsin Seed Potato Certification Program and run the University potato farm.

Keith Jones, Lancaster University

Session 2 Mycology is a frequently neglected area of food microbiology and one that is particularly important in the determining the quality of fruit and vegetables. Maurice Moss gave a splendid overview of the relationship between fungi and these commodities, focussing particularly on how this can result in spoilage but also addressing some of the safety and regulatory issues with respect to patulin in apples and apple juice and tenuazonic acid produced by Alternaria alternata in tomatoes.

Quarantine pathogens are those which can affect domestic crop production and therefore need to be excluded from the UK. To do this effectively we require efficient and accurate methods to detect the organisms in imported produce. Simon Weller from the Central Science Laboratories, York, described the work he and his colleagues have done in perfecting molecular detection techniques based around Polymerase Chain Reaction (PCR). Such techniques count for nothing if it is not possible to extract DNA reliably from plant tissues and his talk also described some rapid and simple DNA extraction

techniques that can be used in the field.

The meeting then moved on to consider “Public Health Aspects” relating to fresh produce and Christine Little from the Health Protection Agency reviewed the data relating to microbiological quality of prepared salads in the UK. Compared to the United States, outbreaks associated with these products in the UK form a relatively minor part of total food borne disease and surveys of retail products have shown that the incidence of food borne pathogens in them is relatively rare. However their consumption is increasing and there have been a number of high profile product recalls in recent months. The power of modern traceablility and typing were illustrated by one example where a particular serotype of Salmonella isolated from lettuce was traced back to the field where it was grown and carriage by field lizards living there.

The use of farm wastes such as manure and slurry are an obvious potential route for the transmission of zoonotic pathogens to ready to eat produce and Michael Hutchison of the University of Bristol described the results of a large programme he participated in to determine the rate of survival of pathogens when disposed of to land. The way in which the waste is applied can have a major impact on survival and this can give rise to some difficult decisions. It seems that methods which minimise the production of greenhouse gases such as methane also give better survival of pathogens. How these conflicting objectives should be prioritised or resolved is a thorny question for risk managers.

Martin Adams, Meetings Secretary Tuesday ended with a lively Trade Show with a Caricaturist and an anagram quiz which maintained the theme of the conference: Fresh Produce. Each of the Trade stands provided a prize for this competition with winners receiving gifts including a well known brand music player and a bottle of champagne!

Session 3 The science underpinning the microbiology of fresh produce should inform us not only about the problems and principles involved in the colonisation of foods, but also about potential intervention strategies. Session three built on the previous sessions to do just that.

Jay Hinton from the Institute of Food Research in Norwich related advanced molecular microbiology approaches. His expertise enables interrogation of the entire genome of sequenced organisms following an environmental change. So, for example, he could use his approaches to identify the genes of Salmonella or E. coli that are upregulated when the bacteria colonise the surfaces of fruit or vegetable tissues.

This essentially identifies the bacterial competence required for colonisation. Equally significant would be the changes to an organism’s gene expression during its transit through the gastrointestinal tract that results in illness (which is Jay’s main area of research). An interesting issue raised by the audience is the behaviour of the bacterial genome when the organism moves from its animal host onto plant surfaces and results in contamination of the produce in the first instance.

Movement of bacteria between hosts and between humans and plants was the topic pursued by Eshwar Mahenthiralingam from Cardiff University. He focussed on opportunistic pathogens, including Pseudomonas aeruginosa, but principally the Burkholderia complex, that cause serious disease in compromised individuals. Eshwar is also exploiting molecular microbiology using Signature Tagged Mutagenesis to construct mutants that differ in their virulence, but also in their ability to adhere to the rhizosphere. Each of these first two of our presentations raised the prospect of molecular biology providing clues to the mechanisms of attachment of bacteria to fresh produce that could be exploited in order to intelligently design intervention strategies.

Our next two speakers identified the need for such strategies. John Bassett from Unilever UK described how the potential for illness arising from consumption of contaminated fruits and vegetables could be systematized by application of Risk Assessment methodologies. John described his approach to constructing a qualitative risk assessment. This is an overarching approach to the microbiological safety of fresh fruits and vegetables, and served to identify some of the gaps in our knowledge. The Session continued with “Intervention Strategies for Control.” Robert Gravani from Cornell University in the USA described the huge role that Good Agricultural Practices have to play in minimising microbiological problems with fresh produce. Robert has a program of education and training directed towards the US produce industry. He enthralled the audience with anecdotes that were at once amusing and serious. It is clear that growers have an enormous contribution to make in minimising contamination in the field and Robert’s description of the American approaches included not just the conventional information sheets, but also short movies and comic book stories to get the messages across.

Finally, we were introduced to potential physical methods for the decontamination of produce. Stephen James of the Food Refrigeration and Process Engineering Research Centre in Langford described the inadequacies of some existing September 2007

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chemical and physical methods for decontamination before outlining potential alternatives. It is quite clear that engineering approaches have a place if designing improved processing. What we should aim for is an amalgamation of all of the approaches described in this Session, so that we identify the contamination bottlenecks, the mechanisms used by the bacteria to successfully colonise plant tissues, and then design suitable strategies for intervention.

Tim Brocklehurst, Institute of Food Research

Afternoon Session In the afternoon session that ran up to the W H Pierce Prize and annual general Meeting we were offered eight papers including five student papers which this year were chaired by an active student member, Andrew Hall. Jorge Gutierrez and co-authors from the Dublin Institute of Technology presented their evaluation of the antimicrobial activity of plant essential oils against food borne pathogens and spoilage bacteria. They presented various data that suggested that Essential Oils (EOs) derived from oregano and thyme had the highest antimicrobial activity against Listeria monocytogenes and Pseudomonas spp. while other EOs studied, including basil, caraway, rosemary and sage, and parsley had moderate or no activity. These data would seem to have great applicability in the food processing environment although some of the first questions to be asked were about the flavouring effect on food products and how the authors envisage the application of EOs in the food processing environment.

Sue Jones from Wessex Environmental Microbiology services presented an interesting and insightful comparison of data from two years’ microbiological survey of processed fruit and vegetables from UK suppliers and producers with data from imported sampled received by air or at container ports.

In 95% of cases, the microbiological quality of fruit and 26

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vegetables was satisfactory. However, not only did this presentation give insight in to some of the aspects of the microbiology of world food markets, with data on banana leaves and a few other vegetables with which most of the audience were not familiar, but the conclusions of the authors, that the low prevalence of pathogens found does not concur with the epidemiological data reporting fruit/vegetables as vehicles of known food poisoning outbreaks, served as a reminder that industry and enforcement agencies still need rapid real time microbiological analyses to be able to provide public health protection in this rapidly changing part of the economy.

Our last offered presentation before the student session, from Adrian Peters of University of Wales Institute, Cardiff, took us to the microbial world of the drink vending machine, which, perhaps unsurprisingly, presents the industry with considerable problems. While correct operational management of machines normally ought to minimise microbial loading of drinks, the authors presented data that characterised the build up biofilms which included species of Pseudomonas, Aeromonas and Stenotrophomonas, as well as occasional coliforms. The latter highlighted the importance of proper hygiene management and it is clear that the work presented is contributing to a greater understanding of the maintenance and safe functioning of vending machines.

Karen Stanley, Sheffield Hallam University

Students Presentations This year’s student session saw five excellent presentations on a diverse range of microbiological topics. Each student gave a 10 minute presentation and fielded numerous questions from the audience. The session began with James Cass’ talk on enteric pathogens in biosolids amended agricultural soils. This was followed by a report by Katie Fisher on the use of citrus essential oils against

Enterococcus spp. Christopher Ibenegbu described the optimisation of xylanase production by a mixed culture of Trichoderma sp. and Aspergillus niger during solid substrate fermentation of brewery spent grains. The prize for best student oral presentation was awarded to Victoria McCune for her presentation on the Development of a multiplexed PCR-microsphere array for the rapid detection of gastrointestinal pathogens from foods. The session was concluded by Haruna Musa who spoke on microbial volatile organic compounds (MVOCs) as indicators of mould presence in homes and the evaluation of the possible cytotoxic and genotoxic potentials of selected MVOCs. Andrew Hall, University of Wales Institute, Cardiff

This session concluded with an interesting talk from the winner of this years’ Pierce Prize, Dr Dennis Linton of the University of Manchester (see page 13). His presentation, entitled: “Phase variation and the ever-changing surface of Campylobacter jejuni� covered his work into the functional characterisation of phase variable genes of C. jejuni. These are located at loci involved in the biogenesis of components of the bacterial cell surface, including the flagella. The understanding of the structural consequences of this phase variation, in particular that associated with the formation of flagella, could give us greater understanding in the control of C. Jejuni.

The day ended with a chance for delegates to mingle and talk about events of the day at the conference dinner. This was preceded by, for those who wished to attend, a tour of the Millennium Stadium, Cardiff. After a delicious spread, the winner of the student oral presentation, Victoria McClune, was presented with her prize by David Wareing of Invitrogen and the student poster winners, Kiera Considine (1st prize), George Aboagye (2nd prize), and Shadlia Matug (3rd prize) received their prizes as presented by Vicky Johnson of Wiley-Blackwell.

Session 5 The final session of the meeting continued with the theme of intervention strategies for the control of microbial contamination and then went on to explore some industrial perspectives. Christian James of the Food Refrigeration and Process Engineering Centre at the University of Bristol discussed the alternatives to chlorine for removing microbial contaminants from harvested produce. Antimicrobial treatments must ideally remove contaminants without changing the intrinsic nature of the product in order to increase consumer safety and prolong shelf life. Washing alone is capable of achieving up to a 2-log reduction in contaminants and so the use of chemicals must clearly surpass this. However, most antimicrobial chemicals exert some harmful effects or may impair colour, flavour or taste of the food. The most commonly used chemical agent is chlorine because it is cheap and brings about reductions of greater than 2-logs. However, its safety profile is such that other compounds are now being sought. Dr James described work done on the washing of fresh herbs with chlorine whereby an understanding of its limitations enabled the company concerned to reduce both the concentration used and the time of exposure. Other chemical treatments described were ozone, lactoperoxidase, and organic acids, particularly lactic acid and vinegar. Vinegar was more efficient against coliforms than chlorine but was much more expensive and also had little effect on non-coliform contaminants. In conclusion, no agents tested were able to eliminate all surface microorganisms and there were still concerns over safety, product quality and environmental impact.

The increase in consumption of ready-to-use fresh fruit and vegetables has led to a nearly 70% increase in produce packed in modified atmosphere conditions since 2001. Gail Betts of Campden and Chorleywood Food Research Association (CCFRA) defined modified atmosphere packaging (MAP) as packing of produce in an atmosphere different from air. The gases used were generally oxygen, carbon dioxide September 2007

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and nitrogen and this has been shown to be a very effective preservation technique particularly for chilled perishable foods. The shelf-life of MAP produce can be up to twice as long as air packed samples due to inhibition of aerobic microorganisms but many spoilage organisms and pathogens can grow under modified atmospheres. In addition, natural produce will continue to respire after packaging and it is important to balance the modified atmosphere gas mixture with the packaging materials used and the natural respiration rate of the produce. Good hygienic practices during growing, harvesting washing and packaging should also be in place to minimise the potential for pathogen contamination.

Debra Smith, also from CCFRA reported on a three year study attempting to identify the relative importance of different vectors of cross contamination in the processing of ready-to-eat produce. The vectors studied included equipment surfaces, operative’s hands, wash tank water and the air. The study included a detailed investigation of the process of food

administrative requirements for food businesses and neither do they lead to additional costs. A number of myths surrounding the Regulations were dismissed. A key point is that food safety is neither guaranteed nor controlled by microbiological testing and it is imperative to be able to demonstrate GMP and functioning HACCP-based systems.

Free downloads on guidance related to MCR are available from www.chilledfood.org Demand for organic produce has increased in recent years and organic farming standards recommend the use of regular manure inputs and prohibit the use of chemosynthetic pesticides. Carlo Leiffert of the Nafferton Farming Group, University of Newcastle, evaluated the risks of enteric pathogens entering the supply chain as a result of this strategy. A number of advantages of regular organic matter inputs (manure) were highlighted particularly with respect to soil content and fertility. Data were also presented to show health benefits associated with rearing livestock outdoors for

preparation during factory visits together with laboratorybased experimental work on agar to agar microbial transfers.

An interesting observation was that the use of gloved hands resulted in a higher percentage transfer rate of microorganisms than bare hands. Overall, the vectors were ranked in order of importance with surfaces being the most significant followed by air; then bare hands; gloved hands and finally wash liquids. The general conclusions were that each process needs to be individually assessed and repeated microbial sampling of vectors during production is essential to determine a true idea of loading. The laboratory studies showed that vector loading was more important that percentage transfer and therefore the prime objective should be to minimise initial bioburden. Exposure assessment could be used as part of the Hazard Analysis and Critical Control Point (HACCP) pre-requisites and HACCP food safety management systems.

The EU Microbiological Criteria Regulations (MCR) came into force on 11 January 2006 and have given rise to concerns in the food industry. Kaarin Goodburn of the Chilled Foods Association attempted to clarify the Regulations and show that they do not bring any new obligations or new

extended periods. Despite the potential risks of transferring pathogens to vegetable, salad and fruit crops it was shown that a HACCP-based approach could minimise these risks.

