The Australian Research Institute Annual Report 2001 T

Council Members Registered office

Mr D.J. McWilliam, BSc, MSc Waite Road, Urrbrae, SA 5064 Chairman—Elected a Postal Address: member under Clause 6(e) PO Box 197, Glen Osmond, SA 5064 of the Articles of Association t (08) 83 03 66 00 f (08) 83 03 66 01 Mr R.E. Day, BAgSc, BAppSc Elected a member under Clause 6(e) of the Articles The Company of Association The Research Institute was incorporated under Mr R.L. Gibson, the South Australian Companies Act on 27 April 1955. It is BAppSc, BAppSc a company limited by guarantee, it does not have a share Elected a member under capital and it has been permitted, under licence, to omit the Clause 6(e) of the Articles word ‘limited’ from its registered name. of Association The Memorandum of Association of The Australian Wine Mr P.F. Hayes, Research Institute sets out in broad terms the aims of the BSc, BAppSc, MSc, DipEd Institute and the Report of the Committee of Review for the Elected a member under Institute published in March 1977 identified the following Clause 6(e) of the Articles specific aims: of Association 1. To carry out applied research in the field of . 2. To service the extension needs of the winemakers of Australia. Professor P.B. Høj, 3 To be involved in the teaching of oenology at both MSc, PhD undergraduate and postgraduate levels. Ex officio under Clause 6(d) 4. To assume responsibility for the co-ordination of oenological of the Articles of Association activities, and the collection, collation and dissemination as Director of the Institute of information on oenological and viticultural research to the benefit of the Australian wine industry. Mr T.W.B. James, AssDip(WineProd) The Institute’s laboratories and offices are located on the Elected a member under Waite Campus of The University of Adelaide at Urrbrae in Clause 6(e) of the Articles the Adelaide foothills, on land leased from the University. of Association The original lease is for a term of 99 years, with a right of renewal clause for a further 99 years. The Institute Mr G.R. Linton, formally affiliated with The University of Adelaide in 1990. BAppSc(AppChem), The first buildings were erected and opened in 1957 and GradDip(SysAnal) alterations and extensions were completed in 1976. The Elected a member under buildings have been extensively modified and refurbished Clause 6(e) of the Articles since that time with major extensions being undertaken in of Association 1994 and 1999.

Professor G.R. Scollary, The Institute is adjacent to the Faculty of Agricultural and MSc, PhD, BEd, BAppSc Natural Resource Sciences of The University of Adelaide, (Wine Science), FRACI three Divisions of the CSIRO and the South Australian Charles Sturt University Research and Development Institute. Representative under Clause 6(c) of the Articles of Association

Professor M. Sedgley, PhD The University of Adelaide Representative under Clause 6(b) of the Articles of Association

Dr R.R. Walker, BAgSc(Hons) PhD CSIRO Representative under Clause 6(a) of the Articles of Association

2 TTable of contents

2 Chairman’s report

3 Council notes

4 Director’s report

6 Staff

7 Highlights of the year

8 Staff activities

11 Research Teams’ reports

52 Appendixes 1–4

55 Abridged financial report

46th Annual Report Presented to the Australian Wine Industry 30th June 2001 Chairman’s Report

Doug McWilliam, Chairman of Council

It is my pleasure to again present my Chairman's rapidly changing and advancing technology appointed Professor of . Thank you Report for the past year's activities of The of closure design. to all the staff who carry the Institute's messages Australian Wine Research Institute. This past to industry so well via the forums of roadshows, year has been full of activity and outstanding One of our aims has been to strengthen the seminars, workshops, lectures and our progress with results now issuing from such chemistry side of our activities and we have publication, Technical Review. Congratulations projects as the closure trial, the potential been fortunate to have Dr Markus Herderich to our Communication and Publicity Manager enhancement of flavour during join our Institute staff. Markus undertook post and the Library staff for the implementation ageing and the ability of NIR to rapidly doctoral studies at our Institute in 1994-95 of the on-line searchable database of the measure berry colour and other quality and has strengths in flavour and phenolic John Fornachon Memorial Library. parameters. This last project has generated chemistry. Other major staff changes are the enormous industry interest and has not only retirement, after 14 years service, of our Finally, I offer my thanks to the Director, seen generous industry support from some of Secretary, Janet Currie. She not only kept a Peter Høj, and all of his staff for a successful our major companies but has now progressed tight reign on the financial affairs of our year. It has indeed been a pleasure for me to far enough to be trialled with confidence at Institute but was a counsel and confidant to have been associated with the Institute and the weighbridge. The NIR project all of the staff. We now have a most suitable its employees in my capacity as Council started as a collaborative project between the replacement and welcome Hans Muhlack to member and Chair. I urge all members of Institute and BRL Hardy but has now been fill the vacancy created by Janet’s retirement. our industry to not only continue, but incorporated in the new Cooperative Research increase, their interest and support for this Centre for Viticulture's programs and along The new extensions to the Institute are fully wonderful asset we all share. with the direct industry investment has led occupied and Analytical Services has continued to a strong industry desire to progress the to expand their range of services offered to development of affordable instrumentation the industry during the last year. The Institute that can be readily taken up by industry. The urgently needs to address the renovation and commercialisation of the IP associated with extensions of the old chemistry lab. With a this technique needs to be rapidly implemented stronger emphasis on flavour and phenolic and it is hoped to obtain prompt and satisfactory chemistry, these facilities are inadequate for agreement with all vested interests. the future needs of both equipment and staff. Doug McWilliam The closure trial, now more than 18 months It is pleasing to see that the Grape and Wine Chairman of Council old, has revealed significant differences in the Research and Development Corporation is performance of different closures as outlined proposing to move to a ‘rolling’ funding cycle in the publication which was due to appear which will remove some of the uncertainty in the July issue of the Australian Journal of of the current annual funding. It has been of Grape and Wine Research. What is most mutual benefit to have their representative important to recognise is that the testing attend our Council meetings and I thank them protocol and the individual results have to be for their advice, interest and involvement. evaluated from each winemaker's perspective of what performance factors they deem I also thank all members of Council for their important for their individual situation. The diligence and efforts with a special thank you Institute trial was a massive undertaking to Professor Margaret Sedgley from The involving considerable cost and resources University of Adelaide who retires in favour and only provides a ‘snapshot in time’ of the of Professor Steve Tyerman, the newly

2 Council notes

Chairman At the Council Meeting held on 30 October 2000, Mr D.J. McWilliam was re-elected Chairman of Council.

Members of the Executive Committee Mr D.J. McWilliam Mr R.L. Gibson Mr P.F. Hayes Professor P.B. Høj Mr T.W.B. James Professor M. Sedgley

Deputy Members of Council Mr N.P. Blieschke Mr L.P. Deans Dr P.R. Dry Mr J. Northey Dr N.S. Scott Dr A.M. Smith Mr P.J. Wall

Meetings Ordinary General Meeting The 46th Ordinary (Annual) General Meeting was held on 30 October 2000.

Council The Council of the Institute met on the following dates: 25 July 2000, 30 October 2000, 6 March 2001, 1 May 2001. Executive members of Council met on 5 December 2000.

Funding The Council of the Institute acknowledges the continuing financial support of the Grape and Wine Research and Development Corporation.

Appreciation The Institute acknowledges the assistance and cooperation of the following organizations throughout the year:

Adelaide University Australian Wine and Brandy Corporation Australian Wine Foundation Charles Sturt University Commonwealth Scientific and Industrial Research Organization (CSIRO) Cooperative Research Centre for Viticulture Left to right: Department of Agriculture, Forestry Peter Hayes, Robin Day, Geoff Linton, and Fisheries Australia Doug McWilliam, Margaret Sedgley, Richard Gibson, Steve Tyerman, South Australian Wine and Brandy Tim James, Peter Høj. Industry Association State Departments of Agriculture Winegrape Growers’ Council of Australia, Inc. Winemakers’ Federation of Australia Incorporated

3 Director’s Report

The continued growth of the Australian wine our trained sensory panel. This event precipitated hardware and different yeasts. The are industry and the increase in research levies, with unprecedented national and international now subjected to detailed sensory analyses effect from the 1999 , has allowed media interest. Throughout this process, and outcomes will be communicated to for an expansion in the total research portfolio relevant Institute staff conducted themselves industry upon its completion and presented managed by the Grape and Wine Research with a high degree of professionalism in an to industry in workshops conducted at the and Development Corporation (GWRDC) and , area which, due to its commercial significance, 11th Australian Wine Industry Technical in turn, for an expansion of funding to a had to be navigated with a great deal of Conference in October 2001. wide range of different research organisations. discipline and thought. Another highlight The funds allocated directly and indirectly to relates to the much awaited installation of The 11th AWITC will be the largest and the Institute have also increased in dollar- six computer-controlled Potter fermentors probably most diverse ever conducted. The terms and now constitute about 35% of the and six rotofermentors in the recently built Conference Planning Committee is Chaired total matched levy funds administered by the Hickinbotham-Roseworthy Wine Science by the Institute Director, the Program Planning GWRDC and an estimated 16-18% of public Laboratory at the Waite Campus of Adelaide Committee by Southcorp’s National expenditure on grape and wine R&D in University. This facility and its eventual Viticulturist (Peter Hayes) while the detailed Australia. We are appreciative of this funding completion is a prime example of how management is undertaken in a highly as it has widened the scope of our activities relentless belief in a cause and distinct professional manner by the Institute’s Rae Blair and further reduced our already very low per contributions by a range of determined and her Secretariat Staff. This triennial event capita running costs to service the needs of individuals can lead to long term gains where has grown to become a major Institute activity industry better. These activities will, in turn, others might have given up due to lack of in the area of knowledge dissemination and help underpin sustained growth of wine support. The Institute is most appreciative of adoption. It involves detailed planning, industry activities with consequential flow-on the University of Adelaide’s decision to build coordination and management of nearly benefits to local communities. this facility and indeed helped start the ball 1700 delegates, the staging of 72 rolling by pledging a $250,000 contribution workshops (coordinated by our Industry Tangible outcomes, about which much is over five years starting in July 1997; a Services Personnel) and arranging some written in the remainder of this Annual commitment we now have met fully due to 10,000 m2 of trade exhibition comprising Report, include the conduct, data collection, the strong performance of our own Analytical 178 individual exhibitors. analyses, writing up and acceptance for Service and its fourteen staff. In return for publication in the Australian Journal of Grape this significant contribution, we have been The selected examples of Institute activities and Wine Research of the Institute’s seminal granted access to the Facility for 25 years. cited above illustrate the continued closure trial, ably led by our Manager of The first such substantial access occurred commitment of our staff to the wine Industry Services (Peter Godden) with strong through a collaborative effort in our tannin industry which, in many cases, extends support from our sensory science specialist project where eight tonnes of fruit were beyond what can reasonably be expected (Leigh Francis) and the dedicated members of fermented in triplicate with different and sustained in the longer term without

Peter Høj

4 increased funding to employ more staff and Australian Government (including a R&D needs. A special thanks must go to Janet upgrade infrastructure significantly. The $700,000 cash contribution) and from Currie, our long-serving Company Secretary Institute is working actively to help itself in Council members who endorsed the Director who, over a period of 14 years, steered the this regard. One such example relates to the to commit to a further commercial loan in financial activities of the Institute without a large extension of Institute facilities we order to be in the race. A special mention flinch and allowed us to expand the Institute undertook in 1998. That $1.1 million extension should go to Mr Robin Day who, on several in a sustainable fashion through several phases. was facilitated by an injection of $375,000 occasions, gave up his own time to We wish Janet all the best in her retirement. over three years from the GWRDC and contribute so strongly to our bid as an otherwise funded by savings as well as a large industry representative. We must never forget that our endeavor to commercial loan, the latter on track to be The conduct and pursuit of the MNRF bid remain innovative is dependent on our paid back during the financial year 2001/2002. was a heartening exercise where institutions capacity to recruit and further develop cooperated with good-will and a minimum talented individuals within all aspects of the Notwithstanding our past Institute expansions of fuss to generate an outcome not possible wine industry. The Institute is committed to and the construction of the Hickinbotham to achieve by any agent alone. Clearly, we partake in this undertaking with the wine Roseworthy Wine Science Laboratory by have a challenge to put in place a industry as our partner and joint success as Adelaide University, more paint is needed to reasonable MNRF management structure the outcome. complete the picture, chiefly due to the need which is effective and user-friendly, but we to accommodate staff and an increasing are confident this will happen using number of overseas visitors as well as structures which have been proven over the acquiring new equipment for grape and long term. In parallel with our quest to wine research. The Institute, with very strong secure a better funding base for our support from CSIRO Plant Industry, therefore, research, we must also identify costs (and acted as a proponent agent in what later those potentially hidden) associated with turned out to be a successful bid to the various models for its distribution, Federal Government for funding of $4.5 management and execution. Throughout Peter Høj million under the new and once off ‘Major Australian research management, there Director National Research Facility Schemes’ (MNRF). appears to be a case for identifying This will lead to a $7,000,000 upgrade of excellence, fund it appropriately and then let facilities at both the Waite Campus of the the individuals get on with the job within a University of Adelaide and, to a lesser framework defined robustly by major extent, at the Ron Potter Centre of Charles stakeholders, in our case the GWRDC, the Sturt University. Throughout the bid process, Australian wine industry and the we enjoyed strong support from the South communities benefiting from its success.