David Kennedy of Bakkavor Ltd., explained that salad crops are grown around the world on a massive scale to supply the UK market. These are not grown under laboratory conditions but in fields and employing local labour. While there is a perception that this carries high risk the reality is that since 1992 about 2.5 billion bags of salad have been produced in the UK and only two possible cases of food poisoning have been linked. The practicalities involved in the production of salad crops around the world were explained to highlight the difficulties of managing microbial quality. Risk assessments identified key issues such as irrigation, manure, staff hygiene and post harvest processes which were evaluated during annual technical onsite audits. Despite the difficulties the implementation of appropriate risk management and processing systems make the supply of fresh fruit and vegetables in the UK retail chain one of the safest in the world.

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Geoff Hanlon, University of Brighton

Winter meeting 2008 A one day meeting on

Quality Assurance and Alcoholic Beverages Royal Society, Carlton House Terrace, London Wednesday 9 January 2008

CPD

For the latest programme please visit us online at www.sfam.org.uk

ACCREDITATION

APPLIED FOR

including

The Denver Russell Memorial Lecture Prevention of biomaterial-based medical devicerelated infection Delivered by Sean Gorman, School of Pharmacy, Queen’s University of Belfast

September 2007

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Programme

Quality Assurance and Alcoholic Beverages

10.00-10.30 Tea, coffee and registration

14.45-15.05 Tea and coffee

Chair:

15.05-15.35 Internal quality auditing in clinical and food microbiology laboratories Ian Sharp, Quality Systems Unit, HPA Centre for Infections, Colindale.

Margaret Patterson

10.30-11.15 The Denver Russell Memorial Lecture: Prevention of biomaterial-based medical device-related infection Sean Gorman, School of Pharmacy, Queen’s University of Belfast.

11.15-11.45 Quality assurance in the university laboratory — is it necessary?

Sandy Primrose, Business & Technology Management, High Wycombe.

11.45-12.15 Systems approaches to optimise lager fermentations Katherine Smart, University of Nottingham.

Session B. The Microbiology of alcoholic beverages Chair:

Martin Adams

13.15-13.45 The microbiology of Belgian lambic beers Hubert Verachtert, University of Leuven, Belgium

12.15-13.15 Lunch

Session A. Quality assurance and accreditation issues in microbiology Chair:

15.35-16.05 Living with the known unknowns — Uncertainty of measurement in food and environmental microbiology Melody Greenwood, Wessex Environmental Microbiology Services, Southampton

13.45-14.15 The malo-lactic fermentation in the maturation of wine and cider Bob Lovitt, University of Swansea

Andrew Sails

13.15 -13.45 Getting your food and environmental laboratory accredited — some common problems and misconceptions Delia Cope, UKAS, Middlesex.

14.15-14.45 Bacterial and yeast contributions to Scotch Whisky production Derek Jamieson, Heriot Watt University 14.45-15.05 Tea and coffee

13.45-14.15 Quality control in molecular diagnostics Paul Wallace, General Manager, Quality Control in Molecular Diagnostics, Glasgow.

15.05-15.35 Wine yeast — transcriptional profiling of wine fermentations Bruno Blondin, INRA, Montpelier, France

14.15-14.45 Quality control in food, water and environmental microbiology Julie Russell, Head of the Food and Environmental Proficiency Testing Unit, HPA Centre for Infections, Colindale.

15.35-16.05 The role of the barley microflora in premature yeast flocculation during fermentation.

Barry Axcell, SAB Miller, South Africa

For the latest information please visit the website at: www.sfam.org.uk/winter_meetings.php

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September 2007

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WINTER

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9

JANUARY

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Left: the Director of IVI, John Clemens, visiting the Calcutta IVI field site. Inset below: the author and his collaborator Dr Sam Kariuki working in Sam’s laboratory in Kenya

Gordon Dougan presents a highly personalised view of the field of vaccinology and how it is developing at a global level

Getting vaccines to those

A

s all good microbiologists know infectious diseases are still a scourge of mankind. This is despite the fact that we know a great deal about how communicable diseases spread and have had therapeutic tools such as vaccines, antibiotics and diagnostics available to us for decades. As we learn more about the genetic make up of pathogens we realise that we are not dealing with a static pool of organisms but rather a rapidly evolving and dynamic challenge. Despite this challenge humans have been able to shape the communicable disease burden and have made enormous progress in some areas in terms of effective disease control.

One of the consequences of the application of disease control is that the global distribution of infectious diseases differs dramatically around the world. In most developed countries the epidemic spread of pathogens in the general population has been curtailed to a great degree. Here pathogens occupy particular ecological niches such as hospitals, where there is a critical concentration of 32

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susceptible individuals and specialised transmission routes.

However, in poorer regions we still have high levels of the classical epidemic diseases such as measles, pneumonia and diarrhoea as well as the emergence of new epidemic threats such as HIV/AIDS. The disparity between the rich and the poor is due to the methods and intensity of the application of both classical public health controls (improved hygiene and infrastructure) and therapeutic controls such as vaccination programmes. Vaccines are clearly one of the key infectious disease control tools we have. Consequently, here I would like to use the example of vaccines to illustrate how the disparity between the rich and the poor is sustained and to highlight how people and organisations are trying to close this gap.

Vaccination programmes, a brief history The scientific concept of vaccination was introduced by Jenner at the end of the 18th century but the first intense period of vaccine development occurred a century later starting with the work of Pasteur and continuing into the first

Below: the IVI building in Korea donated by the Korean government. Inset left: images of children in the Calcutta slums

Bibliography â– GAVI Alliance: http://www.gavialliance.org â– International vaccine Institute: http://www.ivi.org

that need them most part of the 20th Century. At this time, vaccines such as those targeting diphtheria, tetanus, tuberculosis (TB) and typhoid were invented. The processes of vaccine manufacture evolved during the mid-20th Century and both state owned and private pharmaceutical companies began providing products for use by the general public. In the 1950s in the developed world classical vaccines such as polio and whooping cough were licensed and these formed the cornerstones of public vaccination programmes that frequently operated at a national level. Vaccines manufacture became a regulated process adhering to World Health Organisation (WHO) and national guidelines for quality and reproducibility. There was an inevitable economic burden associated with core vaccination programmes but these were subsidised by governments and in some instances vaccination became a legal requirement. This was all well and good in developed countries but in many parts of the world national vaccination programmes were never implemented. Even when they were, vaccines were often made by local manufactures with a

variable application of quality control. Nevertheless, at this point there were many manufacturers making generic vaccines that could potentially supply local needs.

It is a sobering thought that when I was brought up in the 1950s and 60s in an environment of increasing vaccine coverage (personally benefiting enormously from the stability this brought), my equivalent birth cohort in other parts of the world was still exposed to the ravages of vaccine controllable illnesses. At this time point it is estimated that less than 5% of the children in the world were receiving even the basic children’s vaccines such as whooping cough, measles and diphtheria. Thus, diseases that were disappearing from developed countries continued to ravage developing countries. In response to this alarming statistic WHO launched an initiative, the Expanded Programme of Immunisation (EPI), to target the common vaccination preventable diseases, including diphtheria, tetanus, whooping cough, polio, measles and TB. This was arguably one of the most important health campaigns of the 20th Century and by September 2007

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Figure 1. A generalised overview of the way in which vaccine development has evolved over the years since Jenner

approach from a UK perspective as the health system was only prepared to meet the costs of its own requirements (indeed it may be ethically difficult to justify including vaccine coverage of a low incidence bacteria).

The preceding discussions highlight several points (i) even with EPI at least 20% of children in the world were not being vaccinated; (ii) new vaccines were not getting to even more children. They were either too expensive or did not readily fit into the EPI schedule; (iii) The vaccine business was largely in the hands of multi-nationals who, although dramatically moving the whole field of vaccine development and quality forward, were unable or unwilling to satisfy the global need.

Enter the philanthropist and global agencies

the late1980’s statistics indicated that almost 80% of the 130 million children born each year were vaccinated before their first birthday.

The Impact of new technologies on vaccine development After the 1960s we entered a fallow period where relatively few new vaccines were developed. This was, in part, because we had reached the limitations of the then available technology. Many pathogens, such as Hepatitis viruses, were difficult to grow in vitro at levels suitable for antigen production and for other diseases the protective antigens were not known. One of the consequences was that the whole vaccine area became unattractive to industry as patentprotected products disappeared from the pipeline and one by one the manufacturers stopped production. Ironically, this was at a time where the science of vaccinology was about to take off through developments such as genetic engineering and monoclonal antibodies. By the end of the 1990s many local producers had gone out of business and the whole industry was in the hands of a handful of multi-national companies and smaller companies. A consequence of this was that knowledge of how to make vaccines was lost from many parts of the world. Multi-nationals respond to the needs of shareholders and focused on the rewarding markets of the USA, Europe and Japan while generally ignoring the needs of developing countries.

During the past twenty years a range of new vaccines have been invented using new technologies. Such vaccines include Hepatitis B, Pneumococcus, Haemophilus, Rotavirus and more recently Papillomavirus. However, many of these vaccines were developed under patent and are consequently very expensive. Even potentially cheaper vaccines, such as the Vi polysaccharide vaccine for typhoid were sub-optimal for use in developing world children or were simply not taken up by EPI or any other route. Once additional spin off effect of the focus of manufacturers was that even when some novel vaccines were developed (e.g. for Meningitis) they were targeted to the needs of the rich customer. In the case of meningitis the vaccine targeted the Meningitis C antigen type circulating in the UK but did not include a Meningitis A component needed in Africa. This was a perfectly valid 34

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Due to a combination of factors at the turn of the century renewed interest began to emerge in global vaccination programmes. Stimulating factors included (i) the recognition of the scale of the HIV/TB/malaria global burden (ii) recognition of the impact of poverty on political and economic stability and (iii) the concept of the global village effect where nobody is immune to the threat of a potential emerging pathogen or a global pandemic. Infectious diseases were discussed at G8 meetings and promises and schemes began to appear. A critical player was the Global Alliance for Vaccines and Immunisation (GAVI Alliance). The GAVI Alliance is a public-private partnership focused on increasing children's access to vaccines in poor countries. Partners include the GAVI Fund, national governments, UNICEF, WHO, The World Bank, the Bill & Melinda Gates Foundation, the vaccine industry, public health institutions and nongovernmental organisations. GAVI and related organisations have stimulated interest in creating a ‘market’ whereby it is attractive for manufacture’s to supply vaccines to the poorest individuals. To this end a global fund is being built and the concept of ‘advanced market commitments’ has been developed. The aim is to purchase manufactured vaccine lots prior to their distribution to the end user, hence guaranteeing a market (push/pull effects) to stimulate growth in the sector. These ideas aim to stimulate vaccine activity and help governments in the poorest countries introduce effective national vaccination programmes.

Sustainability — a personal view As someone who worked in the vaccine industry in the period of decline in the 1980s it is heart-warming to participate in the new initiatives to get vaccines to those who need them. The work of the global agencies is providing essential finance and impetus to the area but how can this be sustained long term? Clearly the multi-nationals have the expertise and know how to make vaccines of the highest quality. However, they have to ensure that profitability is at the top of their agenda. For example, they cannot be expected to develop new vaccines against neglected diseases such as schistosomiasis or enteric infections mainly localised in poor regions. They also generally are not motivated to adapting vaccines for use in tropical regions (targeted antigen-types, creating heat-stable formulations etc). Such approaches might be more attractive to local or specialist enterprises provided they can operate at the highest quality level. However, we have already stated that many manufacturers disappeared from the developing world in the past three decades. Hence, there is a dearth of expertise in these areas. Encouragingly,

Figure 2. Dates that key vaccines were introduced with information on how they are used today

More recently the pharmaceutical company Novartis has announced the establishment of the Novartis Vacines Institute for Global Health in Siena, Italy whose aim will be to apply the state-of-the-art technologies to vaccine development against neglected disease targets and transfer technology to manufacturers who will supply countries where there is an endemic need. We can expect more initiatives of this type in the future.

The role of the individual

new vaccine manufacturers are beginning to re-emerge in different countries of the world, partly in response to supply limitations. Examples include India and Cuba, where many new manufacturers are now competing. Others are appearing in Asia and South America, although there is still a need in Africa. Although these new players are currently concentrating on generic vaccines if they survive they will theoretically be able to formulate vaccines targeted to disease problems in their immediate area. Remember, that many of these diseases have now become tractable due to new technologies.

Another encouraging development has been the emergence of specialist scientific organisations dedicated to improving the delivery of vaccines to the poorest using practical approaches. An example of such an organisation is the International Vaccine Institute (IVI) in Korea, an International Organization established at the initiative of the United Nations Development Programme and the WHO. The mission of the IVI is to contribute to the reduction of vaccine preventable diseases in developing countries by collaborative research that generates the evidence needed for rational introduction of new vaccines, supported by programs of basic and applied laboratory research, product development, training, and technical assistance. To this end IVI has been performing disease burden studies linked to demonstration projects where vaccine efficacy is measured in an endemic area where the most needy live. Disease burden studies provide proof of the need for vaccination (often lacking in the eyes of politicians who have to juggle limited budgets) and can provide information on real cost effectiveness. The institute’s work focuses on vaccines for enteric diseases such as cholera and typhoid, bacterial causes of meningitis and pneumonia and certain Flavivirus infections (dengue fever and Japanese encephalitis). The field research activities of the IVI are occurring in 22 countries of Asia, Africa, and Latin America.

IVI’s dual roles of research and capacity building expertise in topics throughout the vaccine continuum, including vaccine discovery, development, production, clinical evaluation, economic analysis, and policy research. This portfolio gives the institute a unique niche in the landscape of players working on new vaccines for developing countries.