On a more positive note, the Institute, in addition to the outcomes cited above, again looks back on a year with achievements in a diverse range of areas including our trendsetting NIRS work, the protein based fingerprinting of grape varieties and the application of our unique tannin mouthfeel wheel to the desciption of the mouthfeel properties of chemically defined tannins. Only time will tell if any of these developments rate against the impressive work of past illustrious Institute employees such as Fornachon.

Present and past achievements would not have been possible without the strong support of dedicated wine industry individuals, staff and collaborators. The Institute is most appreciative of this much needed support and is looking forward to a long partnership in our quest to serve the Australian wine industry’s future increased

5 Staff

Peter Bordier Høj, MSc, PhD, Bianca van Wegen, BEnvSc(Hons) Flinders, Jodi-Ann Slade, Library Assistant Copenhagen, Director Part-time Research Assistant (concluded 20/2/01) (commenced 21/11/00) Anna Capone, Library Assistant Wieslawa Cynkar, BSc PhD Wroclaw, (commenced 20/2/01) Research team members Technical Officer *Several members of the Industry Services Gayle Ann Baldock, BSc(Hons) Guelph, Team lead or take part in a number of Paul Anthony Henschke, BSc(Hons) PhD Technical Officer/Casual Analyst research projects. UAdel., Principal Research Microbiologist Jennifer Bellon, Part-time Laboratory Assistant Mark Aidan Sefton, BSc(Hons) PhD UWA, Jane Melissa McCarthy, AdCertMedLabSc Principal Research Chemist USA, CertVetNurs, CertAnimHand TAFE, Analytical Service Elizabeth Joy Waters, BSc PhD UAdel., Technical Assistant (commenced 21 Principal Research Biochemist November 2000). Donald Robert Buick, BSc UAdel., Grad Ian Leigh Francis, BSc(Hons) Monash, PhD Shauna Liam Brown, BBiotech(Hons) Dip(BusAdmin) USA, AIFST, Manager – UAdel., Senior Research Chemist Flinders, Postgraduate Student Analytical Service Robert George Dambergs, BSc(Hons) Anthony John Heinrich, BBiotech(Hons) John Benjamin Hughes, DipWineMrktg UAdel., PhD UQld, Senior Research Flinders, Postgraduate Student UAdel., Analytical Service Supervisor – Chemist (commenced 6 February 2001) Anita Oberholster, BSc(Hons), Stellenbosch, Administration Markus Johannes Herderich, PhD Postgraduate Student (19/4/01) Matthew Grant Holdstock, BSc Flinders, UWuertzberg, Senior Research Chemist Kate Susan Howell, BSc(Hons) UNSW, Analytical Service Supervisor – Laboratory (commenced 9/4/01) Postgraduate Student Gregory Andrew Ruediger, BAppSc SAIT, Yoji Hayasaka, DipEng(IndChem) Tokyo I.T., Kerry Leigh Wilkinson, BSc(Hons) Flinders, GradDipOenol, UAdel., Trace Analysis MPharm Vic. Col. Pharm., CertIntBusMgt Postgraduate Student Laboratory Supervisor Monash, Manager—Mass Spectrometry Facility Carolyn Jane Puglisi, BSc Flinders, Kevin Herbert Pardon, AssDip(AppChem) Miguel Antonio de Barros Lopes, BSc BSc(Hons) UAdel., Postgraduate Student SAIT, Analyst Oregon, PhD UC (Santa Barbara), Agnieszka Janusz, BSc(Hons) Flinders, Andrea Dale Kemp, AssDip(Farm Mgmt), Molecular Biologist Postgraduate Student (commenced 26/3/01) Roseworthy, BA, UAdel., Casual Analyst Eveline Jutta Charlotte Bartowsky, Heather Eunice Smyth (nee Pain), Randell Leith Taylor, BSc(Hons) UAdel., BSc(Hons) PhD UAdel., Research Microbiologist BSc(Hons) UAdel., Postgraduate Student Casual Analyst Zhong Kui Peng, BSc MSc, Zhejiang, PhD (commenced 2/4/01) David Rolfe Boehm, BSc UAdel., Casual Analyst UAdel., Research Chemist Nadia D’Incecco, BAgSc(Food Tech)(Hons) Amanda Louise Cook, AdvCert (Lab Tech) Alan Percy Pollnitz, BSc(Hons), PhD UAdel., Padova, Visiting Italian Postgraduate Mackay, Casual Laboratory Technician Research Chemist Student (concluded 6/2/01) Radka Kolouch, AssDip(Food Tech) Czech Gordon Michael Elsey, BSc(Hons), PhD, Paolo Sivilotti, BAgSc Udine, Visiting Italian Republic, Casual Laboratory Technician Flinders, Postdoctoral Research Fellow Postgraduate Student (commenced Matthew James Cream, Casual Michael Brian Esler, BSc(Hons) USydney, PhD 8/11/00, concluded 31/5/01) Laboratory Technician UWollongong, Postdoctoral Research Fellow Teresa Girbau Solà, BPharm, DipOphtham Jeremy Crispin Hack, Casual George Kyriakos Skouroumounis, Barcelona, Visiting Spanish Postgraduate Laboratory Technician BSc(Hons), Flinders, PhD UAdel., Student (commenced 14/2/01, concluded Bao Tran, AssocDipChemTech USA, Casual Postdoctoral Research Fellow 3/8/01) Laboratory Technician Stéphane Vidal, Eng Dip. (Biochem), Nat. Sandra Hoffman, Visiting German Sandra Lloyd-Davies, BA Flinders, Casual Instit. App. Sc. Lyon, MBiochem, UClaude Postgraduate Student (commenced 18/6/01) Laboratory Administration Assistant Bernard Lyon, PhD UJoseph Fourier Grenoble, Postdoctoral Research Fellow Kenneth Frank Pocock, BAppSc UAdel., Industry Services team members* Administration FAIFST, Senior Chemist Michael Raunkjaer, MSc Copenhagen, Visiting Peter William Godden, BAppSc (Wine Janet Currie Currie, BA Glasgow, MAICD, Research Chemist (concluded 1/12/00) Science) UAdel., Winemaker/Manager Company Secretary (retired 15/1/01) Anders Håkansson, MSc Copenhagen, Industry Services Hans Engelbert Muhlack, BEc Adelaide, Visiting Research Chemist (commenced Mark Gishen, BE(Chem)(Hons) MEngSc(Chem) CPA Aust., Company Secretary Dr Encarnacion Gomez Plaza, Chem, PhD UMelb., Quality Liaison Manager (commenced 3/10/00) Murcia Visiting Research Chemist (two Sally-Jean Bell, BSc(Hons) UWA, GradDip(Wine) Sita Soetratma, B.Bus(Mgmt) USA, month stay, commenced 23/10/00) Roseworthy, PhD UWA, Viticulturist Accountant (concluded 8/12/00) Tracey Ellen Siebert, BSc UAdel., Chemist Adrian Dermott Coulter, BSc Flinders, Rachel Lee Edwards, Accountant (commenced 14/3/01) GradDipOenol, UAdel., Oenologist (commenced 13/12/00) Peter James Costello, BSc (Hons) MSc Peter Valente, BSc(Hons) Flinders, Chemist Heather Margaret Donnell, Secretary to UNSW, PhD UAdel., Microbiologist Ella Margaret Clare Robinson, BA, the Director Holger Gockowiak, BSc(Hons) UAdel., BSc(Hons) UAdel., Chemist Carolyn Debra Grant, AssDip(Acc) Microbiologist (concluded 13/11/00) Part time Raelene Joan Blair, Panorama TAFE, Secretary to the Company Laboratory Manager (commenced 16/4/01) CertAppMgt(Marketing) AIM, Secretary/Conference Secretariat Jeffrey Mark Eglinton, BSc(Hons) UAdel., Communication and Publicity Narelle Elizabeth D’Costa, Microbiologist/Senior Computer Manager/Personal Assistant to the Director Administration Officer Systems Officer Creina Standish Stockley, BSc(Hons) Emma-Kate Darling, Receptionist Dimitra Capone, AssDip(Chem) USA, UAdel., MSc Flinders, MBA USA, Health Maria Concettina Mills, Casual Receptionist Laboratory Technician and Regulatory Information Manager Elizabeth Morgan, Administration Support Maria Jolanta Kwiatkowski, MSc Gliwice, Catherine Grace Daniel, BA ANU, Branka Zepina, Cleaner (commenced 12/3/01) Laboratory Technician GradDip(Lib) RMIT, Librarian Simon Dillon, BSc(Hons) Flinders, Research Ingrid Betty-Maud Oats, DipLibInfo Adel. Assistant (commenced 11/9/00) Tafe, Library Technician

6 Highlights of the year

The outcomes of the Institute’s major Successful organic synthesis of a high- The 14th Advanced Wine Assessment wine closure trial were analysed 20 yielding precursor to the ubiquitous Course was held in July 2000, giving months post-bottling and submitted to wine flavor compund ß-damascenone another 30 participants the opportunity the Australian Journal of Grape and Wine was achieved as a first step to understand to develop and test their sensory Research for publication with expected its formation from non odoriferous evaluation performance. This was the publication of the 42 page article in mid grape precursors. second Course presented under a four- July 2001. day format, which includes over 40 hours of activities over the four days and a Organic synthesis disproved the structural contribution by more than ten senior Flavour scalping studies showed a very large assignment of an oak lactone precursor Australian wine show judges. differential in the rate of disappearance which has been assumed for more than of known wine aroma compounds when 20 years. stored in ‘bag in box’ containers. Institute staff responded to more than 4,400 enquiries in relation to wine Our Analytical Service increased its industry matters. The terminology developed by our sensory turnover and recorded an increase in panel and collaborators to characterise contract research as well as a very large the mouthfeel of red wines is gaining demand for analyses for 4-ethylphenol, The human capital of the Institute is further acceptance as a ‘mouthfeel wheel’ by an indicator of Brettanomyces activity enhanced by qualification of one additional researchers and winemakers worldwide. which may be more widespread than staff member for the PhD degree, by initially assumed. qualification of one additional staff member for the Postgraduate Diploma in Oenology, Gram quantities of nine different tannin by recruitment of a distinguished German fractions from grape seeds and grape A double modification of a laboratory scientist to lead our tannin project and by skins for sensory evaluation were isolated yeast by means of molecular biology the recruitment of a PhD qualified chemist and characterised using HPLC and gives a remarkable demonstration of the with almost two decades of experience in electrospray mass spectrometry followed power of the technology and it is the wine industry. by sensorial assessment by a tasting demonstrated that the genomes of panel over an eight week period. commercial wine strains are quite different to those of laboratory strains.