The long term success of a venture is dependent both on the individuals leading the project and the quality of those delivering key functions. A real problem for the creation and sustainability of industry and consequently wealth in developing countries is the lack of individuals with the skills to generate initiatives and compete in the modern world. If you look at Africa in particular there is virtually no vaccine industry and very few trained individuals to start such initiative, possibly with the exception of South Africa. At the moment African Universities are struggling to compete academically and the flow of well trained individuals into science is limited. Even when good African scientists emerge the local scientific career structure is virtually non-existent.

Nevertheless, talented individuals with the appropriate training are emerging. Some are being trained in Western universities and some at well-founded sites such as the Wellcome Trust Clinical Units. The challenge now is to maintain such individuals in the African university system in the hope that they can spark entrepreneurial initiatives that benefit themselves, the local economy and healthcare in the region. This is clearly a longer term aim but one that is well worth pursuing. Indeed, both Asia and South America are already well down the road in this direction in the area of vaccinology as well as other areas such as diagnostics. The key to success in this area is long term investment and the gradual development of the areas of support as individuals move out of well funded environments involving a mixture of local and imported scientists into new laboratories of their own.

Conclusions I have presented a highly personalised view of the field of vaccinology and how it is developing at a global level. There are many encouraging signs that we are entering a new era where vaccines might get to all who need them. However, the task is still very much in hand and we as a community should find ways of making sure that these efforts are eventually successful.

acknowledgements â– The author is supported by the Wellcome Trust

Professor Gordon Dougan The Wellcome Trust Sanger Institute

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Tuberculosis vaccines — an update The current tuberculosis (TB) vaccine Mycobacterium bovis bacillus Calmette-GuÊrin (BCG) is the most widely used vaccine worldwide, but it does not prevent the establishment of latent TB or reactivation of pulmonary disease in adults. Peter Andersen looks at the progress of the candidates to improve or replace BCG This article was originally published in Nature Reviews Microbiology, July 2007, Vol 5, No. 7 p484-487 http://www.nature.com/reviews/micro doi:10.1038/nrmicro1703. It is reproduced with kind permissions of the Journal Editor, Author and Publisher

Estimated incidence rates per 100,000, all forms of tuberculosis 0-24 25-49 50-99 100-299 >300 No estimate

Figure 1. The distribution of tuberculosis in 2003 (data taken from WHO)

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uberculosis (TB), a disease which is both curable and preventable, still kills 2–3 million people every year. After decades of neglect, the immense public health impact of TB is now widely recognized, and the development of new tools to combat and control the epidemic has become an international priority. The current strategy for TB control is based on reducing the spread of infection through effective treatment of individuals with active disease and vaccination of children. The WHO has initiated the directly observed therapy (DOTS) campaign in many regions, but so far this programme has not been able to control the global TB epidemic or prevent the increase in multidrug resistant (MDR) strains of Mycobacterium tuberculosis (WHO. Global tuberculosis control — surveillance, planning, financing. [online] (2004).

The current TB vaccine Mycobacterium bovis bacillus Calmette–Guérin (BCG) is the most widely used vaccine worldwide. BCG provides efficient protection against TB in newborns, but does not prevent the establishment of latent TB or reactivation of pulmonary disease in adults. Being a viable organism, the activity of BCG depends on its initial replication, and it therefore cannot be used as a booster in an adult population that is already sensitized by prior BCG vaccination, exposure to environmental mycobacteria or latent TB (Anderson & Docherty, 2005). A novel, effective vaccination strategy against adult pulmonary TB is therefore a crucial goal and an active field of research, development and clinical evaluation.

Global distribution and disease burden In 2004, approximately nine million people developed active TB. Although this places TB as one of the most important global health problems, active disease represents only the tip of the iceberg, as it has been estimated that one-third of the world’s population is latently infected with M. tuberculosis. Globally, the incidence of TB is growing, mainly owing to the spread of HIV in Africa, where it has been estimated that 13% of adults with newly diagnosed TB are also coinfected with HIV (Dye, 2006).

However, in recent years, the increasing TB problem in Eastern European countries has contributed to the worsening global epidemic. Africa has the highest estimated incidence (356 per 100,000 population per year), but major parts of Asia also have a significant TB problem (WHO. Global tuberculosis control — surveillance, planning, financing. [online] (2004) Figure. 1. In most of these regions, the incidence of TB has now reached such a magnitude that it is overwhelming the limited resources available to identify and treat active contagious pulmonary TB. Furthermore, by primarily targeting

induced during natural M. tuberculosis infection, and although they do not seem to have a major role in the initial control of the infection, they might be more involved in the later, chronic stages of the disease (Woodworth & Behar, 2006). To target this lymphocyte subset, some of the new vaccines are delivered through live carriers such as viral vectors or genetically modified strains of BCG. In the 5–10% of latently infected individuals who go on to develop active TB, the balance between the natural immunity of the host and the pathogen is thought to change, for example, following an

Table 1. The leading tuberculosis vaccine candidates in clinical trials Vaccine name

Vaccine type

Development stage

Institute or company

rBCG30

Live, recombinant BCG

Clinical Phase I completed in 2004

UCLA School of Medicine/ Aeras

rBCG ∆ureC:HLy

Live, recombinant BCG

Clinical Phase I planned for 2007

Max Planck Institute for Infection Biology/Vakzine Projekt Management

MVA-85A

Modified vaccinia virus

Completed and ongoing Clinical Phase I

University of Oxford

H1/IC31

Adjuvanted subunit

Completed and ongoing Clinical Phase I

Statens Serum Institute/ Intercell

Mtb72f/AS02A

Adjuvanted subunit

Completed and ongoing Clinical Phase I

GlaxoSmithKline

H1/LTK63

Adjuvanted subunit

Clinical Phase I ongoing

Statens Serum Institute/ Novartis

HyVac4 /IC31

Adjuvanted subunit

Clinical Phase I planned for mid-2007

Statens Serum Institute/ Intercell/ Aeras

BCG = Mycobacterium bovis bacillus Calmette-Guérin

the working population, TB is a major roadblock to healthy economic development in many developing countries.

Immunity to M. tuberculosis M. tuberculosis infection remains latent with no overt clinical symptoms throughout life in more than 90% of infected individuals. Progressive mycobacterial infection in patients with deficient interferon-γ (IFN-γ) and tumour necrosis factor (TNF) signalling provides convincing evidence for the importance of these cytokines in the control of TB. The major source of these cytokines are CD4+ T cells, the most important lymphocyte population in the protective immune response and the main target for most vaccination strategies (Kaufmann, 2005). The role of CD8+ T cells is less clear. They are

immunosuppressive event, resulting in massive bacterial replication and reactivation of the disease.

All of the new TB vaccine candidates that are under clinical evaluation (Table 1) are designed as pre-exposure vaccines and, hence, are aimed at stimulating an immune response that controls subsequent infection more efficaciously than the immune response that is stimulated during natural infection, thereby delaying reactivation.

It is not known whether post-exposure administration of these vaccines to already latently infected individuals would prolong host containment of latent TB and prevent reactivation, or whether this would require specially designed post-exposure vaccines based on antigens that are expressed by the bacteria during latency, as recently discussed elsewhere (Andersen, 2007).

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Vaccine concepts and clinical trials Current attempts to develop improved TB vaccination strategies can be divided into two approaches — replacing or boosting BCG. The first strategy aims to replace BCG with a more effective vaccine. This is generally believed to demand an improved, attenuated mycobacterial vaccine strain, obtained either through the generation of gene-deletion mutants of M. tuberculosis, or by re-introducing important antigens or other factors into the existing BCG vaccine strain. Viable, attenuated mycobacterial vaccines obviously present a broad variety of antigens and will potentially cover a combination of different T-cell populations, but such vaccines must be not only more potent than BCG, but also at least as safe, in order to be considered as candidates for clinical trials (Kamath et al., 2005).

The second strategy involves the development of a booster vaccine that takes advantage of BCG priming vaccination in childhood, and is given to increase the immune response and prolong immunity to also cover the adult population. It is generally agreed that such a vaccination strategy can be best accomplished with a subunit vaccine. Subunit vaccines are based on a restricted number of antigens and hence on a highly focused immune response for protection. In several of the leading vaccine candidates, the individual antigens are fused into polyproteins, something that both increases the immunogenicity of the individual antigens and has obvious advantages from a manufacturing point of view. The success of the booster strategy is underpinned by recent advances in adjuvant development.

Until recently, the only adjuvants appropriate for use in TB vaccines were either ineffective at stimulating T-cell responses or were too toxic for human use. This situation has rapidly changed in recent years, and a number of novel, promising T-cell adjuvants such as the

IC31 adjuvant, cationic liposomes, the AS2 formulation and LTK63 (for mucosal delivery) are now under latepreclinical or clinical development in TB vaccines (Table 1). Eventually, the ultimate vaccine strategy could be based on a combination of both approaches, that is, a prime–boost vaccination regime that comprises priming with the best possible viable vaccine candidate and boosting with the best possible subunit vaccine candidate (Kaufmann, 2005).

BCG replacement vaccines rBCG30 is a recombinant BCG vaccine in which the well-known and well-characterized antigen 85B (Ag85B) is overexpressed. This 30 kDa enzyme, which is involved in outer cell-wall synthesis, is a key component in several TB vaccines, and although Ag85B is already abundantly secreted by BCG, overexpression appears to increase responses to this important antigen (Horwitz & Harth 2003). rBCG30 has been tested in a Phase I trial in humans and was well tolerated. rBCG ∆reC:Hly. To amplify the CD8+ T-cell response induced by BCG, a recombinant BCG mutant has been constructed that expresses listeriolysin (Hly), which can perforate the phagosome membrane. The gene (ureC) encoding the urease enzyme that increases the pH of the phagosome containing BCG was additionally deleted to avoid neutralizing the phagosome, as this would reduce the activity of Hly (Grode et al., 2005).

Surprisingly, apoptosis of infected macrophages and cross-priming of dendritic cells seems to be the major mechanisms responsible for the increased activity of this vaccine (Winau et al., 2006). A clinical Phase I trial is planned to commence by the end of 2007.

BCG booster vaccines The Ag85B–ESAT6 fusion molecule (H1) is made up of the two secreted antigens Ag85B and ESAT6. These individual antigens both have an

Box 1. Key areas for tuberculosis (TB) vaccine development ■ Determine the correlates of vaccine induced protective immunity ■ Determine the requirements for post-exposure TB vaccines ■ Understand vaccine-induced T-cell memory to Mycobacterium tuberculosis ■ Determine the role of CD8+ T cells in M. tuberculosis infection and immunity

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impressive track record of studies confirming their antigenicity in humans and their vaccine potential. H1 has shown promise both for parenteral (in IC31 or cationic liposomes) and mucosal (in LTK63) delivery (Agger et al., 2006; Dietrich et al., 2006). In addition to being a valuable vaccine component, ESAT6 (the component of H1 localized in the region that was deleted during the original attenuation of BCG, and which is therefore absent from all BCG vaccine strains) is a key component in a new generation of diagnostic tests for M. tuberculosis infection (Pai, Riley & Colford 2004).

An alternative fusion construct, called H4, has been engineered and consists of Ag85B and the TB10.4 antigen, which is also from the ESAT family of small secreted antigens (Dietrich et al., 2006). TB10.4 has similar immunological properties to ESAT6, but it is highly expressed and immunodominant in BCG. H4 is a powerful booster vaccine for BCG, whereas the H1 vaccine for comparison, in addition to boosting Ag85B responses, will supplement the BCG antigen repertoire with the important ESAT6 antigen component.

H1 is currently in clinical trials administered both parenterally and through the mucosal route. The first clinical trial in Leiden, Holland (Dissel & Ottenhoff, unpublished data) evaluated the vaccine in a conventional parenteral vaccination strategy, using the IC31 adjuvant. This trial was conducted in purified protein derivative (PPD)-negative individuals and the vaccine was shown to be both safe and strongly immunogenic. The H1/IC31 vaccine is currently being evaluated in PPD-positive BCG-vaccinated individuals at the same site. Another trial that has recently started will test the nasal administration of the H1 antigen, using the LTK63 adjuvant. The H4/IC31 vaccine will commence clinical trials in mid-2007 in Sweden.

The MTB72f vaccine is a fusion molecule consisting of two antigens that are strong targets for T helper 1 (TH1) cells in PPD-positive individuals.

Rv1196 (MTB32) is inserted into the middle of the serine protease Rv0125 (MTB39), which is thus present as two fragments (Sheiky, et al., 2004).

MTB72F in the AS02A adjuvant formulation has recently completed two Phase I trials in healthy PPD-negative

adults in the United States and Belgium.

The vaccine was well tolerated and safe, and could induce both antigen-specific humoral and cell-mediated immune responses.

MVA85A is a modified vaccine virus Ankara (MVA) strain expressing antigen 85A, another member of the Ag85 family of protective antigens. In Phase I studies in humans, MVA85A was found to be safe and well tolerated, and this vaccine has induced strong immune responses, particularly in previously BCG-vaccinated individuals16.

Conclusions With increasing investment from public funds such as the European Union, National Institutes of Health and the Bill & Melinda Gates Foundation in recent years, TB vaccine research, development and evaluation has become an active area, with several vaccines in various stages of early clinical development. Most of this work is conducted by public research organizations and public–private partnerships, but a recent re-analysis and demonstration of the significant commercial value of a novel TB vaccine (BIO Ventures for Global Health, 2006 [online]) will probably result in a larger investment from private industry. This will promote streamlined development and the eventual global distribution of a novel vaccine. Although a new, improved vaccination strategy against TB is finally on the horizon, its eventual success will still depend on continued close integration with information from basic research. The identification of reliable correlates of protection, as well as the answers to more basic immunological questions relating to immunological memory and the relative importance of different T-cell subsets, will be important for the potential modification of the leading TB vaccines, the generation of secondgeneration products and the selection of which vaccines to move forward into expensive efficacy trials (Box 1). It will furthermore be a high priority for the clinical development programmes to evaluate whether the current vaccines, all of which have been designed for preinfection administration, will also prevent reactivation of TB if administered post-exposure to the large proportion of the global population already latently infected with TB.

references ■ Agger, E. M. et al. Protective immunity to tuberculosis with Ag85B-ESAT-6 in a synthetic cationic adjuvant system IC31. Vaccine 24, 5452–5460 (2006).

■ Andersen, P. & Doherty, T. M. The success and failure of BCG — implications for a novel tuberculosis vaccine. Nature Rev. Microbiol. 3, 656–662 (2005).

■ Andersen, P. Vaccine strategies against latent tuberculosis infection. Trends Microbiol. 15, 7–13 (2007).

■ BIO Ventures for Global Health. Tuberculosis vaccines: The case for investment [online] (2006).

■ Dietrich, J. et al. Exchanging ESAT6 with TB10.4 in an Ag85B fusion molecule-based tuberculosis subunit vaccine: efficient protection and ESAT6-based sensitive monitoring of vaccine efficacy. J. Immunol. 174, 6332–6339 (2005).

■ Dietrich, J. et al. Mucosal administration of Ag85B-ESAT-6 protects against infection with Mycobacterium tuberculosis and boosts prior Bacillus Calmette–Guerin immunity. J. Immunol. 177, 6353–6360 (2006).

■ Dye, C. Global epidemiology of tuberculosis. Lancet 367, 938–940 (2006).

■ Grode, L. et al. Increased vaccine efficacy against tuberculosis of recombinant Mycobacterium bovis bacille Calmette–Guerin mutants that secrete listeriolysin. J. Clin. Invest. 115, 2472–2479 (2005).

■ Horwitz, M. A. & Harth, G. A new vaccine against tuberculosis affords greater survival after challenge than the current vaccine in the guinea pig model of pulmonary tuberculosis. Infect.

Immun. 71, 1672–1679 (2003).

■ Kamath, A. T. et al. New live mycobacterial vaccines: the Geneva consensus on essential steps towards clinical development. Vaccine 23, 3753–3761 (2005).

■ Kaufmann, S. H. Recent findings in immunology give tuberculosis vaccines a new boost. Trends Immunol. 26, 660–667 (2005).

■ McShane, H. et al. Recombinant modified vaccinia virus Ankara expressing antigen 85A boosts BCG-primed and naturally acquired antimycobacterial immunity in humans. Nature Med. 10, 1240–1244 (2004).

■ Pai, M., Riley, L. W. & Colford, J. M. Jr. Interferon— assays in the immunodiagnosis of tuberculosis: a systematic review. Lancet Infect. Dis. 4, 761–776 (2004).

■ Skeiky, Y. A. et al. Differential immune responses and protective efficacy induced by components of a tuberculosis polyprotein vaccine, Mtb72F, delivered as naked DNA or recombinant protein. J. Immunol. 172, 7618–7628 (2004).

■ Winau, F. et al. Apoptotic vesicles crossprime CD8 T cells and protect against tuberculosis.

Immunity 24, 105–117 (2006).

■ Woodworth, J. S. & Behar, S. M. Mycobacterium tuberculosis-specific CD8+ T cells and their role in immunity. Crit. Rev. Immunol. 26, 317–352 (2006).

■ WHO. Global tuberculosis control — surveillance, planning, financing. [online] (2004).

acknowledgements ■ Vaccine research in the Andersen laboratory is supported by EU-FP6 TBVAC, EU-FP6 MUVAPRED, grants from the Danish Research Council and the Bill & Melinda Gates Foundation, Grand Challenges in Global Health (GC6, GC12) and the AERAS Global Vaccine Foundation.

further information ■ Bill & Melinda Gates Foundation: http://www.gatesfoundation.org/default.htm ■ WHO Stop TB: http://www.who.int/tb/en

Peter Andersen, Statens Serum Institute, Denmark

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Revealing all: the microscopic world of compost at the Chelsea Flower Show Linda Thomas and Graham Kinsey wowed the crowds with a fascinating glimpse into the world of compost

C

ontinuing the great tradition of microbiology at the Chelsea Flower Show, an opportunity was created for us to present an educational exhibit to the public, working with the Hillier’s exhibit ‘Planting for Trees’ in the Great Pavilion, sponsored by Dr Julian Kenyon’s Dove Clinic for Integrated Medicine (www.doveclinic.com). Yakult and Germains Seed Technology (www.germains.com) contributed to this sponsorship by providing a live view into the microbial world of compost, which was a good contrast to the rather more macroscopic biology of the trees that formed the bulk of the exhibit.

This was the first time the Royal Horticultural Society (RHS) had allowed an educational exhibit to form part of such a display. A light microscope, housed in a small summerhouse at the edge of the garden, was used to examine samples of home-made compost, with the images displayed directly on a monitor on the outside wall. We are both microbiologists working in companies interested in the benefits and use of beneficial microorganisms in different ways. Germains Seed Technology develops seed coating and technology products using beneficial microorganisms to coat seeds. This helps to exclude pathogens and give a competitive advantage to the young seedlings. Of course, probiotics such as Yakult work in a similar way in the gut — by the phenomenon of colonisation resistance — competing against harmful microorganisms for nutrients and space.

The week was great fun, albeit exhausting and not always comfortable. Conditions fluctuated from miserably cold to boiling hot in the rather too cosy summerhouse, but there was a great sense of achievement in bringing such a display to the public. One of us would work inside on the microscope hunting for interesting and moving microorganisms, whilst the other explained the images to the interested public outside. The largest organisms were nematodes; the smallest were the bacteria. Mostly we saw a range of protozoa; ciliates and flagellates, moving around on the screen. The biggest excitement was when we saw, live, a protozoa dividing into 4 offspring.

Most people were completely fascinated — only a few shivered in disgust. Our aim was to explain and promote the benefits of making your own compost, which in turn helps increase the numbers of beneficial microorganisms in the garden soil. This benefits the plants by recycling and making nutrients more available to them, keeping a better balance of microbes in the soil and helping to reduce plant diseases

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The team: (Left to Right): Dr Linda Thomas (Yakult UK Ltd), Andy McIndoe with Gold Medal (Hiller Garden Centre), Dr Graham Kinsey and Jo Howell (Germains Seed Technology)

...and all for free! We handed out thousands of educational leaflets, and talked endlessly to many interested gardeners, fascinated by this microbial world of compost revealed to them for the first time. It was a great delight to show the microscope and talk to the few children that were there; I hope we may have inspired some budding young scientists.

We felt great joy (and relief) on Tuesday morning when we heard that Hillier had won their 62nd consecutive Gold Medal. It was well deserved for a really marvellous display of trees and shrubs. All in all, this was a unique and worthwhile experience. We hope we inspired many more compost makers and opened the eyes of gardeners to the value of beneficial microbes.

Dr. Linda Thomas Yakult UK Ltd

Dr. Graham Kinsey Germain’s Technology, UK

Stat Note 10 Anthony Hilton

In the tenth of a series of articles about statistics for biologists, Anthony Hilton and Richard Armstrong discuss: Richard Armstrong

The two-way analysis of variance

I

n a previous article in Microbiologist, Vol 8, No.2 July 2007 (Armstrong & Hilton, 2004), we described a one-way analysis of variance (1-way ANOVA) in a randomised design. In a 1-way ANOVA, an individual observation is classified according to which group or treatment it belongs and observations within each group are a random sample of the relevant population. The scenario to illustrate this analysis compared the degree of bacterial contamination on 2p coins collected from three types of premises, viz., a butcher’s shop, a sandwich shop, and a newsagent. A sample of four coins was collected at random from each location and the number of bacterial colonies present on each coin was estimated.

Such an experiment is often described as in a ‘randomised design’. More complex experimental designs are possible, however, in which an observation may be classified in two or more ways (Snedecor & Cochran, 1980).

The scenario We return to the scenario described in Statnote 7 (Hilton & Armstrong, 2006a). A hypothetical experiment was carried out to investigate the efficacy of two novel media supplements (S1 and S2) in promoting the development of cell biomass. Three 10-litre fermentation vessels were sterilised and filled with identical growth media with the exception that the media in two of the vessels was supplemented with 10 ml of either medium supplement S1 or S2. The vessels were allowed to equilibrate and were subject to identical environmental / incubation conditions. The vessels were then inoculated with a culture of Bacterium x at an equal culture density and the fermentation allowed to proceed until all the available nutrients had been exhausted and bacterial growth had ceased. The entire volume of culture media in each fermentation vessel was then removed and filtered to recover the bacterial biomass, which was subsequently dried and the dry weight of cells measured.

This experiment was originally carried out with 25 replicate vessels of each treatment and were analysed using a 1-way ANOVA in a randomised design; i.e., the treatments were allocated at random and without restriction to the original vessels (Hilton & Armstrong, 2006a). The present experiment, however, was carried out in a different way. First, 30 vessels were divided into 10 groups of three, each group representing ‘a replication’ with the intention of setting up and processing each replication (a control and each of the two treatments S1, S2) on each of 10 separate occasions and second, the treatments were allocated to the three vessels within a replication independently and at random.

The analysis appropriate to this design is analogous to that of the paired sample ‘t-test’ described in Statnote 3 (Hilton & Armstrong, 2005) but extended to more than two treatments.

Terminology In a two-way design, each treatment is allocated by randomization to one experimental unit within each group.

The name given to each group varies with the type of experiment. Originally the terminology ‘randomized blocks’ was applied to this type of design because it was first used in agricultural experiments in which experimental treatments were given to units within ‘blocks’ of land, plots within a block tending to respond more similarly compared with plots in different blocks (Snedecor & Cochran, 1980). In the present example, the block is a single trial or replication of the comparison between treatments, the trial being carried out on 10 separate occasions. Furthermore, in experiments with human subjects, there is often considerable variation from one individual to another and hence a good strategy is to give all treatments successively to each ‘subject’ in a random order; the subject therefore comprising the ‘block’ or ‘replication’.

Table 1. The effect of two novel media supplements (S1, S2) on bacterial biomass measured on 10 separate occasions Occasion (‘blocks’)

Control

+S1

+S2

1

461

562

344

2

472

573

359

3

473

574

369

4

481

581

403

5

482

582

425

6

494

586

476

7

493

591

511

8

495

592

513

9

506

592

556

10

502

607

578

The statistical model In the example given in Table 1, the fact that vessels are also grouped into replications, one complete replication for each of the 10 occasions, gives a more complex model. Using the commonly used notation to describe the basic model of an ANOVA described in Statnote 9 (Hilton & Armstrong, 2007a), September 2007

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features

the two-way design includes a term for the replication effect ‘b’ in addition to the treatment effect ‘a’, viz., xij = µ + ai + bj + eij Hence, the ANOVA table (Table 2) includes an extra term for replications, i.e., the occasion on which the replication was sampled. In the terminology used in Statnote 9 (Hilton & Armstrong, 2007a), treatment (ai) is a fixed-effect factor whereas blocks or occasions (bj) are a ‘random-effect’ factor.

In addition to the assumptions made in the randomized design, viz., homogeneity of variance, additive class effects, and normal distribution of errors (Armstrong et al., 2000), this type of design makes the additional assumption that the difference between treatments is consistent across all replications (Snedecor & Cochran, 1980).

Table 2. Analysis of variance (two-way in randomised blocks) of the data in Table 1 ANOVA table: Variation

SS

DF

MS

F

Treatments

91373.4

2

45686.7

27.32*

Replications

35896.0

9

3988.45

2.39

Error

30097.3

18

1672.07

* P < 0.001, + Not quite significant at P = 0.05; Post-hoc tests (Scheffé): Control v.

S1 S = 14.39*, Control v. S2 S = 1.48 (P > 0.05), S1 v. S2 S = 25.11*

Conclusion The two-way design has been variously described as a matched-sample F-test, a simple within-subjects ANOVA, a one-way within-groups ANOVA, a simple correlated-groups ANOVA, and a one-factor repeated measures design! This confusion of terminology is likely to lead to problems in correctly identifying this analysis within commercially available software. The essential feature of the design is that each treatment is allocated by randomization to one experimental unit within each group or block. The block may be a plot of land, a single occasion in which the experiment was performed, or a human subject. The ‘blocking’ is designed to remove an aspect of the error variation and increase the ‘power’ of the experiment. If there is no significant source of variation associated with the ‘blocking’ then there is a disadvantage to the two-way design because there is a reduction in the DF of the error term compared with a fully randomised design thus reducing the ‘power’ of the analysis.

Dr Anthony* Hilton and Dr Richard Armstrong** *Pharmaceutical Sciences and **Vision Sciences, Aston University, Birmingham, UK

Interpretation of the results The ANOVA appropriate to the two-way design is shown in Table 2. The design is often used to remove the effect of a particular source of variation from the analysis. For example, if there was significant variation due to replications and, if treatments had been allocated to vessels at random, then all of the ‘between occasions’ variation would have been included in the pooled error variance. The effect of this would be to increase the error variance and to reduce the ‘power’ of the experiment (Hilton & Armstrong, 2007b) thus making it more difficult to demonstrate a possible treatment effect. In a two-way design, however, variation between replications, attributable to occasions, is calculated as a separate effect and therefore, does not appear in the error variance. This may increase the ‘power’ of the experiment and make it more probable that a treatment effect would be demonstrated. In the example quoted (Table 2), there is a highly significant effect of treatment (F = 27.32, P < 0.001). In a two-way design, planned comparisons between the means or post-hoc tests can be performed as for the randomized design (Hilton & Armstrong, 2006b). Hence, Scheffé’s post-hoc test (Hilton & Armstrong, 2006b) suggested that this result is largely due to the effect of supplement S1 increasing yield. In addition, in a two-way design, the variation due to ‘replications’ is calculated (F = 2.39) and this was not quite significant at P = 0.05. The borderline significance suggests there may have been some differences between replications and removing this source of variation from a comparison of the treatment effect may have increased the power of the experiment.