Combined laboratory and industry-based trials, coupled with chemical Formal ‘Roadshow’ visits were made to and quantitative sensory descriptive SA and Vic in November and December analysis, continued to demonstrate the 2000 (see Appendix 1). Senior Institute potential of S. bayanus, Candida and staff presented seven full-day seminars, new hybrid yeast as positive modulators each seminar consisting of a minimum of of wine sensory characteristics. twelve presentations focussing on current areas of Institute research, or topical issues in the wine industry. A laboratory model system for predicting the effects of fermentation yeast on malolactic bacteria has been developed Institute staff authored/co-authored 37 to enable selection of the most appropriate publications; gave 35 seminars/talks combination of strains to provide greater (excluding the Roadshow seminars reliability of . above); presented 11 workshops; presented 3 posters; gave 38 lectures and supervised 36 students during the year. The Institute secured strong private financial support from Lallemand to boost our research on the effect of wine Institute staff hosted more than 113 yeast on phenolic compounds. international visitors to the Institute during the year including five visiting scientists on extended stay. The Institute paid its final instalment of its $250,000 donation to the Hickinbotham Roseworthy Wine Science Laboratory and The new website of The Australian Wine conducted a series of triplicated ferments Research Institute @ www.awri.com.au on a commercial scale to study the effect was launched including an extensive of winemaking practices and yeast strains educational/troubleshooting section on on colour, mouthfeel, flavour and aroma wine hazes and deposits. of both white and red wines.

Requests for information from the John Fornachon Memorial Library staff increased by 108% over requests received in 1999/00.

7 Staff Activities

In addition to undertaking research and other Creina Stockley is a member of the Senator Judith Troeth, Parliamentary projects described in this report, the Institute Australian Wine and Brandy Corporation’s Secretary to the Minister for Agriculture, performs a large number of external activities Legislative Review Committee, a member of Fisheries and Forestry, Australia (25 in support of the Australian wine industry. the Technical Committee of the Winemakers’ August 2000) Federation of Australia, a member of the Michael Alder, Agriculture, Fisheries and Information on seminars, talks and poster National Reference Committee – Forestry, Australia (25 August 2000) papers given to outside organizations, Environment Strategy Development (South David Monck, South Australian Centre academic lectures delivered, graduate Australian Wine and Brandy Industry for Manufacturing, Marilyn Arnold, students supervised, and the papers published Association) and was elected Vice-President Flavour South Australia, The Food is tabulated and can be found in Appendices of the Nutrition and Wine Expert Group for Industry Association Inc. (28 August 2000) 1–4 of the Annual Report. Activities in addition the Australian OIV Congress. Sandy Grieve, Australian Council of to those in the Appendices are described below. Agricultural Journalists Inc Elizabeth Waters is Leader of Program 1 of the (4 September 2000) Peter Høj is a member of the following: Cooperative Research Centre for Viticulture II. Jenny Eichler and four Year 11 Chemistry Prime Minister’s Science, Engineering and Students from Murrayville College Innovation Council Mark Sefton is on the editorial review (6 September 2000) South Australian Innovation, Science and board of the International Journal of Vine Members of the Executive Committee of the Technology Council and Wine Sciences and is the project leader Australian Academy of Technological Sciences International Trade and Technical Advisory of project 1.3 of the Cooperative Research and Engineering (6 September 2000) Committee (AWBC) Centre for Viticulture II. Ross Dobinson and Tim Hoban, Compliance and Technical Advisory Technology Structuring Limited Committee (AWBC) Leigh Francis is an Editorial Board member (7 September 2000) Technical Committee (Winemakers’ of the Journal of the Science of Food and David Hall and Dr Deann Glenn, Grape Federation of Australia) Agriculture, Editorial Review Board member and Wine Research and Development Wine Committee (Royal Agricultural and of the American Journal of Enology and Corporation (on several occasions) Horticultural Society of South Australia) Viticulture, Chairman of the Royal Australian Richard Heinemann, The Business Centre Waite Campus Management Committee Chemical Institute (SA Branch), Medicinal (14 September 2000) Committee of Management, Viticultural and Agricultural Chemistry Group. H-Michael Schwandt, Consul-General, Publishing, publisher of Australian Journal Federal Republic of Germany, Pat Vaughan, of Grape and Wine Research Ken Pocock serves on the Waite Campus Commonwealth visit co-ordinator, Editorial board of the Journal International Occupational Health and Safety Department of the Prime Minister and des Sciences de la Vigne et du Vin Coordinating Committee. Cabinet, Nektarios Tsirbas, Policy Officer, Conference Planning Committee of the Department of Agriculture, Fisheries and Eleventh Australian Wine Industry Paul Henschke serves on the Editorial Forestry, Thomas Kruckemeyer, Technical Conference (7-11 October Review Board of the following journals: Interpreter, SA, Dean Dempsey, Deputy 2001, Adelaide) (Chair) Australian Journal of Grape and Wine Manager, Protocol, Department of the Australian Organising Committee of the Research; American Journal of Enology and Premier and Cabinet, Peter Hayes, 26th World Congress and 81st General Viticulture; and Australia and New Zealand Rosemount Estates (3 November, 2000) Assembly of the Office International de la Wine Industry Journal. Juan Bournas, Trade Commissioner of Vigne et du Vin (OIV) (11-18 October Chile and Kees van Haasteren, AgTour 2001, Adelaide) and is Section President Peter Godden is a member of the (15 November 2000) of the Oenology Sub-Commission for the Conference Planning and Program sub- Noel Hayes, Amorim (20 November 2000) Australian OIV Congress. Committees and is the Workshop Olga Kostic, International Marketing Coordinator of a program of 72 workshops Manager and Brian Dolling, Managing He is also the Institute’s representative on to be held at the Eleventh Australian Wine Director, NuKorc (22 November 2000) the following Adelaide University’s Industry Technical Conference. Peter Flinn, Agriculture Victoria (22 Boards/Committees: November 2000) Faculty Board Matthew Holdstock is a member of the Tom Angove, Angove’s Wines Department of Horticulture, Viticulture Interwinery Analysis Group Inc. (2 January 2001) and Oenology Advisory Committee Andrew McPherson, Andrew Dean, Hamish Management Committee of the MacGowan, Leigh Clarnette, Guida Vazzoler, Department of Horticulture, Viticulture Visitors to the Institute McPherson Wines; Alister Purbrick, Allan and Oenology Australia George, Neil Larson, Chateau Tahbilk; Eric Parnis and Neil Grant, South Geoff Merrill, Scott Heidrich, Joe DiFabio, Hans Muhlack is the Public Officer of the Australian Department of Industry and Geoff Merrill Wines (2 February 2001) Australian Wine Industry Technical Trade (1 August 2000) Tony Royal, Seguin Moreau Conference Inc. Linda Lee, Hong Kong Trade Development (12 February 2001) Council, Sydney (2 August 2000) Michael Burleigh, Foss Pacific Rae Blair is a member of the Conference Matthew Wingfield, International South (28 February 2001 and 24 April 2001) Planning Committee and is the Treasurer and Australia (2 August 2000) Cecil Camilleri, Yalumba Wines; Alex Sas, Conference Manager of the Eleventh Dr Erika Winter, Agriculture Victoria (3 BRL Hardy; Wendy Allan, Prudence Australian Wine Industry Technical August 2000 and other occasions) Honner, Southcorp Wines; Peter Hayes, Conference. She is also a member of the Group of 30 Advanced Viticulture students Rosemount Estate; Prue Henschke, C.A. Australian Organising Committee of the from South Eastern TAFE, accompanied Henschke & Co; David Bruer, Temple 26th World Congress and 81st General by Denis Vice (3 August 2000) Bruer; Russell Johnstone, Orlando Assembly of the OIV. Linda Bowes and Brian Smedley, SAWBIA Wyndham Group; Vic Patrick, Syd Kyloh, (18 August 2000, and other occasions) Blass Beringer; John Sabine; Andrew Benger; Lou Cremasco; Geoff Hardy,

8 Pertaringa ; James Altmann, Echeverria P. De V; Claudio Salgado India Fruit Doctors; Nic Fazekas, IAMA; Bill Villablanca, Sergio Salgado Segura, Dr K.K. Jindal, Dr YS Parmar University of Murray, IHD, Leon Redunz, AvCare, Armando Fuenzalida Salgado, Sociedad Horticulture and Forestry, Nauni, Solan, Bayer; Keith Jones, South Australian Agricola Chequen Ltda, Ana Maria Flores India (3 August 2000) Wine and Brandy Industry Association; Lopez, Sociedad Agricola Ganadera y DeAnn Glenn, Grape and Wine Research Forestal Arboleda Ltda; Santiago Achurra Israel and Development Corporation; Matt Hernandez, Sociedad Agricola Requingua Amos Mizrach, Volcani Institute, Bet Cahill, Dow AgroSciences; Stephen Ltda, Javier Eduardo Ramirez Vodnizza, Dagan, Israel (23 February 2001) Strachan, Winemakers’ Federation of Sociedad Eduardo Ramirez Letelier y Cia Australia; David Baker, David Braybrook, Ltda; Luis Antonio Paredes Nuñes, Italy Cooperative Research Centre for Sociedad Nuñez y Cia Ltda; Fernando Elio Novello, Gianpalo Vaona, Bolla, Italy Viticulture; Robyn van Heeswijck, Anu Almeda Ollé, Sociedad Vinicola Miguel (2 February 2001) Kuma, Department of Horticulture, Torres S.A.; Gonzalo Bertelsen Diaz, Viticulture and Oenology; Trevor Wicks, Terramater S.A.; José Antonio Esturillo Japan South Australian Research and Castaño, Vinos del Sur S.A.; Dario Polloni Dr Yoshio Kodama, Meiji Seika Ltd, Japan Development Institute; Ary Hoffman, Sch, Viña Cantera S.A.; Roberto I. (15 December 2000) LaTrobe University; Peter Merriman, Echeverria Z, Viña Echeverria Ltda; Alison MacGregor, Rob Sward, Mark Eugenia del Carmen Diaz Aedo, Viña Portugal Krstic, M. Mollah, Department of Natural Morandé S.A.; Teresa Medina Fernández, Antonio Amorim and Professor Miguel Resources and Energy; Rai Kookana, Ray Viña Santa Mónica Ltda; Max Undurraga, Chabral, Amorim, Portugal Correll, CSIRO (27 February 2001) Viña Undurraga S.A.; Jorge Rojas Garcés, (20 November 2000) Natasha Mitchell, ABC radio Viña ValdiviesoS.A.; Eugenio Maffei Rojo, (12 March 2001) Viñedos y Bodegas Corpora S.A.; Angel United Kingdom Imre Lele, Bottle Magic and Ken Craig, SA Domper Pons, Viñedos y Bodegas Dr Steven Walker, Campden & Chorley Centre for Manufacturing (13 March 2001) Domper S.A.; Sara Cabrera Gajardo, wood Food Association Group, United Duncan MacDonald, Rod Pemberthy, Fontec; Tonchi Tomic Jakaf; Carmen Sutil Kingdom (31 August 2000) Sanjay Sinha, Burns Philp (14 March 2001) Servoin; Manuel Madrid Baeza, Mr John Gumbley, Senior Producer, Sky A/Professor Graham Bell, Director, Corporación Chilena del Vino; Chile (15 News, United Kingdom (14 October 2000) University of New South Wales, Centre for November 2000) Eddie Deaville, ADAS Nutritional Sciences ChemoSensory Research (15 March 2001) Research Unit, United Kingdom Rob Lewis, South Australian Research and Denmark (22 November 2000) Development Institute and Nigel Scott CSIRO Thorvald Akselsen, Foss Electric, Carmel Kilcline, Sainsbury’s, United Division of Plant Industry (26 March 2001) Denmark (26 April 2001) Kingdom (3 April 2001) Dr John Radcliffe, CSIRO (27 May 2001) Mai Nygaard, Chr. Hansen, Denmark Professor Ian R Peterson , Coventry Jennifer Gardiner, Adelaide University (17 November 2000) University, United Kingdom (22 May, 2001) (27 May 2001) Libby Boschen, WINETAC (25 May 2001) France United States of America B. Sara, C. Belperio, T. Mitani, K. Mounir Saoma, Michel Picard, France Dr Charles Edwards, Washington State Ringwood, M. Wong, L. Pattison, R. (2 February 2001) University, USA (17 November 2000) Elfenbein, M. Trewartha, Food Laurent Lebrun, Seguin Moreau, France Paul Dolan, Dennis Martin, John White, Technology students from the Regency (12 February 2001) Bob Blue, Nancy Walker, Heath Dolan, Bill Institute of TAFE (31 May 2001) Marc Sabate, Michel De Tapon, Sabaté Leair, Cara Hane, Jill Jepson, Fetzer Vineyards, Robert Druscetta and Anne Patterson, Diosos, France (12 February 2001) Hopland, CA, USA (2 February 2001) Australian Wine and Brandy Corporation Rick Boyer, Marc Imbert, Jekel Vineyards, (28 June 2001) Germany USA (2 February 2001) Don Woolman and Paul Clancy, His Excellency, Mr Karl-Heinz Funke, Paul Ahvenainen, Korbel, USA Australian Wine Industry Suppliers’ Federal Minister of Food, Agriculture and (2 February 2001) Association (29 June 2001) Forestries, Mr Jürgen Detken, Head of Lois Levitan, Cornell University, Cooperative Research Centre for Viticulture Division, Generlal EU Policy, Mr Hellmut Environmental Risk Analysis Program Board members: G. Scollary, D. Gall, D. Altpeter, Director, Economic Relations to Centre for the Environment, USA (27 McWilliam, P. Barnes, T. Martin, J. Dal Bro, non-European Industrial Countries, Ms February 2001) P. Kriedemann, P. Mansfield (4 July 2001) Sigrun Neuwerth, Head of the Press Members of the University of California Professor Edwina Cornish, Adelaide Office, Mr Wolfgang Jaeger, Executive Cooperative Extension Service: Rhonda University (5 July 2001) Officer, Minister’s Office, Ms Ines Mann, Smith – Sonoma, Larry Bettiga – Interpreter, Germany (3 November 2000) Monterey, George Leavitt – Madera, Chile Steve Vasquez – Fresno, Christian Butzke Agapito Garcia Vidal; Ximena Torti Solar, Greece – Enologist, UC Davis, Chuck Ingles – Agricola San Diego de Puquillay Ltda; Yanis Vogiatzis, John Boutari & Son – Sacramento, Jennifer Hashim – Kern, Ed Guillermo Achurra Montes, Agricola S.A., Greece (12 March 2001) Weber – Napa, Paul Verdegaal – San Tunquelen Ltda; Jorge Luis Ollé Cordero, Joaquin, USA (12 March 2001) Agricola y Comercial Crucesillas Ltda; Hong Kong Paul Heck, Scott Laboratories, USA Carlos Valenzuela Olave; Patricio Azocar Antonio Y.L. Koon, Ponti Trading Limited, (27 March 2001) Monasterio, Cooperativa Agricola Kowloon, Hong Kong (2 August 2000) Pisquera Elqui Ltda; Carlos Alfredo Terres Johnny Wan, Hong Kong Trade Other Silva, Cooperativa Agricola Vitivinicola de Development Council, Wanchai, Hong Thirty participants from the International Curico Ltda; Diego Garcia de la Huerta, Kong (2 August 2000) Federation of Agricultural Journalists’ Ninth Juan Sutil Servoin; Maria Adriana Cerda World Congress (4 September 2000) Arriagada, Nevada Export S.A.; Roberto