A comparison of the ANOVA table in Table 1 with that for a one-way ANOVA in a randomised design demonstrates that reducing the error variance by ‘blocking’ has a cost, viz., a reduction in the degrees of freedom (DF) of the error 42

variance which makes the estimate of the error variation less reliable. Hence, an experiment in a two-way design would only be effective if the ‘blocking’ by occasion or some other factor reduced the pooled error variance sufficiently to counter the reduction in DF (Cochran & Cox, 1957; Snedecor & Cochran, 1980).

September 2007

references ■ Armstrong R A, Slade S V & Eperjesi F (2000) An introduction to analysis of variance (ANOVA) with special reference to data from clinical experiments in optometry. Ophthal Physiol Opt 20: 235-241.

■ Armstrong R A & Hilton A (2004) The use of analysis of variance (ANOVA) in applied microbiology. Microbiologist Vol 5: No. 4 Dec 2004, pp 18 - 21.

■ Cochran, W.G. & Cox, G.M. (1957) Experimental designs. Second Ed., John Wiley, New York, London and Sydney.

■ Hilton A & Armstrong R A (2005) Statnote 3: Testing the difference between two groups. Microbiologist Vol 6: No.4 Dec 2005, pp 30-32.

■ Hilton A & Armstrong R A (2006a) Statnote 5: Is one set of data more variable than another? Microbiologist Vol 7: No.2 June 2006, pp 34-36.

■ Hilton A & Armstrong R A (2006b) Statnote 6: Post-hoc ANOVA tests. Microbiologist Vol 7 No.3 Sep 2006, pp 34-36.

■ Hilton A & Armstrong R A (2007a) Statnote 9: The one-way analysis of variance (random effects model): the ‘nested’ or ‘hierarchical’ design. Microbiologist Vol 8: No.2 June 2007, pp 4041.

■ Hilton A & Armstrong R A (2007b) Statnote 8: Statistical power and sample size. Microbiologist Vol1 8: No.1 March 2007, pp 38-40.

■ Snedecor G W & Cochran W G (1980) Statistical methods. 7th edition, Iowa State University Press, Ames, Iowa.

members

careers

Andrew Traube describes the diverse responsibilities and exciting opportunities offered by a career in Clinical Research

Clinical Research and the Clinical Research Associate

M

y first real insight into microbiology came during my A-Levels and was furthered during my degree: â&#x20AC;&#x2DC;Applied and Human Biology,â&#x20AC;&#x2122; which I undertook at Aston University.

This was fundamentally a diverse course covering a wide range of topics such as biodiversity, immunology and physiology, as well as virology, mycology and microbiology. The course also gave me the opportunity to undertake an industrial placement during the third year of study. This placement year was spent at the Queen Elizabeth Hospital in Birmingham, where I undertook the equivalent of the first year of training required to become a medical laboratory scientific officer 1 (MLSO1). This sparked my interest to pursue a career in a

microbiology-related field, hence my final year options were tailored accordingly. That year, aside from the statutory exams, culminated in the undertaking of a research project, through which I wanted to utilise the skills and knowledge I had obtained during my placement year, coupled with some molecular biology or genomics. I felt this would stand me in good stead for a future career in microbiology.

For 10 weeks in the twilight of my degree, I studied the formation of triplex DNA within bacterial cells and tried to harness this potential to serve as an effective antibacterial agent.

Further funding meant the extension of the project for an additional three years, with my participation upon completion of my degree, to form my Ph.D. The reason for undertaking a

Ph.D. was not solely to extend my experience as a microbiologist and to develop my laboratory skills. I saw it as means to expand my so-called softer skills such as communication, project management and organisational skills; skills that would be invaluable in my current role. At the time, I believed this would be a necessity in pushing my career down the research route.

After completing my Ph.D., I felt a move into industry would serve me well. Having gained experience in academic and clinical laboratories, this move seemed the next logical step, not only in providing another aspect of research experience, but also in gaining research experience driven by commercial forces and pressures.

My Ph.D. led me into a job with Depuy CMW, which is part of the Depuy

September 2007

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and the fact that it enabled me to utilise my microbiological and other scientific learning, without being laboratory bound. It is also at the forefront of drug development, providing insight into the drugs of the future.

As a clinical monitor you can be responsible for overseeing trials at any of the four phases of drug development (Figure 1). As a result you can be involved in trials looking simply at the tolerability and pharmacokinetics of drugs in healthy humans or in drugs that have received marketing approval, along with anything in between.

There are generally two routes people take as a CRA — either people join a contract research organisation (CRO) or they join a pharmaceutical company. These are not mutually exclusive; and a lot of CRAs aim to gain experience in both, but tend to eventually settle in one or the other depending on their personal preference.

International group, the orthopaedic subsidiary of Johnson & Johnson.

Utilising the analytical techniques and knowledge I had gained during my Ph.D., I secured a job as an analytical scientist working on a project looking at the release characteristics of antibacterial agents. It was towards the end of my Ph.D. and around this time that I began looking at alternative careers that would involve a move away from the laboratory. I was looking for a career with a more structured career path, one that had definite goals and clear, definable promotional steps. In my search for this ‘dream job,’ I came across clinical research and the role of clinical monitors or Clinical Research Associates (CRA).

Figure 1. The phases of a clinical trial Phase 1 Sets out to establish tolerability, pharmacokinetics and pharmacodynamics of a given drug, usually in young healthy volunteers.

The CRO

Phase 2 'First in to patient' trials in which patients inflicted with the disease the therapy has been designed to act upon, are administered the trial drug.

Phase 3 Pivotal large-scale trials using patients inflicted by the disease the drug is meant to act upon. Data generated is often used for Marketing Authorisation Application (MAA), the granting of which is necessary for the general use of the drug.

Phase 3b Non-pivotal trials started prior to launch of the drug, where usually the market leader is used as the comparator drug to establish possible benefits of the study drug, in an attempt to bolster market share once the MAA is granted.

Phase 4 Post-authorisation studies used to capture further drug data, including safety information, once the drug has been released to the general population.

The CRA Role The primary role of the CRA is to ensure drug trials are run ethically from the patient’s perspective, and that the sites they monitor comply with all regulatory and ethical requirements.

Because drug trials are performed on human subjects in varying disease states, the field is very heavily regulated. The role itself is basically a project management position in which the CRA oversees a number of hospital sites or clinics at which the test drug/treatment is being trialed, ensuring the study is performed correctly and according to protocol.

The appeal of the role was its variability 44

Working for a CRO is variable and generally offers studies in a range of therapeutic areas. The trials taken on

by CROs come from pharmaceutical and biotechnology companies, termed the ‘sponsor,’ that outsource the work.

Anything from small start-ups to large, world leading, multinational pharmaceutical companies outsource drug trials to CROs; hence it is clear why the work is so varied. A clinical trial is made up of a whole host of elements, and a CRO is equipped with the knowledge and wherewithal to undertake either a single aspect of the trial, such as data management, or conduct the full development.

It is generally the case that a CRA working in a CRO will be assigned two to three studies, all of which can be in completely separate therapeutic areas.

It is unusual for the CRA to remain on a study through to completion, and there is more freedom to move from study to study within this environment.

Experience-wise, working for a CRO enables a CRA to gain a broader spectrum of expertise, which is particularly important during the early stages of a career as a CRA.

Pharmaceutical Companies CRAs in the pharmaceutical company will tend to be allocated to a single study drug and are more likely to monitor that trial to its conclusion.

September 2007

Here the CRA will have a greater involvement with the study drug and scientific knowledge/background may be more heavily drawn upon. One word of warning: whether you undertake the role in a CRO or a pharmaceutical company, life as a CRA generally involve a lot of travel, as sites allocated to a study are selected on the basis of the investigator speciality and their expertise in running a trial in a specific therapeutic area. As a result, a CRA assigned to a study can have sites at the opposite ends of the country. If travel is not something that appeals or your home life is not receptive to this, then this is not the career for you.

Clinical Research Clinical research is a very popular area, particularly the role of a CRA, and as a result it is a very competitive market. The hardest move is the initial entry point, which can be quite problematic, something I found quite difficult. The problem is that all lower end positions, those that will give you the necessary experience to become a CRA, will still normally require some form of prior clinical experience. This can come in the form of a nursing background, previous experience of clinical trial process through CTA (clinical trial administrator/assistant), CDA (clinical data assistant) or clinical safety scientist roles. A study coordinator role will also stand applicants in good stead, as such roles will provide experience in the running of the clinical trial from the hospital perspective, rather than from the pharmaceutical/CRO side. As a result, these positions have direct experience of the trial process but from a site point of view. Without any experience in clinical trials, it is very difficult to gain access to the field and nigh on impossible to obtain a role as a CRA. My own personal entry to the field was slightly different, as I did not have any specific clinical trial experience. I had clinical experience in terms of working in a clinical laboratory, but whether or not this went in my favour is not entirely clear, as it provided no clinical trial experience. My Ph.D. coupled with the experience gained working for Depuy, which itself was heavily regulated, working strictly to GMP (good manufacturing practice) and GLP (good laboratory practice), along with an understanding of the

clinical trial process gained from personal research, enabled me to gain entry to the industry. However, I will say that in order to fulfil my ambition to become a CRA, and due to my lack of clinical experience, I have had to take a step backward, both financially and in terms of career progression, in the short term. This is not an uncommon phenomenon in the field, and a number of my work colleagues have done exactly that in order to obtain the same goal.

Figure 2. The possible paths of promotion for a Research Assistant RA

▼ CRA 1

▼ CRA 2

▼ Senior CRA 1

A day in the Life of an RA

▼ ▼

Senior CRA 2

▼ ▼

▼ ▼

Principle CRA

▼

Assistant COM

▼ Associate COM

▼

CTM

▼

▼ COM

▼

PM

KEY RA:

Research Assistant

CRA:

Clinical Research Associate

CTM:

Clinical Team Manager

PM:

Project Manager

COM:

Clinical Operations Manager

▼ AD COM

▼ D COM

AD COM: Assistant Director COM D COM:

their head European office, based in Cambridge, U.K. The training with PPD usually takes a year, which culminates in internal and external assessments that have to be successfully passed in order to be promoted to a CRA. During the year as an RA, you are assigned to anything from one to six studies (usually three to four), where you will support CRAs, clinical team managers (CTMs) and project managers (PMs) providing you with direct experience and knowledge of the industry and the clinical trial process. This is enhanced with co-monitoring visits, where the CRAs provide on-site training to the RA in aspects of monitoring, such as source data verification and query solving. The RA is allowed to sit in on discussions with the investigators and pharmacy, all tasks that will form part of their own visits in the future. These visits serve to provide specific on-thejob training and give a full insight into what monitoring entails.

Director COM

That said, I believe that becoming a CRA will make this all worthwhile.

Working as a CRA provides a very structured career path enabling relatively quick promotion for successful candidates, with numerous career options/paths available, some of which are illustrated in Figure 2. Also, training as a CRA does not just restrict you to a career involved with the running of clinical trials; moves from CRA to regulatory affairs and medical writing, to name just two, are not uncommon.

My Current Position I am currently (at the time of going to press) seven months into my year as a research assistant (RA) with PPD at

A day in the life of an RA is difficult to define as the job is so variable and, to a degree, you have to be quite reactive/adaptive. I can walk into the office at 9:00 a.m. with a list of jobs to cover that day, and then walk out at 5:30 p.m. having not finished one of those tasks because tasks with a higher priority have arisen. Issues involving ethics/regulatory submissions, collection of specific information from worldwide CRAs requested by your CTM (sometimes very problematic due to the different worldwide time zones, particularly when dealing with CRAs in Australia or New Zealand) or sponsor audits are fast approaching. These are all things I have witnessed personally that have thrown my workload offline.

However, general day-to-day tasks of an RA will include minute-taking, document tracking and maintaining the trial master file (TMF) and/or the country files. Each trial has a master file that is maintained in accordance with company policy, which in turn will comply with regulatory and ethical standards and legislation. The TMF will detail a whole host of information from study start-up through to completion and will be added to right up until the study is closed. Country files follow a similar structure to the TMF but contain information specific to that country. This is due to regulatory and

ethical guidelines/legislation varying from country to country.

Document tracking can take up quite a lot of an RA’s time, as there is so much documentation that is required throughout the conduct of a clinical trial. It is worth bearing in mind here that this documentation is required on every patient at every site in every country involved in the trial.

Other tasks include essential document collection; corresponding with sites, CRAs, CTMs and PMs; involvement with the organisation of investigator meetings; and training sessions. The specifics of each RA’s role will vary in accordance with the studies they are assigned and the demands of the CRA, CTM or PM they support, as well as those of the sponsor company.

While some of these tasks appear somewhat monotonous, one of the downsides to the RA role, they provide essential knowledge and understanding of the structure of the clinical trial process. On the plus side, these tasks will only be performed for a year to 14 months after which “CRA-dom” beckons with the reward of gaining full responsibilities for your own sites along with all the monitoring tasks that come with it.