9 Photograph courtesy of The State Library South Australia Sevenhill Winery: 1900

Sample changer for autotitrator Team Reports

Wine grape tannin and colour specification outcomes from this project are continuously of pigmented material incorporated into being communicated in peer-reviewed polymeric proanthocyanins increases with Staff: Dr Markus Herderich, Dr Elizabeth journals. In this reporting period, three wine age. Waters, Dr Stéphane Vidal, Anita papers have been published, four have been • At present, these analytical protocols are Oberholster, Dr Z Peng, Stella Kassara, submitted for publication, and at least a adopted and validated as routine methods Mariola Kwiatkowski, Dr Mark Sefton, further four are in draft form. Just as for the determination of oligomeric and Dr Leigh Francis, Yoji Hayasaka, Gayle important is the public and international polymeric polyphenols in grape, seeds, and Baldock, Dr George Skouroumounis, recognition of the achievements obtained by wine samples from numerous viticultural and Heather Donnell, Richard Gawel the Tannin project. A fine example is the winemaking trials. Libraries summarizing Collaborators: Adelaide University - terminology developed by our sensory panel UV- and mass spectral data will facilitate Dr James Kennedy, Dr Robert to characterise the mouthfeel of red wines, the identification of key polyphenols in Asenstorfer, David Lee, Stephanie which - after publication in 2000 - instantly the future. These initiatives will result in Lambert, Dr Graham Jones, Renate was accepted and identified as ‘mouthfeel significantly reduced analysis time and Ristic, Dr Patrick Iland, Stephen Clarke wheel’ by researchers and winemakers all secure higher through-put of samples. around the world. The international This project was initiated as collaboration significance of the outcomes of the Tannin Wine pigments between The Australian Wine Research project is also reflected by various invited Robert Asenstorfer’s PhD thesis was passed. Institute and The University of Adelaide at the presentations, for example, the Institute was • The findings from Robert Asenstorfer’s request of industry; an Industry Reference invited to give a 45 minute plenary thesis have now been prepared for Group composed of twelve senior winemakers presentation entitled ‘Red wine colour and publication by Robert and Dr Graham Jones. advises it. Funding for the tannin project is mouthfeel: Analysis of phenolic pigments provided by the GWRDC as well as by the CRCV. and tannins’ at the Annual Meeting of the In a collaboration with Robert, Yoji Hayasaka American Society for Enology and Viticulture, developed and optimised methods for the The ambitious objectives of this challenging held in San Diego from 27-30 June 2001. detection of stable wine pigments by multi-disciplinary project are: This task was admirably performed by the Electrospray ionastion mass spectrometry • To gain an understanding of the types of new project leader and appeared to (ESI-MS/MS). Applying tandem mass compounds important to red wine colour generate much interest and follow-up spectrometric experiments numerous and mouthfeel and of conditions enquiries from parties interested in complex red pigments could be detected in favouring their presence in wine. establishing collaborations. wine samples. With this approach we also • To determine whether the interaction of identified four novel anthocyanin polyphenol proanthocyanidins (tannins) with other A similar reception was given when the tannin derivatives for the first time as natural grape and wine compounds modifies their project and some of its recent outcomes products in wine. sensory properties. were presented by Dr Vidal at an international • To establish viticultural and oenological conference in Bordeaux, where much praise A paper outlining this methodology has techniques that enhance wine colour and was given for our achievements in a difficult been submitted for publication. desirable mouthfeel. area of research. More specific achievements relate to outcomes listed below. Analytical methods to identify the existence General observations of new wine compounds is a necessary but The new Leader of the Tannin Project, Dr Method development not sufficient tool to advise winemakers on Markus Herderich, commenced at the Institute Methods to quantify the levels of tannins in how to modulate wine composition to meet on 9 April 2001. Dr Jim Kennedy, post- seed extracts and in wine have been developed, set quality specifications. Once identified, doctoral fellow at the University of Adelaide, and the identity of polymeric proanthocyanidins the relative importance of such compounds has accepted a tenure track position as an (tannins) has been rigorously demonstrated must be established. The next step is thus Assistant Professor in the department of by mass spectrometric analysis. to synthesize some of the newly discovered Food Science and Technology at Oregon The results have been published (Z. Peng, compounds in sufficient quantities to State University and left the project in May Y. Hayasaka, P.G. Iland, M. Sefton, P. Høj describe their chemical and sensorial 2001. Other fluctuations of the involved and E.J. Waters, J. Agric. Food Chem., 49, properties. To this end, David Lee, a PhD staff include Robert Asenstorfer, student at 2001, Institute publication #660) and student funded by CRCV, and Mr Anders the University of Adelaide, who qualified for another manuscript was submitted for Haakansson, a visiting synthetic chemist the award of the degree of Doctor of publication (Z. Peng, P.G. Iland, A. from Denmark, has initiated synthesis of Philosophy; Anita Oberholster, PhD student Oberholster, M.A. Sefton and E.J. Waters). these new wine pigments. at the AWRI, who finished the experimental The main outcomes are: part of her PhD research; and Stella Kassara, Copigmentation chemist at the AWRI, who left for maternity • Reversed phase HPLC methods have been Copigmentation reactions are strongly leave in July 2000 and recently resigned. developed for quantitative analysis of influenced by pH, with its relative importance polymeric procyanidins (tannins) in grape being greatest at higher pH values. Ethanol Despite these changes good progress has seed extracts and for quantitative analysis decreases the copigmentation interaction. been made and the appointment of Dr of polymeric pigments in wines. The Markus Herderich as new leader of the recovery of polymeric pigments with this Of the common simple phenolics in wine, Tannin project has further focussed to our method was 95%. quercetin has been shown to form the most activities. Dr Liz Waters, the preceding stable intermolecular complex with malvidin- project leader, is thanked for her significant • Results obtained with the HPLC method 3-glucose. past contribution which, in the longer term, correlated well with spectrophotometric was not sustainable due to her demanding measures of total pigmented polymers Self-association of malvidin-3 glucose also task as Manager of Program 1 of the and indices of wine age. appears to be a major cause of colour Cooperative Research Centre for Viticulture. • Using this technology, a study of 120 enhancement. As model experiments show Besides other measures of success and wines has confirmed that the proportion that the extent of intermolecular copigmentation progress, it is very pleasing to see that the depends on the concentration of anthocyanins,

11 Team Reports

more is likely better in young red wines. Shiraz grapes were collected during the Institute, Stéphane Vidal commenced work Conversely, aged products feature significantly period of berry development from a range of in Véronique Cheynier’s laboratory at INRA- reduced concentrations of anthocyanins and the viticultural regions (Riverland, Barossa and IPV in Montpellier on 6 June 2000, where he contribution of intermolecular copigmentation Coonawarra). In some cases samples were prepared tannin fractions from seeds and to the color of older red wines remains to be collected from trials where different skins for sensory assessment. established. Stephanie Lambert, an Adelaide irrigation schedules had been applied. These University PhD Scholarship holder, presented studies showed that levels of catechin, Stéphane has isolated and purified gram a poster on her physical chemistry studies at epicatechin and extractable tannins quantities of ten different tannin fractions In Vino Analytica Scientia, held in Bordeaux (procyanidins) in seeds increased up until from grape seeds and grape skins. from 14-16 June. , where they were at their maximum, and then declined. The A panel of tasters has been convened and Grape seed and skin proanthocyanidins differences in seed composition of grapes trained by Richard Gawel over the last few As reported previously, adding extra seeds to sourced from the range of viticultural regions months. In late March these tasters a red wine ferment produced wines with were less than the differences in skin colour completed a very extensive tasting of the increased levels of phenolics, including (anthocyanin levels). tannin fractions isolated by Stéphane. tannins and pigmented polymers, and enhanced colour density. When the seed colour of berries from This is the first time that such fractions different vineyards within one region are have been tasted and profiled for their After ageing in bottle for two years, wines compared at the same stage of ripeness astringency sub-qualities. The data will made from must with extra seeds (e.g. 24 ºBrix) there is little difference in allow us to understand how the chemical contained more colour and were higher in the degree of seed colour, indicating that properties of tannins relate to their astringency than wines made once berries have ripened to a particular sensory properties and are currently being conventionally. Importantly, the wines point, assessment of seed colour is not a subjected to extensive statistical analysis. appeared not to be more bitter. discriminating factor between the potential quality of different vineyards. Stéphane presented this work at In Vino This material is being drafted for Analytica Scientia, held in Bordeaux from publication (Z. Peng, P.G. Iland, I.L. Francis, A viticultural trial to investigate the differences 14-16 June. Y. Hayasaka and E.J. Waters). between seed physical characteristics (seed number, size and weight) and seed chemical Winemaking practices that effect A method for characterising the composition composition of grapes that develop in proanthocyanidin levels and sensory of grape and wine tannins using acid shaded canopy conditions versus that of properties of wine hydrolysis in presence of phloroglucinol has grapes that develop in moderately and well been developed. exposed canopy conditions has been To study the effect of winemaking practices established by Pat Iland and Renata Ristic at on colour and mouthfeel the first commercial The analytical protocol has been published The University of Adelaide. scale winemaking trials have been (J.A. Kennedy and G.P Jones, J. Agric. conducted at the Hickinbotham Roseworthy Food Chem., 49, 2001, 1740.). The extractable tannin content of seeds Wine Science Laboratory located on the was maximal in the three weeks prior to Waite Campus with support from our For the first time, nano electrospray veraison and then decreased as the fruit Industry Services Manager and continued ionisation in combination with mass ripened. The seed content of the flavan-3- input by the facility Manager, Mr Stephen spectrometry allowed us to detect ol monomers also decreased after veraison. Clarke, a winemaker with 20 years individual tannins on a molecular level. commercial experience. This method has been developed by Yoji Visually seeds were initially green, plump Hayasaka and represents a major progress and had pliable seed coats. Beginning Winemaking was performed according to the for our studies aiming to understand the from veraison, the seeds browned in guidelines of the industry reference group. chemical properties of tannins. colour, became desiccated and the seed coats hardened. The objectives of these initial 20 Application of this methodology enabled fermentations are to analyze the influence us to characterize the composition and These changes in content and visual of red grape varieties (Coonawarra Shiraz purity of several different tannin fractions aspects can be explained by oxidative and Adelaide Hills ), from grape seeds and grape skins. This is events occurring in the seed coat during fermentation tanks (rotary tanks versus essential when attempting to ascribe fruit ripening. This information has been stationary fermenters), and fermentation specific sensorial properties to individual published (J.A. Kennedy, G.J. Troup, J.R. temperatures (18°C, 25°C) on red wine classes of isolated tannins. Pilbrow, D.R. Hutton, D. Hewitt, C.R. phenolic compounds. A complementary Hunter, R. Ristic, P.G. Iland and G.P. Jones, trial conducted by our Wine Microbiology An investigation on structural changes of Aust. J. Grape Wine Research 6, 2000, 244). team assessed the influence of yeast grape skin tannins and pigmented strains on tannin profiles. polymers at different stages of berry There were no differences in total development has been conducted. Again, amounts (monomers and procyanidins), In addition, we aim to study the variation the methods developed proved to be subunit composition (amount of terminal between replicate fermentations and to essential for this study on the composition and extension units) or mean degree of identify key parameters which contribute of grape skin proanthocyanins. polymerisation, of seeds of different to variation. canopy treatments. The results of the latter experiments have Reference samples of juice and skins have been submitted for publication (J.A. Linking the astringency and mouthfeel been collected on a daily basis during Kennedy, Y. Hayasaka, S. Vidal, E.J. Waters properties of wines to proanthocyanidin fermentations, and are continuously kept and G.P. Jones – manuscript in review). and or other polymer fractions in wine on a weekly basis prior to bottling. Funded by the CRCV and employed by the

12 Comprehensive chemical analysis of several hundred samples for polyphenol and tannin content has been initiated. It will be complemented with sensory analysis after short term bottle storage, and several of the wines produced will be presented to industry in workshops conducted at the Eleventh Australian Wine Industry Technical Conference.

An analysis of compositional differences between grape marc derived tannin material and tannins extracted from skins during fermentation, indicates differences in the relative proportions of polymeric vs monomeric pigments and the ratio of malvidin-3-glucoside-6’ acetates to malvidin-e-glucoside-6’-p-coumarate anthocyanins. For example, the proportion of these natural polymeric pigment in young wines is approximately 15% but can constitute greater than 60% in grape marc Dr Leigh Francis and Dr Bob Dambergs extracts. Similarly, the proportion of malvidin-3-glucoside-6’-p-coumarate is higher These matters have now been largely Australian calibration or set of in marc extracts than in most young wines. resolved and the potential for calibrations. The significance of regional, commercialisation of the technology has varietal and especially seasonal variation These characteristics can be determined been examined more fully. While it was for NIR analysis will be further investigated. by a single HPLC analysis using the deemed timely to delay publication earlier to At this point in time, it appears that region- methods developed by Peng. allow the Institute to secure the advantage specific and variety-specific calibrations of being an early entrant into the field by will be slightly better than pan-Australian Further analysis of a liquid grape marc not disclosing information to potential calibrations. Whether the differences have extract indicated that dextrins were not commercial competitors or imitators, it is real practical consequences is yet to be detectable in the preparations analysed. suggested that this delay is no longer resolved (see Figure 1). This contrasts with previously published warranted. The publication of the NIR method results on the composition of solid grape for colour, pH and total soluble solids, will Negotiations with the CRCV and the marc extracts. now be accelerated. collaborating wine companies have taken place in order to clarify the mode of The sample collection program provided commercialisation and provide a mandate Rapid instrumental techniques more than 2000 samples from the 2001 for the management of such activity. vintage from grape growing regions in Staff: Dr Leigh Francis, Dr Bob Dambergs, Dr NSW, VIC, WA as well as SA. It is The transfer of calibrations between well- Michael Esler, Dr Bruce Kambouris, Dr anticipated that by the end of 2001 the matched or standardised instruments Wies Cynkar, David Boehm, Professor chemical and spectroscopic analysis of this appears likely to be successful and will be Peter Høj, Dr Sally-Jean Bell, Peter Godden data set will be completed and the data examined in detail with samples from the Collaborators: Adelaide University - used to refine earlier NIR calibrations 2001 vintage. It should be noted Dr Graham Jones, Dr Patrick Iland based on 1999 and 2000 data and however that this is not a trivial exercise Orlando Wyndham Group - Russell develop something approaching a pan- and may require significant effort. While Johnstone, Ben Zander, Inca Lee Southcorp Wines - Dr Andrew Kleinig, )

Eric Wilkes -1 AB 2.0 A comprehensive suite of accurate and precise calibrations has been developed for the measurement of colour, pH, and TSS in red 1.5 grapes on three distinctly different types of NIR instruments. These calibrations range in 1.0 specificity from general 'pan-Australian' to more precise region and variety specific. N = 909 samples N = 78 samples Greater than 2,000 samples have been collected 10 varieties 1 variety (Cab.Sauv.) 0.5 10 regions 1 region (Riverland) this vintage by collaborating companies and R2 = 0.90 R2 = 0.94 some of these have been delivered to the anthocyanins by NIR (mg g Total SECV = +/-0.14 mg g-1 SECV = +/-0.05 mg g-1 Institute and analysis has commenced. 0.5 1.0 1.5 2.0 0.5 1.0 1.5 2.0 As part of our obligations to the Cooperative Total anthocyanins by reference method (mg g-1) Research Centre for Viticulture and to the Commonwealth, publication of this method Figure 1: NIR calibrations for total anthocyanin concentration for a population of ~900 red wine grape samples and one of its sub-populations. A. ‘Global’ calibration; 909 samples across 10 growing regions, 10 grape varieties and 2 seasons, coloured had earlier been held over pending by variety. B. Subpopulation, localised for both a single grape variety, Cabernet Sauvignon, and a single growing region, finalisation of intellectual property issues. the Riverland. Observe that NIR measurement precision (SECV) improves by focusing on a particular region and variety.

13 Team Reports

the direct calibration of a single Disappointingly, there have been unforeseen • Development of new isotopic dilution instrument has proven to be relatively delays and only one instrument is likely to assays for quantifying the key impact straightforward, the simultaneous be supplied to industry in 2001. compounds produced by yeast, as a tool standardisation of a population of several for determining the importance of yeast or many instruments to the one calibration Considerable effort has been expended in strain and fermentation conditions, is a more complex issue. In particular, it investigating previously identified variation in • Evaluation of the oenological properties of appears that not all types of instruments the glucosyl-glucose (G-G) assay calibrations non-conventional yeasts with an emphasis are equally capable of being standardised. (particularly for white grapes). Detailed on correlating chemical composition and On the other hand, there now exists scrutiny of the reference method has led to organoleptic attributes of wines made recognised protocols for the some refinements of the analytical techniques. under winery conditions, standardisation of more expensive In particular, the use of a 'stomacher' for gently • Laboratory and winery studies concerning research grade instruments, such as the macerating white grapes in a reproducible the use of mixed yeast strain starter NIRSystems6500 spectrophotometer. manner that is also more relevant to real cultures, compared with conventional Commercially available software that is winemaking practices, has improved the single strain cultures, as a strategy for designed for calibration transfer and degree of uncertainty in the measurement. simulating 'natural ferments', and instrument standardisation is to be evaluated in standardisation of the Further samples of in particular Quantification of yeast key impact aroma Institute’s NIRSystems6500 spectrometer are being collected this season with compounds—methods development with BRL Hardy’s based on 2000 and cooperation from BRL Hardy. These samples As stated in the previous annual report, the aim 2001 vintage data. will be tracked from juice right through to of this collaboration with Drs Mark Sefton and wine and linked with the subsequent quality Leigh Francis and their teams, is to develop Investigations with instruments capable of rating determined at the in-house allocation stable isotope dilution assay methods for scanning whole fruit are continuing in tastings. Other wines assessed at the quantifying the key impact compounds formed association with two wine companies, but allocation tastings will also be scanned and during the yeast alcoholic fermentation. Work no data has yet been received. The Perten then discrimination models will be tested. has focused on the principal esters produced by DA7000 instrument offers the possibility yeast which are an important contributor to the of being able to rapidly analyse samples of Studies using commercial tasting data fruity/floral/estery character of wine, which is intact whole berries. The investigation is indicate the approach of using NIR for often referred to as the 'fermentation bouquet' proceeding on a large data set already allocation tastings and measurement of G- of wine. Good progress has again been made provided by BRL Hardy using samples from G in white fruit may have some value and with the synthesis of ten isotopically-labelled the 2001 vintage. this will be addressed more fully in future ester standards being completed. The work. Samples from the 2001 vintage development of validated assays for the ethyl Trials with an existing prototype bin probe have now been received at the Institute esters has commenced. Ms Tracey Siebert, who have commenced and data is being collected. and work will commence shortly. has recently joined the team, will focus on Initial trials have focussed on off-site evaluation applying the assays to wines produced through of reproducibility of spectral collection. In some recent preliminary studies, grape and our fermentation trials; a series of Chardonnay other tannin fractions of various sizes, chemical and Cabernet Sauvignon wines are currently Trials with prototype bin probes continued composition and degree of polymerisation undergoing extensive chemical analysis and during the 2001 vintage to further that have been prepared by Dr Stèphane sensory evaluation. This analytical approach will examine the feasibility of the concept. Vidal as part of project AWR 96/1 (Wine allow us to relate the ester profile of wine Whilst the trials have demonstrated the grape tannin and colour specification) have determined chemically and by sensory promise of this approach of in-situ been scanned by NIRS. The spectra appear techniques and to determine the relative scanning, there is some indication that to show significant differences and suggest importance of yeast strain and fermentation commercial development of a NIR ‘bin- that the technique may be useful in conditions on the development of the probe’ may be ambitious at present given distinguishing these tannin fractions either fermentation bouquet of wine. In addition the current state of commercially available quantitatively, qualitatively, or both. to assays on esters, several other stable NIR technology. Institute efforts may be isotope dilution assays for important wine better focused on addressing the more aroma and flavour compounds is underway. tractable issues encountered in Yeast flavour and fermentation activity development of a compact laboratory- Industry evaluation of selected novel based instrument. The latter is likely to be Staff: Dr Paul Henschke, Dr Eveline Bartowsky, flavour yeasts deliverable to industry earlier than the Dimitra Capone, Dr Peter Costello, Nadia Replicated winery scale fermentation trials former. D'Incecco, Jeff Eglinton, Dr Leigh Francis, were undertaken during the 2001 vintage Holger Gockowiak, Kate Howell, Jane with the aim to consolidate the preliminary Three of the relatively-inexpensive McCarthy, Dr Mark Sefton, Tracey trials undertaken previously on a laboratory prototype NIRTech instruments will be Siebert, Professor Peter Høj scale and to focus on the key attributes that tested in parallel at the Institute in order Collaborators: Professor Graham Fleet, these non-conventional yeasts can offer wine to evaluate instrument effects in the The University of New South Wales, makers in search of increasing the flavour absence of all other experimental Stephen Clarke, The University of Adelaide diversity of wines. variables. The successful standardisation of cheaper diode-array based instruments This GWRDC-funded project has as a primary Saccharomyces bayanus trials in particular (such as this prototype), is still objective, to investigate microbiological In addition to the fermentation trials a subject of considerable research inquiry interventions which can be used to generate referred to in the past annual report, a internationally. a wider spectrum of wine sensory properties winery scale trial using S. bayanus, and S. than currently achieved with conventional use cerevisiae as a reference wine yeast, with It was planned to expand the trial of these of well-characterised Saccharomyces cerevisiae Chardonnay and Cabernet Sauvignon has prototype instruments to a small number strains and well-known malolactic bacteria. been undertaken in the Hickinbotham of companies for the 2001 vintage. Three current main activities are related to: Roseworthy Wine Science Laboratory in