For anyone looking to start a career in clinical research, my advice is to do your research - the more you know about the industry, the better the chance you have of getting in. A good initial point of contact is the website for the Institute of Clinical Research (ICR www.instituteofclinicalresearch.org).

This is a very useful source of information and provides a whole of host of matter. Books, booklets and leaflets are available detailing the different aspects of clinical trials/research, as well as different job roles within the industry. The ICR runs seminars, workshops and conferences and even provides a qualification in clinical research that is recognised industry-wide. A lot of information is available to non-members, but for those serious about a move into clinical research, membership to the ICR is inexpensive and recommended, and entitles you to a monthly magazine containing fundamental information about industry-wide issues.

September 2007

Andrew Traube Research Assistant, PPD

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members

Key Stage 3 1st Prize winning entry

GCSE 1st Prize winning entry

Key Stage 3 2nd Prize winning entry

GCSE 2nd Prize winning entry

Key Stage 3 3rd Prize winning entry

GCSE 3rd Prize winning entry

46

September 2007

MiSAC Competition 2007 John Grainger reports on the Society’s special sponsorship of the 19th Annual MiSAC competition: Salmonella: from farm to fork

MiSAC is the Microbiology in Schools Advisory Committee. Its members represent a wide range of educational institutions and scientific organisations.

For further information visit: www.microbiologyonline.org.uk/what.htm

In addition to being a regular sponsor of Microbiology in Schools Advisory Committee (MiSAC), SfAM provided the special sponsorship for the 19th annual MiSAC competition Salmonella: from farm to fork. The topic was chosen to raise awareness of the serious health problems caused by Salmonella food poisoning.

The task of this years’ competition was to produce a leaflet for the general public, which arose from the need to raise consumer awareness of how such problems occur and how to avoid them. This year’s competition proved a very popular one. It generated nearly 950 entries, a near record, and involved more than 1000 students from 130 schools and colleges drawn from England, Wales, Scotland and Northern Ireland. Of the two age groups for the competition, Key Stage 3 provided more entries than GCSE. However, this year it was very pleasing to see a much larger number of entries from GCSE students than previously.

The special sponsorship provides the total prize money of more than £1,000 to winning students and their schools (see box) and contributes to the administration costs of the competition. In addition to the winners, each student entrant receives a certificate, a feature

which is highly valued by the schools and, therefore, worth the extra work involved for the MiSAC secretariat, generously provided by the Society for General Microbiology.

Officers of SfAM joined the Chairman and other members of MiSAC for the judging. The members of SfAM involved were General Secretary Anthony Hilton, Communications Officer Lucy Harper and Meetings Secretary Martin Adams, also a member of MiSAC. The judges looked particularly for attention to the guidance given to entrants on the creation of an information leaflet including originality, presentation and the effectiveness to communicate science to the general public.

Other important features were evidence of scientific merit, the use of entrants’ own words, e.g. not entire text taken directly from the web, and the appropriate use of illustrations to reinforce the message.

The standard of the entries was very high and students used a wide variety of media – from hand-produced drawings to high quality graphics giving a professional finish. Unfortunately, a few were excluded from consideration because they did not adhere to the rules of the competition.

But the numbers of excluded entries was lower than in previous years.

MiSAC and SfAM express their sincere thanks to everyone who took part, including those whose efforts were not rewarded with a prize on this occasion and the teachers who organised the preparation and submission of the entries.

We hope that the experience was rewarding and enjoyable, and has led to a greater interest in and understanding of microbiology.

The 2008 MiSAC competition will concern fungi and medicines, supported by special sponsorship from the British Mycological Society.

Key Stage 3 winners 1st: Hugo Fleming, Feltonfleet School 2nd: James Fryatt, St Aidans RC School 3rd: Katie Wensley, Sheffield High School Commended: Harry Hitchens, Prior Park College Commended: Emily Barnes, The Grange School

GCSE winners 1st: Daniel Tolley, St Anselms College 2nd: Christopher Lewis, Bedford School 3rd: Camille Raoult, The Abbey School Commended: Jawad Safdar, Bedford School Commended: Emily Kirkham, The Abbey School

John Grainger Chairman, MiSAC

September 2007

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members

Dr Wellyzar Sjamsuridzal of University of Indonesia Culture Collection (UICC) reports on the preservation of an endangered culture collection in Indonesia

Endangered Culture Collection Grant reports

Main photo: countryside and paddy fields near Jakarta. Inset left: Lunch at a traditional Indonesian restaurant in Depok. Inset right: Dr Sjamsuridzal examines a baby mango tree in a local garden centre.

Below: UICC members with Dr Peter Green (2nd from right)

further information The SfAM Endangered Culture Collection Fund is intended to help Society members to assist an endangered culture collection whose existence is under threat.

To apply for this award, please contact the Society Office or visit the website at: â&#x2013; www.sfam.org.uk/grants.php

48

T

he University of Indonesia Culture Collection (UICC) belongs to the Department of Biology, Faculty of Mathematics and Natural Sciences, University of Indonesia. It is unique in its holdings, which focuses on entirely indigenous Indonesian strains.

Since the establishment of UICC in 1980, it developed independently without any financial support from the government. Therefore, the collection could only maintain a limited number of cultures. UICC has a mission to preserve Indonesian microbial diversity.

It was registered as member of the World Federation of Culture Collections (WFCC) no.563 and holds a collection of bacteria, moulds and yeasts. It hosts around 2,000 isolates that mainly originated from environmental samples (e.g. soils, water, leaves, litter) and food products (e.g. salted fish, tuak, ragi, tape). At present, there are more than

September 2007

2,000 yeast isolates, 43 bacterial isolates, and 479 filamentous fungi in the collection. UICC has six part time scientific staff and one technician.

Problems The UICC houses the major yeast and Rhizopus collection in Indonesia from food-related and environmental habitats. Rhizopus is a cosmopolitan filamentous fungus found in soil, decaying fruit and vegetables, animal faeces, and old bread. While Rhizopus spp. are common contaminants, they are also occasional causes of serious (and often fatal) infections in humans.

Some species are plant pathogens.

Many of the Indonesian habitats for Rhizopus spp. have already disappeared due to deforestation, forestry plantations, floods and extensive fires in Indonesia.

Preservation methods are needed to ensure the high quality of cultures and their sustainability for use in basic and applied science, for example in the development of novel bioactive compounds, food science,

drying apparatus which was donated in 2001 by the Institute of Applied Microbiology (IAM) culture collection of the Institute of Molecular and Cellular Biosciences, University of Tokyo. We have started to use this apparatus for the L-drying method, but again no cold room is available to store the lyophilized cultures.

The grant from SfAM We are very grateful for the grant of ÂŁ2,500 from the Society for Applied Microbiology. This grant will be used for the establishment of a cold room which will guarantee the stability of the isolates and through this grant we can strengthen the role of our culture collections as national resource centres using high quality strains. This coupled with some spares for our freeze dryer is

a good start on the road to improving the sustainability of our valuable microbial resource.

Dr Peter Green visited the Department of Biology, Faculty of Mathematics and Natural Sciences, and during this time he was able to deliver a lecture on the Management of a Culture Collection and Preservation of Bacteria and Fungi. The seminar was attended by 146 people from universities, research institutes and industries in Jakarta, Depok, Bogor, Bandung, Purwakarta and Semarang. We are grateful to have had the opportunity to discuss our problems collectively and on a one to one basis with a Curator form a major national collection such as NCIMB. Dr Wellyzar Sjamsuridzal, UICC

Progress Report on the Chiangmai University Culture Collection which benefited from the SfAM Endangered Collection Grant in 2005

bioremediation, and biocontrol.

Unfortunately, the UICC cannot guarantee the medium to long term survival of these strains with the current methods of preservation.

Therefore, help is urgently required to improve our preservation technique, subculture. Currently this technique is the cause of many isolates dying and/or becoming contaminated. The increasing number of isolates that requires maintenance are stored in refrigerators which have reached their load capacity.

Up to now there is no regular budget for maintenance of these cultures. In addition to this, staff do not have time to subculture isolates due to other commitments at the University.

Therefore, a cold room is urgently required to guarantee the stability of the isolates, especially the fungal isolates, as a first step towards long term storage. We do have a basic freeze

Dr Green visited our laboratory in the Microbiology Division, Department of Biology, Faculty of Science, Chiang Mai University to follow up on current progress since his visit in 2005.

Since the SfAM grant, our Laboratory along with the culture collection, has moved to a new and more modern building on the same campus. With the new building we have much more space for culture collection and herbarium. Our cultures include endophytic fungi, actinomycetes and some endophytic bacteria which are used by both undergraduate and graduate students for their research.

Before the grant, our culture collection was in a dangerous condition since we did not have enough knowledge and funding to manage it effectively.

Now we have an improved database (including the materials stored in our herbarium of saprophytic fungi and wild mushrooms) and also improved storage and preservation facilities; although of course much more investment is still required.

Some important cultures are sent to the Biotechnology Culture Collection in Bangkok as backup, but the important strains are kept in our

September 2007

own Laboratory. As a result of the SfAM grant we have made progress in the above areas and have saved some cultures which would otherwise have been lost. Our collection of endophytic fungi, actinomycetes, (including Streptomyces and rare actinomycetes) are kept in glycerol broth and stored at -20C â&#x20AC;&#x201D; the standard method recommended by Dr Green. He also recommended keeping some of these strains freeze dried which we have yet to complete.

During Dr Greenâ&#x20AC;&#x2122;s visit we discussed methods of preparing and checking the quality of culture; both bacteria and fungi. Continued contact with Dr Green and other relevant WFCC experts helps our 19 PhD students and myself keep the culture collection in good condition.

As a result of our discussion we hope to hold a training course for other University personnel from the northern area of Thailand on how to manage small culture collections with limited resources. The key objective is to help us keep at least the most important strains in these collections alive to allow us to study them and benefit from their properties.

Professor Saisamorn Lumyong Chiang Mai University

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members

Students into Work Grant reports Photodynamic antimicrobial chemotherapy

information am I eligible can I apply?

Yes — if you are FULL member who can offer an undergraduate microbiology student the chance to obtain work experience. If you would like to read about the experiences of Students who have benefited from this Grant, you can do so in each issue of Microbiologist.

For Further information visit: www.sfam.org.uk/grants.php I am currently undertaking a pharmacy degree at the School of Pharmacy, Queen’s University Belfast. During the summer before my fourth, and final year, I availed myself of the SfAM Students into Work scheme. This has provided me with invaluable experience and knowledge of practical laboratory techniques, as well as the chance to explore the world of scientific research.

Having been extremely interested and intrigued by the topic of photodynamic antimicrobial chemotherapy, and in particular its application to the prevention of infection in cystic fibrosis patients, I decided that this was the area for me, so I approached Dr Ryan Donnelly whom I knew had a particularly keen interest in this area.

He was more than supportive to have me on board his group, and together we decided to investigate the susceptibility of clinical P. aeruginosa isolates growing planktonically and in biofilm to photodynamic inactivation using a combination of either toluidine blue O (TBO) or Meso-tetra (N-methyl-4-pyridyl) porphine tetra tosylate (TMP) and visible light.

Photodynamic antimicrobial chemotherapy (PACT) is a medical treatment by which a combination of a sensitising drug and visible light cause the selective destruction of microbial cells in the presence of oxygen. Photosensitising drugs, such as the modified porphyrins, phthalocyanines and phenothiazine dyes (TBO) have been shown to accumulate rapidly and selectively in bacterial cell membranes. Irradiation with a suitable dose of light of appropriate wavelength then elicits the photodynamic effect, killing the infecting microorganism and leaving host tissue unharmed. The group

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in this school have previously shown PACT to be a potentially novel treatment for infection by onychomycosis-causing pathogens.

Importantly, there have as yet been no reports on the development of microbial resistance to PACT. Recent evidence indicates that PACT may be useful in the treatment of infections caused by microorganisms resistant to conventional antimicrobial strategies. Moreover, this new technique has shown the ability to eradicate even microorganisms residing within multispecies biofilms. If PACT could be shown to be effective in the eradication of clinical isolates of P. aeruginosa, grown planktonically and in biofilm, then this may reduce the need for continuous use of antibiotics in cystic fibrosis (CF) patients and, therefore, help to prevent further antibiotic resistance developing.

Prior to determining the susceptibility of the microbial organism to the proposed photosensitisers (TBO and TMP) there was one important factor that we had to consider; would the drugs be able to effectively diffuse through the viscous mucus usually present in lungs of cystic fibrosis patients? Hence diffusion studies of the drugs were carried out.

Unfortunately, the results of these preliminary investigations were to no avail.

Firstly, no authentic cystic fibrosis sputum could be obtained, and the gels prepared to act as a model either themselves diffused across the modified Franz cell, or interfered with the UV analysis necessary to determine the amount of drug that diffused at any particular time. It is hoped that during my final year, I can build upon this knowledge and aim to overcome these problems so that an effective study into this important area can be assessed.

However, not one to be dishearted, I was eager to investigate whether the suggested photosensitisers could be used to kill P. aeruginosa (strain PAO1 NC1MB 10548). A study was undertaken to determine the optimum conditions for microbial death as a result of the TBO photosensitiser. It was found that : ■ There was no difference in kill with different incubation times at 370C ■ Interestingly the lowest concentration (0.05mg/ml) achieved the optimum kill These conditions were employed to investigate the effect of TBO and red light (635nm) on five other Pseudomonas aeruginosa strains (12A, 11D, 7-6087B, 11A and 6A). The highest percentage kill calculated was 99.9% for strain 12A and the lowest was 89.96% for strain 6A.