14 Photograph courtesy of The State Library South Australia Seppeltsfield : 1945

Atomic absortion spectrometer Team Reports

collaboration with Mr Stephen Clarke during Mechanically harvested Cabernet Sauvignon Candida stellata and Candida krusei trials the 2001 vintage. The Wine Science fruit, which had been treated with SO2 Due to tank limitations in the Hickinbotham Laboratory offers the opportunity to have during , was crushed and inoculated Roseworthy Wine Science Laboratory, 18L the wines contract made essentially under with S. bayanus or S. cerevisiae. Problem- Chardonnay (BRL Hardy) fermentation trials were winery conditions but the close proximity to free ferments with both yeast completed in performed in glass with the experimental the Institute also allows us to closely monitor 13-14 days, however, starter culture-induced Candida strains, C. stellata AWRI 861, 1159 and fermentation progress. We are grateful to MLF was delayed in the wines fermented C. krusei 863, at the Institute. The Chardonnay BRL Hardy for generously providing 4000L of with S. bayanus compared with S. cerevisiae. juice provided by BRL Hardy was also used for Chardonnay juice and 4 tonnes of Cabernet these trials. The ferments were superinoculated Sauvignon grapes for use in these trials. All trial wines are currently being filtered and with S. cerevisiae AWRI 838 at approximately bottled in readiness for extensive chemical, 50% fermentation progress to ensure the The main purpose of the Chardonnay trial instrumental and sensory analysis. In particular, production of wines with low residual sugar. The was to evaluate the performance and to we plan to quantify the main yeast-derived S. cerevisiae AWRI 838 ferment failed to determine the organoleptic properties of S. flavour compounds of these wines with GC-MS complete and was rescued by reinoculation with bayanus AWRI 1375 under larger scale analysis combined with the newly developed AWRI 838. The wines were cold-settled, racked, conditions using 260-280L fermentors. As in previous trials S. cerevisiae AWRI 838 was the reference yeast. In addition, treatments were included which were designed to evaluate different methods for ensuring that wine with low residual sugar was achieved after fermentation with non-conventional yeasts, in this case S. bayanus 1375. The treatments involved:

• Sequential inoculation with S. cerevisiae AWRI 838 when the S. bayanus ferment was approximately 70% completed, • Aerobic pumping-over of a stuck S. bayanus ferment, and • Inoculation of a stuck S. bayanus ferment with an S. bayanus AWRI 1375 rescue culture prepared by the Institute recommended procedure.

The reference wine yeast, S. cerevisiae AWRI 838, completed fermentation at 15°C in 18- 24 days whereas S. bayanus AWRI 1375 ceased fermentation with a residual sugar of 40 g/L after 35 days fermentation. Sequential inoculation of the S. bayanus ferment with S. cerevisiae AWRI 838 prevented lagging of the S. bayanus ferment which completed in 39 days. Both the aerobic pumping over treatment and the use of the S. bayanus rescue culture successfully removed the residual sugar from the stuck S. bayanus ferment. The latter procedure gave Dr Peter Costello and an especially interesting result because the rescue procedure not only worked well with stable isotope dilution assays developed by the treated with SO2, membrane filtered and bottled. the same yeast that caused the stuck team of Dr Mark Sefton and Dr Leigh Francis Formal sensory descriptive evaluation and fermentation but that the Institute's to establish the chemical basis for the flavour chemical analysis has commenced. recommended rescue procedure was attributes of this yeast and the impact of Preliminary analysis confirms our previous successful with the less robust S. bayanus fermentation conditions on their formation. findings (Institute staff publications #614 fermentation yeast compared with the and #625) that the wines made with normally recommended S. cerevisiae. The Several of the winery trial wines will be different yeast have marked differences effect of these treatments on wine chemical selected for use in the Winemaking with especially in the ester profile (floral versus composition and sensory characteristics is non-conventional yeasts workshop being banana versus fruity versus ethyl acetate). being determined as a guide to planned in conjunction with the Eleventh Chemical analysis is recorded in the Table 1. recommending treatment preference. Australian Wine Industry Technical Conference to be held in Adelaide in October 2001. The The chemical and sensory data confirm the Based on encouraging results obtained with team is quietly confident that the soon to be potential of novel yeasts for developing S. bayanus AWRI 1375 in a previous performed rigorous analysis and results will wines with novel attributes. laboratory-scale Shiraz trial, a larger scale point to the AWRI 838 and AWRI 1375 trial using six 800 kg capacity rotary strains giving very different wines with fermentors located in the WSL was differing positive attributes. undertaken during the 2001 vintage. S. cerevisiae AWRI 838 was the reference yeast.

16 Optimisation of C. stellata inoculation for oxidation with H2O2 or aeration, or by nutrient limiting conditions in a Chardonnay inducing fermentation acetaldehyde treatment. The treatment with juice. Wines were also made with each of Some non-Saccharomyces yeasts are known acetaldehyde is experimental and is not the five strains. These treatments resulted in to exhibit greater sensitivity than S. cerevisiae permitted under existing wine regulations. three wines made by mixed culture ferments to growth inhibition by SO2, which is often (M1-M3) and five wines made by monoculture added to grape juice/must prior to inoculation Yeast impact on wine flavour and ferments (T1-T5) under the two nutrient with a selected wine yeast to inhibit indigenous aroma—mixed culture versus conditions. Three blended wines were also yeasts and to prevent browning; this observation monoculture fermentation prepared by mixing in equal proportion the was noted in the Institute staff publication #625 It is universal practice to induce the alcoholic monoculture wines which corresponded to in relation to C. stellata. Since this property fermentation of grape juice/must with a the yeasts used in the mixed culture ferments may impact on the effective deployment of selected wine strain of Saccharomyces (B1-B3). The wines were tested for organoleptic this strain by industry, the sensitivity of C. cerevisiae in order to produce a reliable differences by duo-trio aroma testing. stellata to SO2 has been quantified. fermentation and a wine with expected chemical and organoleptic properties. The proportion of each yeast strain in the The inhibitory affect of SO2 on yeast growth However, a relatively small, but increasing mixed culture ferments was monitored by was determined in a freshly prepared grape proportion of wines are made without microsatellite-PCR analysis. The juice by monitoring the delay in the onset of inoculation thereby allowing the indigenous microsatellite sequences, present in the yeast fermentation. C. stellata 1159 showed an yeasts to proliferate, to give a so-called chromosomal DNA, were demonstrated to extended lag phase of 2-4 days when ‘natural ferment.’ This practice, which some be sufficiently polymorphic to permit reliable inoculated into sultana juice containing winemakers believe gives wines preferred strain identification. free/total SO2 of at least 32/50 mg/L (pH sensory characteristics, can be unreliable and 3.2) whereas S. cerevisiae commenced its application is limited because the wine Duo-trio testing of the resultant wines noticeable growth within one day. By contrast making properties of the indigenous yeasts showed that in some cases multiple strains with C. stellata, C. krusei was more tolerant are not known in advance. of S. cerevisiae interacted to produce wines and commenced good growth with a that were sensorially different to: minimal lag phase. The low tolerance of C. A key difference between inoculated and • Any of the wines produced by stellata to SO2 is likely related to low production non-induced fermentation relates to the monoculture fermentation with the of SO2 and acetaldehyde by this yeast. initial population of S. cerevisiae present. corresponding yeasts, or Non-induced ferments typically initially • A wine produced by blending in equal In addition to the use of H2O2 to reduce the contain a mixture of indigenous non- proportion the three wines produced by SO2-induced growth lag by oxidation of the Saccharomyces yeasts which dominate the the corresponding monoculture SO2, the lag phase in grape juice was also early stage of fermentation but then give fermentations. abolished by the addition of acetaldehyde, way to a mixture of S. cerevisiae strains which presumably acted by binding the toxic which complete the fermentation of the These preliminary results need verification, but free SO2 , even at a concentration of 65 sugars whereas induced ferments, while also nevertheless suggest that wine with an aroma mg/L SO2. Recent research from Prof. having a similar population of indigenous profile different from wines of single yeast Pamment’s laboratory, University of yeast species are generally dominated by the ferments, or their blends, can be produced Melbourne, has shown acetaldehyde to inoculated S. cerevisiae yeast culture from by mixed culture fermentation only when stimulate fermentation independent of the outset. This project was undertaken to the yeast strains interact and no-one strain reaction with SO2 so it is possible that both evaluate the use of a mixed culture strategy dominates, as appears to normally be the mechanisms have stimulated yeast growth. to simulate ‘natural’ ferments and to case for 'natural' ferments. This suggests investigate the interaction between strains. that mixed cultures of S. cerevisiae in wine These data confirmed that C. stellata, like As a simplified model, the mixed culture fermentations could be used to modify the most indigenous yeasts, is more susceptible ferments were performed with three strains reminiscent of natural than S. cerevisiae to SO2 requiring either of S. cerevisiae wine yeast. ferments but with a minimal risk typical of reduced SO2 addition before inoculation, inoculated monoculture fermentations. prolonged storage of juice at low temperature Five different strains of S. cerevisiae in three to ensure maximal binding of the free SO2 different combinations of three strains were or a reduction in free SO2 content by tested under nutrient limiting and non-

Treatment Alcohol Glycerol pH TA VA Succinate free SO2 total SO2 (% v/v) (g/L) (g/L) (g/L) (g/L) (mg/L) (mg/L)

Juice 3.24 7.5 13 35

S. cerevisiae 838 13.5 5.9 3.39 7.5 0.41 0.5 22 112

C. stellata 1159 + Sc 838 13.4 14.8 3.28 9.5 0.94 0.8 24 107

C. stellata 861 + Sc 838 13.4 10.0 3.39 7.6 0.21 1.0 26 131

C. krusei 863 + Sc 838 (13.8) 6.6 3.37 7.3 0.16 0.8 28 102

Range for malate = 2.3-2.5 g/L, lactate <0.1 g/L, tartrate=4.0 g/L, and citrate=0.3-0.4 g/L

Table 1. Comparison of the chemical composition of Chardonnay wine made by fermentation with S. cerevisiae AWRI 838, C. stellata AWRI 861, 1159 and C. krusei 863.

17 Photograph courtesy of The State Library of South Australia

Orbital shaking water bath with small scale fermentations Team Reports

Ms Kate Howell has performed this work at ß-glucosidase activity, which cleaves Completion of MLF ranged 2 - 8 weeks for the Institute as part of her PhD studies glucosides formed by the action of the the four strains in the Chardonnay wine undertaken through the University of NSW previously mentioned endoglycosidase whereas the same four strains completed with supervision by Dr Eveline Bartowsky, enzymes or are directly derived from the MLF within two weeks in the Cabernet Professor Graham Fleet (University of New grape, to release the flavour aglycon. Sauvignon wine, suggesting a nutrient South Wales) and Dr Paul Henschke. limitation or an inhibitory metabolite may Monitoring the total a-a content of the have selectively delayed the growth and Role of malolactic bacterial glycosidase model wine containing the glycosidic extract fermentation activity of two of the strains in enzymes in the development of wine flavour showed that purified ß-glucosidase only the Chardonnay wine. The peak diacetyl Nadia D’Incecco, a PhD student from The provided limited hydrolysis of the wine concentrations were lower and of greater University of Padua, Italy, working at the glycosides and confirmed that -L- range in the white wines (1-4.5 mg/L) Institute under the supervision of Dr Eveline arabinofuranosidase and -L-rhamnosidase compared with those obtained with the red Bartowsky and Dr Paul Henschke has completed were required for efficient hydrolysis of the wines (8-10 mg/L). As for the red wines her studies on determining the role of ß- glycosides. MLF conducted by the O. oeni (Figure 2), diacetyl peaked after the glucosidase activity, produced by malolactic strain only led to limited hydrolysis of the consumption of malic acid and when bacteria, on the release of glycosylated wine glycosides, suggesting that this approx. 75% of the citric acid had been flavour precursor compounds isolated from bacterium does not elaborate sufficient metabolised (Figure 2). After the complete Chardonnay wine. This study complements activity of the enzymes necessary for the removal of citric acid, diacetyl had fallen to her work in Italy where she is characterising hydrolysis of disaccharides. Addition of -L- less than 50% of the peak value. the ß-glucosidase activities of yeast. arabinofuranosidase and -L-rhamnosidase to the wine during MLF demonstrated that These data agree with our bottled wine survey The role of malolactic bacteria in the these enzymes are required for efficient that red wines carry a higher diacetyl development of wine flavour is not clear but hydrolysis of grape glycosides during MLF. concentration than whites (Institute staff is believed to involve at least three publication #546). These data also showed a mechanisms, i) deacidification, by which L- This work has provided important new correlation between peak diacetyl concentration malic acid is decarboxylated to L-lactic acid information on the somewhat uncertain role and duration of MLF, although further work and CO2, ii) metabolism of wine substrates of MLF in wine flavour development. This is needed to confirm this observation. The to form either flavoured or flavourless investigation, which requires confirmation kinetic pattern of malic and citric acid products, such as the metabolism of citric with additional strains, suggests that the degradation and diacetyl evolution was acid to form diacetyl with a 'buttery' flavour, ability of ML bacteria to release glycosylated qualitatively similar for all strains in each wine. acetic acid, and 2,3-butanediol which has no flavour precursor molecules is relatively flavour significance in wine (reviewed by restricted and probably confined to the ß- In conclusion, these results suggest that Bartowsky and Henschke 2000, staff reprint glucosides, unless yeast derived or extraneous although strains vary in ability to produce #637), and iii) the hypothesised release of glycosidases, such as rhamnosidases and diacetyl, other factors, such as duration of grape derived glycosylated flavour precursor arabinofuranosidases, are present. MLF, and the stage of MLF at which the compounds by excreted glycosidic enzymes wine is stabilised (SO2 addition or membrane (GWRDC project UCS 92/4). filtration) appear to give relatively greater Malolactic fermentation and wine flavour differences in diacetyl concentration. The An experiment was designed and conducted choice of ML bacterial strain is important to to establish whether or not malolactic Staff: Dr Paul Henschke, Dr Eveline the extent that growth response in a particular bacteria have the ability to hydrolyse and Bartowsky, Dr Peter Costello, Holger wine is variable, thereby significantly release glycosylated flavour precursor Gockwiak, Jane McCarthy influencing the final diacetyl concentration. compounds isolated from Chardonnay wine. Collaborators: Dr Leigh Francis, The flavour precursor compounds present in AWRI, and Dr Andrew Markides, Most importantly, these data confirm the wine are typically ß-glucosides or Adelaide University result obtained previously with red wine that diglycosides in which an arabinose, diacetyl is a transient metabolite and that rhamnose, apiose or xylose residue is linked Sensory attributes associated with the the timing of wine stabilisation (e.g. sulfite to the inner glucose residue (Institute malolactic fermentation of wine - diacetyl addition) for controlling the desired level of publication #458). An extract of glycosides formation by commercial MLF bacteria in wine diacetyl in wine is important. was made from a Chardonnay wine, added The flavour compound, diacetyl, is a major to a model wine, and after MLF, the glucose contributor to the ‘buttery’ attribute of wine Preliminary evaluation of a chemical sensor content of the wine was measured by the which has undergone malolactic fermentation with wines having undergone MLF glycosyl-glucose (GG) assay to indicate the (MLF). The ML bacterial strain can affect the A small project has been undertaken to evaluate extent of hydrolysis and a GC-MS analysis diacetyl content of wine (Institute publications the ability of a chemical sensor to determine performed to reveal the extent of liberated #546 and #637), however, limited comparative the aroma profile of wines which had flavour aglycons. Control experiments information on commercially available strains undergone MLF. The chemical sensor included no bacterial addition to test for is available to winemakers. This project aims to instrument, a Hewlett Packard model HP4440, acidic hydrolysis, and addition of various systematically characterise these strains under located in the Department of HVO of combinations of glycosidic enzymes as similar conditions in a red and Adelaide University, is a mass spectrometry reference positive controls. (alcohol 12.5-13.0% v/v, malic acid 2.5 g/L, based ‘electronic nose’ for the citric acid 0.5 g/L, pH 3.4, SO2 negligible). The characterisation of wine volatiles, where the Using synthetic substrates, the two strains of present report relates to diacetyl production by fragmentation patterns may provide a Oenococcus oeni studied were shown to four ML bacteria cultures, Chr. Hansen Viniflora fingerprint of the volatile compounds present exhibit low -L-arabinofuranosidase and -L- oenos, Lallemand EQ54, Lallemand OSU, and in the headspace of the wine sample. This rhamnosidase activities, endoglycosidases, Condimenta/Lallemand Bitec/Viacell proVino, in instrument comprises an autosampler (capacity which are necessary to cleave the outer a 2000 Chardonnay wine, the results pertaining for 44 samples) and a mass sensor where sugar, often arabinose or rhamnose, from to the red wine (1998 Cabernet Sauvignon) trial the wine volatiles are ionised and fragments the diglycoside. These strains had significant were summarised in the previous annual report. detected, and the abundance of ions

19 Team Reports

This work has demonstrated that the 10 1.0 recently defined model grape juice medium can support yeast growth and completion CFU/mL 8 0.8 of alcoholic fermentation and the model malic acid wine supports MLF and completion in a 6 0.6 diacetyl timely fashion which is reflective of a ‘real’ grape juice/wine. The model also 4 0.4 L - lactic acid reflected grape juice studies with respect * citric acid to the effect of timing of inoculation with & L-lactic acid (g/L) 0.2 Diacetyl (mg/L), Cell 2 * ML bacteria on the time course of MLF. viability(cfu/mL), Malic * Acetic & Citric acid (g/L) * acetic acid 0 0.0 * * * * * A test system for predicting the stimulately 0 5 10 15 20 25 30 and inhibitory affects of wine yeast on Days after inoculation malolactic bacteria

Figure 2. Relationship between diacetyl and organic acid metabolism by Oenococcus oeni in Cabernet Sauvignon wine As stated previously, the aim of this project is to develop a test system for detecting determined. The instrument has a major on the factors which allow the wines to stimulatory and inhibitory interactions advantage over conventional instruments in be distinguished. However, this sensor between wine microorganisms, and to use that it can rapidly analyse many samples. cannot detect diacetyl, the main this system to characterise the interaction compound conferring the 'buttery' between pairs of popular wine yeast and ML Two wine types (Chardonnay and Cabernet character, in the presence of alcohol. bacteria so that winemakers can choose the Sauvignon) from three different viticultural most appropriate combination for their wine regions (Chardonnay – Eden Valley, Sunraysia Model studies on the effect of timing of process. Previously, we have described the and Padthaway; Cabernet Sauvignon – Eden O. oeni inoculation on the induction of MLF growth response of one Oenococcus oeni Valley, Coonawarra and Swan Hill) were Based on the chemically defined grape strain to four fermentative yeast strains using subjected to MLF with two commercial bacteria juice/wine medium reported on in the 1999 the model test system. Because this test preparations (Viniflora oenos and Lalvin Annual Report, a trial has been undertaken system uses a chemically defined medium to MCW). Malolactic fermentation was carried to evaluate this medium for the ability to reproducibly simulate the important out at 20ºC in an oxygen-free atmosphere. model aspects of the MLF. This investigation nutritional and chemical properties of grape focused on the effect of timing of O. oeni juice, work is in progress to validate this test The chemical sensor could distinguish inoculation on the initiation and duration of system with several different grape juices. between both the red and white wines, and MLF and diacetyl production. A compatible pre and post MLF wines, and in some cases yeast (Lalvin CY-3079) and ML bacteria strain Juice prepared from hand-picked, disease free, was also able to distinguish the wines on the (AWRI Lc5p) were chosen. The ML bacteria undamaged grapes has been ameliorated to basis of origin, possibly on the basis of were inoculated at different stages during closely match the chemical composition of the fermentation esters. There was evidence alcoholic fermentation (AF) (mid exponential model medium, with respect to sugar, acidity, that the bacterial strain used to conduct the yeast phase and at approximately 5 ºBrix nitrogen and so on. Wines have been produced MLF could be distinguished. However, the sugar), immediate post AF and 8 weeks post from one juice with the test yeasts and the chemical sensor was unable to distinguish AF, in the presence or absence of yeast, to growth response of the test ML bacterium wines on the basis of the flavour compound, examine the effect on MLF duration and the has been determined. Quantitatively, the diacetyl, due to the diacetyl ion fragments effect of yeast on diacetyl content. growth response was lower in terms of overlapping those of ethanol. Further work biomass yields when compared with the is required to establish the identity of the It was found that the timing of O. oeni model medium, but qualitatively similar volatile compounds which allowed for the inoculation affected the initiation and results were obtained, indicating that the differentiation of wines pre and post MLF duration of the MLF as reported by others test system acts as a grape juice model, with a particular bacteria strain. To for wines. MLF was found to initiate and based on these results derived from a single unequivocally assess the use of the chemical complete more rapidly when O. oeni was juice. Following validation with the additional sensor for discriminating wines from inoculated towards the end of the stationary grape juices, the model system will be used different regions, further work would phase and during the decline phase of yeast to screen various combinations of popular required, for example, standardised wine growth, particularly if the yeast was removed commercial wine yeasts and ML bacteria. making of several examples of fruit from before inoculation. When O. oeni was several viticultural regions. inoculated in the presence of fermenting Aspects of this work have recently been yeast, the MLF was delayed until near the communicated to industry via i) Institute This project was undertaken by Ms Helen completion of the alcoholic fermentation. Roadshows to winemaker groups in South McCarthy as an Honours project in the Insufficient diacetyl data have been obtained Australia and North-western Victoria during Department of Horiculture, Viticulture and to date to show any clear association with November/December 2000, ii) presentation to Oenology, with supervision by Drs Eveline inoculation for MLF and diacetyl content. the Department of Horticulture, Viticulture and Bartowsky and Leigh Francis. Helen was Oenology Grape and Wine Industry Outreach awarded a first class honours degree, and This project was undertaken by Ms Marie Seminar, University of Adelaide, November commenced as Graduate Winemaker with Pearce as an Honours project in the 2000, and iii) ‘Can interactions between wine Southcorp Wines at Karadoc in December 2000. Department of Horticulture, Viticulture and yeast and lactic acid bacteria be more Oenology, with supervision by Drs Eveline predictable?’ formed part of an article called This work has provided an insight into the Bartowsky and Andrew Markides, The Clearer answers sought for old questions potential use of the chemical sensor as a University of Adelaide. Marie was awarded which appeared in the Aust. N.Z. Wine Ind. means to analyse wines, and may prove a first class honours degree, and commenced J., Nov./Dec. 2000. This project was useful as a quick method for the initial as Graduate Winemaker with Southcorp undertaken as a collaboration with Dr examination of wines for MLF character, Wines at Coonawarra in January 2001. Andrew Markides of The University of and the technique could give information Adelaide (GWRDC Project UA 92/3).