When investigating the susceptibility of the organism to the other photosensitiser, TMP, the method was slightly altered, due to the fact that the optimum concentration for kill was found to be 5mg/ml. Interestingly some strains of the organism where more sensitive than others to the drug’s action, with the percentage kill ranging from 99.77% for the 7-6087B strain, and 86.34% for the 6A strain. The difference in the precentage kill may be due to different strains being more susceptible to the uptake of the photosensitiser. For example, some strains may have efflux pumps, which they use to pump the drug out and away from the cell membrane, where these photosensitisers exert their effects.

Overall, as well as gaining invaluable experience in academic research , I believe that these results show promise for the future acceptability of PACT as a successful alternative for the management of infections in patients suffering from cystic fibrosis. We have highlighted that the Pseudomonas aeruginosa, so troublesome in many of these patients, is indeed susceptible to the action of the photosensitizing agents tested, and hopefully my research can be continued further to show that it may indeed be effective in a clinical setting.

Martin Garland Queens University, Belfast

The identification of Campylobacter spp. using an amplified fragment length polymorphism fingerprinting method When I arrived in Northern Ireland from the Czech Republic almost six years ago I had no idea that one day I would be taking part in (and enjoying!) a summer microbiology research project. In order to be accepted at university in the UK I had to complete all the required A levels and GCSEs as my original qualifications were not officially recognised here. I started again from scratch and after three years of concentrated studying, I was accepted into the Food and Nutrition BSc course at Queen’s University of Belfast which I will complete next year. Although I have always been very interested in nutrition, I really enjoyed the food microbiology course. I have realised how closely food microbiology is linked to human diet and health. I did not hesitate when an opportunity arose to work on a microbiology project in the Food Microbiology Branch of the Agri-Food and Biosciences Institute (AFBI) in Belfast.

Human isolates of Campylobacter spp. from Sweden were investigated for the study.

Amplified fragment length polymorphism (AFLP) fingerprinting was used to detect polymorphisms in the genome. The method involved digestion of DNA using two restriction enzymes (Bgl II and Csp 61), ligation of the adapters (frequent and rare) to the sticky ends produced by restriction enzymes, and direct selective Polymerase Chain Reaction (PCR) amplification of selected fragments (Kokotovic and On, 1999).

The samples were run on an ABI 3100 sequencer. The AFLP profiles were analysed using Bionumerics 4.50 software. Samples were classified into clusters and expressed using dendrograms. The clusters are calculated based on the similarity of each of the samples DNA fragment sizes. If the similarity between samples is over 90% they are considered to be identical, based on the control samples.

All 76 samples were identified as Campylobacter jejuni. From the dendrogram it was obvious that two significant clusters were obtained: cluster 1 containing 42 isolates and cluster 2 with 24 isolates. The remaining isolates (n = 10) showed less than 80% similarity to the two clusters. Isolates in cluster 1 and 2 came as close as 58.4% and 67.4% respectively.

Although all the isolates were Campylobacter jejuni, there was genetic diversity within the groups. In cluster 1 there were three main groups. Group 1 (n = 7) clustered at 83.1% similarity, group 2 (n = 12) at 82.3% and group 3 (n = 6) at 86.4%. Group 2 was very interesting as eight isolates were identical (more than 90% similarity), suggesting that the cases had a common source. Identical isolates also occurred in the groups 3, 4 & 5. In total 44.7% of the isolates showed similarity over 90% with at least one other isolate.

In the original study Höök et al (2004) used information such as date and place of

isolation, sex and age of patients and the pulsed-field gel electrophoresis (PFGE) type, in order to find out if there was a pattern in the distribution. It did not seem that there were any obvious patterns in distribution.

Forty samples were isolated from women and 36 from men. Most of the patients (n = 48) were ill during the summer months and the most frequent location was Västerås, the main city in the area. People of all ages were ill. The analysis of the AFLP profiles will be forwarded to Dr. Helena Höök from Swedish University of Agricultural Sciences who will compare them with analysis of PFGE types she had determined as part of her doctoral research project.

This work experience has broadened my horizons and provided me with a great range of skills. I have also learned how to manage my time, plan experiments, use all the available resources such as journals and textbooks effectively, and also how to work as part of a team. I would like to thank Dr. Robert Madden and Mrs. Moran for sharing their knowledge and expertise from their respective areas. Also Mrs Carmel Kelly for keeping me on the right track and making my summer work really enjoyable. After this experience, I would certainly consider doing further research in the future, such as a PhD, in the area of food microbiology or nutrition. Finally, I would like to thank the Society for Applied Microbiology for enabling me to carry out this project. It has been a very valuable experience!

September 2007

References ■ Kokotovic, B. and On, S.L.W. (1999) High-resolution genomic fingerprinting of Campylobacter jejuni and Campylobacter coli by analysis of amplified fragment length polymorphism. FEMS Microbiology Lett. 173, 77-84 ■ Höök, H., Ekegren, M., Ericsson, H., Vågsholm, I. and Danielsson-Tham, M-L. (2004) Genetic and epidemiological relationship among Campylobacter Isolates from Humans. Scand. J. Infect. Dis. 36, 435-442

Martina Strawbridge Klapkova

Not a student member?

For full information on the many benefits available to SfAM student members, turn to page 6

51

members

President’s Fund reports information am I eligible can I apply?

If you often find difficulty in funding attendance at meetings.

It is not only our student members who require our help.

Senior microbiologists often find difficulty in funding attendance at meetings. If you are in this position you are eligible for this fund.

For Further information visit: www.sfam.org.uk/grants.php

52

Novel Decontamination Methods Decontamination of food, the environment and food contact materials are of major concern to the food manufacturer and such operations must be risk based so that the resources are targeted appropriately. Recent years have seen an expansion of the technologies that can be used for decontamination. This article will outline some of these novel techniques.

Ozone has long been used for the treatment of water and many drinking water facilities in France employ ozone (Gurley 1985). Ozone can retain its high oxidation potential (+0.27 volts) when dissolved in water, allowing oxidization of microbial cells. Ozonated water is effective in the cleaning of beer lines (Fielding et al., 2007) and stainless steel surfaces. In Europe, ozone and ozonated water may have potential in replacing traditionally used cleaning agents as they leave no residues and, as they are produced in situ, are exempt from the Biocidal Products Directive (98/8/EC). Ozone has been used to reduce the general microbial levels in a cheese making factory when applied overnight after normal cleaning operations. It is also used to decontaminate the stand-alone water coolers. Ozonated water has lower health and safety implications than the use of ozone gas and can be produced a short time in advance of use.

Open Air Factor (OAF) was first described in 1968 and, despite extensive research, very little information was published due to the nature of the research (National Defense) (Druett & May 1968, 1969). OAF is the highly reactive chemical species produced when ozone reacts with any compound containing a carbon–carbon double bond. It is not a single molecule but a collection of chemical species that can vary depending on the unsaturated organic molecule with which ozone reacts. This early research indicated that bacterial survival in aerosolized particles was much greater in a closed vessel than in the open air (the external environment). The potential of artificially generated OAF as an air phase and surface disinfectant has only recently been considered and has proved to be effective for reducing the levels of Micrococcus luteus in air (Bailey et al., 2007).

Electrolysed Acid Water (EAW) is an electrochemical solution produced by electrolysis of water containing approximately 20% sodium chloride. This produces hypochlorous acid and hydroxyl radicals that have a rapid bactericidal effect. Electrolysed Acid Water has a strong oxidizing ability, a redox potential of more than 1000mV, 30 ppm dissolved chlorine and a pH lower than 2.7. There are two types of EAW — strong (ESAW) and weak (EWAW). Weak is highly corrosive and not suitable for use in the food industry. Strong is suitable and has antimicrobial properties in narrow bore tubing systems and against a wide range of microorganisms. The EAW is produced at a site close to the point of use and requires only potable water and sodium chloride as raw materials. It degrades to form saline and trace chlorine gas and is of minimal risk to humans and the environment. Chlorine dioxide is becoming an increasingly popular sanitizer as, unlike sodium hypochlorite, no trihalomethanes or chlorophenols are produced when it is used. It is active over a wide pH range, with best antimicrobial activity at a slightly alkaline pH. It is not inhibited by food residues (unlike ozone or ozonated water) and is very effective against spore-forming organisms. It is slightly more potent than chlorine as a disinfectant so can be used at lower concentrations than chlorine and has fewer problems regarding tainting of food. It has found application in water treatment, the brewing, poultry and produce industries as well as in food contact paper manufacture.

Cleaning of food raw materials is often necessary prior to further preparation or processing.

While some of the methods are similar to those used for surfaces and equipment, care must be taken to avoid damage to the food. If the presence of residual chemical on the food is problematic, ultrasonic cleaning may be an option whereby sound waves (≥ 20 000 vibrations per second, leading to rapid formation and then collapse of bubbles in the fluid [cavitation and decavitation]) cause disruption to cellular structure and function, leading to cell lysis.

Ultraviolet light has been explored within a wide number of areas of the food industry including poultry processing, chill rooms for meat storage and chocolate manufacture. It is proving increasingly popular in the water industry, especially the vending industry. The fact that no toxic by-products remain gives UV light an advantage over chlorine-based methods. The use of UV in disinfecting water is facilitated by the relative lack of other absorbing substances, as these can reduce its effect by absorbing some of the radiation. The effect of UV varies between different microorganisms with spores, cysts and viruses having a higher resistance. One disadvantage of this technology is its low penetration power, meaning that it can only be used as a surface treatment. Crevices in the food or a surface that is not directly in line with the UV source will lead to protection of microorganisms. High intensity pulsed light involves the use of intense, short duration pulses (a fraction of a second) of ‘white light’ (the visible range of the spectrum, between UV and infra-red). This technique has been utilised in the decontamination of packaging surfaces. Cleaning and sanitation are the major control measure associated with surface hygiene, whether these surfaces are environmental, food and hand contact surfaces or the food itself. There are a number of factors, however, that can lead to failure of the sanitation programme and allow soil to accumulate and microorganisms to survive and multiply. It is essential that the efficacy of decontamination procedures are continually monitored, assuring the producer that there is a minimal risk of product contamination and that the processing premises are being operated safely and hygienically.

September 2007

References ■ Bailey R., Fielding L., Young A. and Griffith C. (2007) The effect of ozone and ‘Open Air Factor’ against aerosolised Micrococcus luteus. Submitted to the Journal of Food Protection.

■ Druett H. and May K. (1968) Unstable germicidal pollutant in rural air. Nature (London) 220, 395-396 ■ Druett H. and May K. (1969) The Open Air Factor. New Scientist 41, 579-581 ■ Fielding L.M., Hall A. and Peters A.C. (2007) An evaluation of ozonated water as an alternative to chemical cleaning and sanitization of beer lines. Accepted for publication, Journal of Food Service.

■ Gurley, B. (1985) Ozone: Pharmaceutical sterilant of the future? Journal of parenteral science and technology, 39 (6), 256-261

Louise Fielding, University of Wales Institute, Cardiff

Metal ion homeostasis and Salmonella pathogenicity Salmonella spp. are Gram-negative enteropathogenic bacteria responsible for more than a billion cases of human disease each year, with clinical manifestations ranging from gastroenteritis, enteric fever and systemic typhoid fever. S. enterica serovar Typhimurium (S. Typhimurium) is a major cause of gastroenteritis in humans, rarely causing systemic disease. However, in mice, this organism causes similar symptoms and disease progression to human typhoid fever and hence has been intensely studied as a model for this disease. In this model, following ingestion the salmonellae colonise the ileum and caecum and are translocated across the intestinal epithelium via invasion of the M cells, or through uptake by dendritic cells, and are spread to target organs such as the liver and spleen (recently reviewed by Coburn et al., 2006). The ability of S. Typhimurium to survive within macrophages is critical during the systemic stages of disease.

Within macrophage phagosomes, S. Typhimurium must sense and respond rapidly to a wide range of fluctuating environmental conditions, including changes in the availability of essential nutrients and exposure to reactive oxygen species. Transition metal ions (such as iron, copper and zinc) are required by bacteria as essential cofactors for a large number of proteins, including those involved in respiration, electron transport, macromolecular synthesis and detoxification of reactive oxygen species. However, while these metal ions are essential they are also toxic in excess and intracellular concentrations of each particular metal must be precisely controlled.

Bacteria, including Salmonella, therefore require efficient metal homeostatic mechanisms to avoid metal mediated stress. Indeed, genes encoding proteins involved in the sensing and transport of various metal ions are known to contribute to S. Typhimurium virulence and are among the major genes expressed during infection of macrophages (Heithoff et al., 1997, Eriksson et al., 2002).

Susceptibility of mice to infection by S. Typhimurium has been shown to depend on the presence or absence of a functional allele at the ity/lsh/bcg locus (Vidal et al., 1993). Resistance to other intracellular pathogens such as Mycobacteria spp. and Leishmania spp. is also dependent on this locus and lack of a functional gene may alter the ability of macrophages to destroy these intracellular pathogens (Searle et al., 1998). The gene product of this locus, Nramp1 (natural resistance associated macrophage protein 1) (also referred to as Slc11a1; solute carrier family 11, member a1), is an integral membrane protein expressed almost exclusively in macrophages and in particular in lysosomal compartments (Grunheid et al., 1997, Searle et al., 1998). The mechanism by which Nramp1 controls bacterial replication in macrophages is not fully understood although in vitro studies have revealed it acts as a proton/divalent cation antiporter (Goswami et al., 2001) and is capable of transporting (at least) Fe2+, Mn2+ and Zn2+ (reviewed in Courville et al., 2006). Suggested roles include exporting ferrous ions from the phagolysosomal compartment in order to ‘starve’ the pathogen of an essential nutrient and, in contrast, supplying ferrous ions to this compartment to drive the Fenton reaction and generate highly toxic hydroxyl radicals. It is possible that phagosomal metal levels are dynamic, switching more than once at different stages during the maturation of the phagosome, and pathogens which inhibit this maturation may evade some of these challenges.

The metal stresses encountered by S. Typhimurium during infection and within macrophage phagosomes are not known.

Historically, much of the Salmonella in vivo models of infection have used the BALB/c mouse which carries a mutation at the ity/lsh/bcg locus and hence are Nramp1-/-. Similarly, in vitro models of infection using cultured macrophage cell lines such as J774A.1 and RAW264.7 are also naturally Nramp1-/-. It is anticipated that the presence or absence of Nramp1 will influence the availability of certain metal ions within the phagosome, although the metal substrates in vivo remain to be resolved.

Previous studies in our laboratory have shown that metalresponsive promoters can report cytosolic metal levels, and hence metal stresses, as sensed by a bacterium (Cavet et al., 2002). Indeed, we can correlate these changes in promoter expression with environmental metal levels. We have precisely calibrated the metal specificities of a number of metal-responsive promoters and are now using this panel of promoters to directly monitor changes in the levels of iron and other metals encountered by S. Typhimurium during phagosome development (in Nramp1+/+ and Nramp1-/macrophages). Our aims being to resolve (i) whether a bacterium is exposed to elevated or depleted metal levels in phagolysosomes, (ii) which are the predominant metals that fluctuate in concentration (as sensed by S. Typhimurium) and when the fluctuations occur, and (iii) the principle substrates of Nramp1 and direction of transport. By understanding the metal stresses that S. Typhimurium is exposed to within macrophage phagosomes, we hope to gain an insight into the mechanisms underlying Salmonella survival in the host environment to inform future control strategies and treatments.

References ■ Cavet, J.S., Meng, W., Pennella, M.A., Appelhoff, R.J., Giedroc, D.P. and Robinson, N.J. (2002) A nickelcobalt-sensing ArsR-SmtB family repressor. Contributions of cytosol and effector binding sites to metal selectivity. Jnl Bio Chem. 277, 38441-8 ■ Coburn, B., Guntram, G.A. and Finlay, B.B. (2006) Salmonella, the host and disease: a brief review. Imm and Cell Biology. 5, 1-7 ■ Courville, P., Chaloupka, R. and Cellier, M.F.M. (2006) Recent progress in structure-function analyses of Nramp proton-dependent metal-ion transporters. Biochemistry and Cell Biology, 84, 960-978 ■ Eriksson, S., Lucchini, S., Thompson, A., Rhen, M. and Hinton, J.C. (2003) Unravelling the biology of macrophage infection by gene expression profiling of intracellular Salmonella enterica. Molecular Microbiology. 47,103-18 ■ Goswami, T., Bhattacharjee, A., Babal, P., Searle, S., Moore, E., Li, M., and Blackwell, J.M. (2001) Naturalresistance-associated macrophage protein 1 is an H+/bivalent cation antiporter. Biochemical Journal. 15, 5119 ■ Grunheid, S., Pinner, E., Desjardins, M. and Gros, P. (1997) Natural resistance to infection with intracellular pathogens: the Nramp1 protein is recruited to the membrane of the phagosome. Journal of Experimental Medicine. 185, 717-730 ■ Heithoff, D.M., Conner, C.P., Hanna, P.C., Julio, S.M., Hentschel, U. and Mahan, M.J. (1997) Bacterial infection as assessed by in vivo gene expression. Proceedings of the National Academy of Science U S A. 94, 934 -939 ■ Searle, S., Bright, N.A., Roach, T.I., Atkinson, P.G., Barton, C.H., Meloen, R.H. and Blackwell, J.M. (1998) Localisation of Nramp1 in macrophages: modulation with activation and infection. Journal of Cell Science.111, 2855-2866 ■ Vidal, S.M., Malo, D., Vogan, K., Skamene, E. and Gros, P. (1993) Natural resistance to infection with intracellular parasites: isolation of a candidate gene for BCG. Cell 73, 469-485

Clare Taylor, University of Manchester

T2 Scientific presents ‘Methods for the Examination of Water & Associated Materials’ by kind permission of the Standing Committee of Analysts ■

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TCS Biosciences Ltd collaborates with HPACC We are delighted to announce our collaboration with the Health Protection Agency Culture Collections. TCS Biosciences Ltd has agreed a Manufacturing License with HPACC, with the result that Selectrol® will be manufactured exclusively from NCTC and NCPF strains.

Gareth Williams, Sales and Marketing Director of TCS Biosciences Ltd commented: “We are very excited about our collaboration with HPACC. This Manufacturing License underlines our commitment to providing a multipurpose Selectrol® product that meets the requirements of our accredited customers in clinical, food and industrial laboratories.”

Selectrol® is a convenient source of viable micro-organisms which can be used for a variety of quality control and testing applications that are guaranteed to be a first generation derivative of the original strain.

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September 2007

We are pleased to notify our customers of the addition of Salmonella nottingham NCTC7832 to the Selectrol® range.

Salmonella nottingham is recommended by the HPA as the positive control in the detection of Salmonella sp. Salmonella nottingham NCTC7832 is now recommended as a result of increased prevalence of the previously recommended Salmonella poona in food samples. further information visit: www.tcsbiosciences.co.uk

Guidance on microbiological criteria for food products from CCFRA A new guideline document — Establishment and use of microbiological criteria (standards, specifications and guidelines) for foods (Guideline 52) — is now available from CCFRA. The value of meaningful microbiological criteria in the safe production of food is a major interest of the food and drink industry. Microbiological testing of food cannot control the safety of the food, but can be used for a variety of purposes, including the support and verification of HACCP. In order to be able to set a microbiological criterion for a foodstuff, it is useful to have a HACCP system covering the production of that foodstuff.

Written by professional food microbiologists with experience in the food manufacturing, retailing and testing sectors, this guideline sets out the current thinking with respect to microbiological criteria (microbiological specifications, microbiological guidelines and microbiological The latest news, views and microbiological standards). It discusses how microbiological developments from our corporate members specifications and guidelines can be developed and suggests best use of the resource and resulting data when testing food for the presence of microorganisms. It aims to provide guidance to all those involved in producing and using microbiological criteria in the food and catering industries.

corporate news

Sales Director at DWS, Steve Robertson, commented: “The miniMACS was initially developed to fulfil a need for anaerobic and microaerophilic workstations where either space is limited or sample throughput makes it harder to justify the purchase of a larger capacity unit.”

The miniMACS workstation has an impressive 33% more capacity than any other unit of comparable size. When used solely as an incubator, as many as 400 x 90mm Petri dishes can be accommodated.

Don Whitley created the first anaerobic workstations in the UK and invented the variable atmosphere (VA) cabinets in the early 1980s. Now the miniMACS has become the fastest selling product in the entire MACS workstation range and this particular order marks a milestone in the company’s history. It is the largest single order for MACS workstations DWS has received to date. further information visit: www.dwscientific.co.uk Tel: +44(0)1274 595728

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MiniMACS to China It’s not quite ‘coals to Newcastle’ but supplying home engineered and manufactured goods from the UK to China usually happens the other way around these days. Don Whitley Scientific Limited (DWS), however, has recently dispatched 21 miniMACS workstations to a customer in China.

Lab M’s new Baird-Parker Medium (ISO) has been developed to meet specific ISO requirements in the food industry. This improved version of Baird-Parker Medium, for the isolation of coagulase-positive staphylococci including Staphylococcus aureus from food and animal feedstuffs, is formulated according to ISO 68881:1999 and is in compliance with ISO 68882:1999 and 6888-3: 2003. It is tested in accordance with ISO/TS 11133-2:2003(E).

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September 2007

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information Are you a corporate member of the Society?

If so, this section of Microbiologist is for you. Here you can publish short press releases, acquisition notices, news of new staff appointments, technical developments and much more.

Each corporate member of the society may publish up to 200 words on a topic related to their field of activity in each issue of Microbiologist. For further information please contact Lucy Harper by email at: lucy@sfam.org.uk Both corporate members and ordinary members of the Society will find a wealth of useful information and resources in this section.

58

materials, Lab M’s Egg Yolk Tellurite has been improved to complement this Baird-Parker formulation. As with Lab M’s traditional BairdParker medium, the new formulation can be used with either Egg Yolk Tellurite or Rabbit Plasma Fibrinogen (RPF). The use of RPF produces enhanced reactions compared with previous BairdParker formulations.

When the medium is used with Egg Yolk Tellurite, presumptive S. aureus appear as black colonies demonstrating lecithinase activity as an opaque zone around the colony, encircled by a zone of clearing that is the result of lipase activity.

Coagulase (RPF media) or latex agglutination tests are used for confirmation.

Where Rabbit Plasma Fibrinogen is used, typical S. aureus appear as black colonies surrounded by a zone of precipitation, indicating coagulase activity Further confirmatory tests are not necessary.

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Anaerobic Developments With an increased incidence of serious Anaerobic infections, in particular with Clostridium Difficile, the need for innovation in the field of development of Anaerobic Workstations is of paramount importance.

This need is clearly recognized at Ruskinn Technology and Pro-Lab Diagnostics (Exclusive agents in the UK), where a dedicated team of professionals have developed the most advanced, ergonomically designed Anaerobic Workstations available. A range of workstations is available from the space saving, economical Bug Box for handling up to 200 plates, to the Concept 400 range, through to the Concept 1000 for up to 1000 plates with the facility for two operators to work side by side.

Standard available features for the workstations include single plate entry systems, dual gas operation, low gas consumption, rapid interlock chambers, low gas alarms, energy saving lighting, humidity control, excellent capacity to footprint ratio, uniquely designed port entry systems, foot operated controls, colour coded petri dish holders and much more.

In addition to these standard features, workstations can be assembled to customer specific specifications and even have equipment installed such as mixers, platers, microscopes and September 2007

Oxoid launches robust new automatic air sampler The new robust, high specification Oxoid Air Sampler is designed to monitor microbiological air quality in controlled pharmaceutical, clinical and food processing environments, using proven impaction technology.

Compact and easily portable, the sampler is easy to use. The menu driven screen leads you through preset or user-defined functions, including flexible sampling volumes, delayed start, multiple run and variable interval options. It can be operated manually or via infra-red remote control — ideal for monitoring highly sensitive, closed environments.

In validation studies the sampler demonstrated a physical efficiency of 100% for particles 0.8 to 19.0 microns in size (BS EN ISO 14698-1:2003 Annex B). Air is sampled at a rate of 100 litres per minute. Sample details (eg date, time, location, sampled volume and operator ID) are stored within the unit; for reporting/trend analysis results can be downloaded to a computer using the optional Oxoid Air Sampler Software.

The unit is supplied fully calibrated and the sample head can be changed, or removed for cleaning, quickly and easily without the need for recalibration. further information visit: www.oxoid.com Tel: +44 (0) 1256 841144 Email val.kane@thermofisher.com

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Solutions for Industrial & Clinical Microbiology

INNOVATION IN FOOD MICROBIOLOGY

Inhibigen™ Technology: A NEW DIMENSION IN SELECTIVE MICROBIOLOGY Oxoid has developed a unique, new class of selective agents known as Inhibigens™. When added to a culture medium, these molecules provide highly specific selectivity and allow improved recovery of often stressed target organisms.

Inhibigen™ technology, which is currently subject to a patent application, involves the use of an inhibitor molecule linked to a specific substrate. In this bound state, the inhibitor is non-toxic - however if taken up by a cell and cleaved from the substrate, the inhibigen molecule will prevent the organism from replicating. Only organisms with the required uptake mechanism and specific enzyme to cleave the inhibitor substrate complex will be affected. This allows very specific inhibition of competing, non-target organisms.

Unlike conventional selective agents, such as antibiotics, Inhibigens™ can be engineered to have no inhibitory effect on the target organism - even when the cells are stressed. This means that recovery of specific organisms is improved in two ways - by reducing the growth of competing flora and by minimising exposure to potentially inhibitory components.

Oxoid Salmonella Chromogenic Medium Mark II (OSCM II) is the first ever selective culture medium to incorporate Inhibigen™ technology, combining it with familiar chromogenic technology to provide excellent isolation and identification of Salmonella colonies.

The Inhibigen™ used in OSCM II specifically inhibits the growth of E. coli, a common competing organism in Salmonella investigations, whilst novobiocin and cefsulodin at carefully selected levels, inhibit the growth of other competing flora, such as

Proteus and Pseudomonas. Two chromogens are also added to the medium, allowing the differentiation of Salmonella colonies (bright purple) from other

The R&D team responsible for Oxoid’s new Inhibigen™ technology

organisms, such as Klebsiella and Enterobacter (blue).

The combination of these principles makes it easier to identify Salmonella colonies and reduces the number of false positive results requiring follow-up investigations. With OSCM II you can experience a new level of efficiency in your laboratory. For more information please speak to your local Oxoid representative or contact Val Kane, Oxoid Ltd, on 01256 841144, email: val.kane@oxoid.com Inhibigen™ technology - a new dimension in selective microbiology.

Oxoid Limited, Wade Road, Basingstoke, Hampshire, RG24 8PW, UK Tel: +44 (0) 1256 841144 Fax: +44 (0) 1256 329728 Email: val.kane@oxoid.com www.oxoid.com