20 20 The model test system shows promise for This study was undertaken by Ms Nina The suitability of Near Infrared Spectroscopy the relatively rapid laboratory assessment Viergutz as an Honours project in the for measurement of yeast assimilable of the inhibitory, neutral or stimulatory Department of Horticulture, Viticulture nitrogen content of grape juice affect of wine yeasts on the growth and and Oenology, with supervision by Drs As reported previously, this project, which is fermentation of a ML bacterial strain based Peter Costello and Andrew Markides, The being pursued in collaboration with GWRDC on the results obtained with a single grape University of Adelaide. Nina has been AWR 98/2, has the objective to develop and juice sample. Validation of the test system awarded an honours degree, and evaluate near infrared spectroscopy (NIRS) with additional juices is in progress. commenced as an Assistant Winemaker for estimating the yeast assimilable nitrogen with Andrew Peace Wines, Victoria, in content (YAN) of grape juice as measured by Preliminary characterisation of an MLF early February 2001. established reference chemical methods. stimulatory fraction from wine The poor correlations previously observed Based on recent reports from the literature These results suggest that stimulation of between NIRS and chemical measures of that a yeast mannoprotein fraction could be malolactic bacterial growth may be related YAN were thought to relate to low signal stimulatory for MLF, a small project was more to yeast produced growth factors, such due to the low relative concentration of free undertaken to investigate this hypothesis as vitamins, amino acids and peptides than amino acids in grape juice. Work on using a chemically defined grape juice to the macromolecular fraction. Our future evaluating several techniques to enhance the medium to simplify analysis. Wines were work will aim to characterise which factors, weak NIRS signals has recommenced with Dr made by fermentation with AWRI 838, an if any, are responsible for MLF stimulation. Bob Dambergs recently joining the Institute's MLF stimulatory yeast, and AWRI 839, a Such knowledge could be used to manager NIRS project team. yeast with neutral affect on MLF. The wines malolactic fermentation better. were clarified by filtration, and contained <5 Restarting stuck ferments mg/L total SO2. Various factions of these This subproject, detailed in previous annual wines were prepared with 30,000 and 3,000 Microbiological analysis of industry reports, has been concluded with a molecular weight cut-off ultrafilter technical problems communication to industry (Eglinton and membranes. The effect of these fractions on Staff: Dr Paul Henschke, Dr Eveline Henschke [2001] Aust. N.Z. Wine Ind. J. 16: the growth of a test strain of O. oeni has Bartowsky, Dr Peter Costello, Jeff 77–81. Institute staff publication #657). been assessed by supplementation of a Eglinton, Holger Gockowiak, Jane This paper relates to the effect of acetic acid synthetic wine medium. McCarthy, Dr Leigh Francis, Mark Gishen, concentration on the ability to restart a stuck Dr Michael Esler, Dr Bob Dambergs fermentation. Aspects of this work with Growth and malic acid degradation was flavour implications have been taken up in greatest in the unfractionated wine, In addition to investigating specific winery project GWRDC AWR 2. A S. bayanus followed by the 3K eluate, 3K retentate, microbiological problems this project also Chardonnay ferment which stuck in the 30K retentate, and synthetic wine medium. provides resources for maintaining the Hickinbotham Roseworthy Wine Science This result suggested that the low molecular Institute's culture collection which provides Laboratory was successfully restarted with a weight material was most stimulatory to yeast and bacteria for teaching and research, rescue culture of S. bayanus, prepared by the bacterial growth, more so than the high performing advanced wine microbiological Institute's recommended method, indicating molecular weight fraction, known to contain analysis, undertaking dried yeast quality that yeast other than certain strains of S. (manno)proteins. HPLC analysis of the testing, and providing microbiological cerevisiae can be used to restart stuck fractions revealed that the yeast-derived consultation to industry personnel. A ferments under appropriate conditions. This protein content was highest in the workshop on the identification and wine, and others rescued by aeration/stirring unfractionated wine and the 30K retentate. oenological characteristics of wine spoilage or with S. cerevisiae are undergoing sensory With the exception of the unfractionated microorganisms is being planned for the evaluation to determine the effect of the wine, the protein content of the fractions 11th Australian Wine Industry Technical rescue procedure on wine flavour. gave a negative relationship to bacterial Conference in October 2001. growth and malolactic activity. Mousy off-flavour occurrence and formation in wine Two manuscripts communicating the microbiology of mousy off-flavour formation Yeast 1 in wine to industry have been published: 60 Stimulatory • Grbin and Henschke (2000) Mousy off- Yeast 2 } flavour production in grape juice and wine by Dekkera and Brettanomyces yeasts. Aust. J. Grape Wine Res. 6: 255–262 40 (Institute publication #650); • Costello, Lee and Henschke (2001) Ability of lactic acid bacteria to produce N- heterocycles causing mousy off-flavour in 20 Yeast 3 wine. Aust. J. Grape Wine Res. (in press). Neutral Reference } The main points of these papers are that all Culture turbidity (NTU) 0 Yeast 4 Inhibitory species of Dekkera/Brettanomyces yeast and many species of the heterofermentative 0 5 10 15 lactic acid bacteria, including some strains of Time (days) the wine malolactic bacterium Oenococcus oeni, are capable of producing mousy off- flavour N-heterocyclic compounds under Figure 3. Growth of Oenococcus oeni in model wines made by fermentation with four different yeast strains and the unfermented reference model wine. Relative to bacterial growth in the model wine, yeast 1 and yeast 2 are designated to be certain conditions. Aspects of stimulatory, yeast 3 neutral and yeast 4 inhibitory. Brettanomyces/Dekkera management in

21 Team Reports

winemaking in relation to mousy off-flavour excretion); extracellular metabolism strains showed a similar ranking for each and formation of volatile phenols was (enzymatic); and production of chemically yeast. One yeast consistently demonstrated presented to the Southern and physically reactive metabolites (such as, an ability to allow the production of wines Workshop held in Methven, Canterbury, carbonyls and polysaccharides). Many of with low colour density whereas another New Zealand, 18-21 January 2001. these reactions and their implications for consistently enabled the elaboration of wine properties are not well researched, and wines with high colour density. These results more specifically at the applied level, the suggest that yeast strains have a significant Yeast interaction with red grape specific properties of individual wine strains influence on the phenolic content of wines. phenolics and effect on wine are not well characterised. Studies are currently in progress to better sensory properties understand the nature of this observation A laboratory scale fermentation procedure with the view to make recommendations Staff: Dr Eveline Bartowsky, Simon Dillon, using replicated quantities of red grape must about the suitability of individual yeasts for Dr Paul Henschke (0.25-1.0 kg) has been shown to give specified fermentation outcomes. statistically reproducible fermentation A collaboration between the Institute and profiles and wine chemical composition. Lallemand has been established to investigate Wine colour density measurements were also Selection and improvement of wine the interactions between yeast and grape found to reproduce well. yeasts by application of molecular biology phenolics during red wine fermentation. Comparison of 17 commercial wine yeast for Staff: Dr Miguel de Barros Lopes, Dr Paul Lallemand is contributing a substantial sum their ability to effect extraction and or retention Henschke, Jeff Eglinton, Anthony of money for our investigations, for which of various grape phenolic compounds into Heinrich, Jenny Bellon and Professor we are most appreciative. wine was examined using triplicated 1 kg Peter Høj lots of Shiraz grapes (Clare Valley). The Red grape must contains a variety of phenolic grape must was adjusted to pH 3.5, 25 ºBrix In the past few years a number of molecular compounds which contribute to the colour, soluble solids, 250 mg/L YAN and 25 mg/L genetic projects studying different aspects of aroma, flavour, texture and stability of wine. SO2, and fermentation was conducted at wine yeast have been carried out, mostly The principal groups of phenolic compounds 25ºC with no protection from the air during within the program of the previous CRCV-1. are the monomeric (flavanols, proanthocyanidins, cap plunging. All 17 yeast strains completed These included studies on hydrogen sulfide anthocyanidins and flavonols) and polymeric alcoholic fermentation within 23 days. On production, killer yeasts, nitrogen utilisation, phenolics (tannins). Fermentation yeasts are the basis of wine colour density analysis, the acetic acid formation and glycerol production. believed to interact with many of these yeast strains were ranked into three groups Many of these projects have been concluded, compounds in a complex manner, involving which generally reflects industry and the students researching these areas physical, chemical and enzymatic mechanisms. observations, as reported by Lallemand. have recently completed their thesis. During These interactions may include: liberation of the past year Chris Smyl received his PhD phenolics by chemical/enzymatic Colour density comparisons of wines made and is currently doing post-doctoral research of grape tissues; loss from wine by with grapes obtained from three viticultural on tannins at The University of Adelaide. adsorption to the yeast cell wall; intracellular regions (Clare Valley, Langhorne Creek and Nicholas Yap also received his PhD and is metabolism (uptake, enzymatic modification, Adelaide Hills) and fermented with six yeast presently employed in the Australian wine industry. Two of the projects, acetic acid formation and glycerol production, have been combined in order to successfully produce a yeast that accumulates less ethanol during fermentation.

Construction of a wine strain producing less ethanol In previous annual reports a commercial wine yeast overexpressing the glycerol synthesis gene (GPD2) has been described. These yeasts (GPD2-OP) accumulate two to three fold more glycerol than non-modified strains during grape juice fermentation, and divert sugar away from ethanol production. These modified strains, however, also increase the acetic acid concentration to amounts generally unacceptable for winemaking (Institute publication #649). Several strategies have been tested to decrease the amount of acetic acid. The strategy that appears to be the most successful is the modification of a second gene ALD6, which encodes for an aldehyde dehydrogenase. Deletion of the ALD6 gene (ald6∆) in an otherwise wild-type strain decreased the amount of acetic acid more than three-fold, demonstrating that this is an effective strategy of decreasing acetic accumulation during fermentation. Importantly, the concentration of acetic acid produced Simon Dillon and Dr Eveline Bartowsky

22 when the cells overexpress the GPD2 gene was also diminished when the ALD6 gene was deleted (from 1.42 g/L to 0.36 g/L). In fact, the accumulation of acetic acid with the strain modified at both genes (GPD2-OP / ald6∆) was lower than that of a non-modified strain (0.66 g/L). The biosynthesis of glycerol remained elevated (increasing from 6.0 g/L to 16.3 g/L), with a simultaneous decrease in the concentration of ethanol (from 31.1 g/L to 27.3) (Note: these ferments were done with an initial sugar concentration of 80 g/L).

Gas chromatography/mass spectrometry (GC/MS) was used to demonstrate that GPD2/ALD6 modification affected the biosynthesis of a number of secondary metabolites of fermentation, including acids, esters, aldehydes and higher alcohols, many of which are flavour-active. The concentration of acetaldehyde, acetoin and 2,3-butanediol are the compounds most affected by the increase in GPD2 expression. Also, the concentration of a number of ethylidene glycerols (acetals of glycerol and acetaldehyde), Jeff Eglinton, Dr Miguel de Barros Lopes and Alan Pollnitz that have been previously found in other commercial wine strains and other species cerevisiae parent. These results demonstrate wines that contain high concentrations of with winemaking potential are being made. that producing hybrids between S. cerevisiae acetaldehyde such as , increased more Hybrids between strains of S. cerevisiae and wine strains and other species is a useful than ten-fold in a GPD2 overproducing strain. other Saccharomyces species have been strategy for acquiring yeasts with novel The formation of secondary metabolites was successful. These hybrids generally share winemaking characteristics from species that influenced by the inclusion of oxygen, which properties of both parents, including ethanol are less suited for grape juice fermentation. also affected the growth rate, final cell number, tolerance and the ability to grow in media One of the main advantages of this method and sugar consumption of the culture. The containing high levels of sugar from the wine is that it is considered safe and not classified results indicate that modification of GPD2 yeast, and improved growth at low as genetic modification. and ALD6 expression represents an effective temperature from the second parent. A strategy to increase glycerol concentration commercial wine yeast X S. paradoxus hybrid Identifying genes for wine yeast and decrease ethanol and acetic acid has been used in a winemaking trial. The hybrid improvement concentrations in wine. These modifications fermented grape juice at the same rate as the Saccharomyces cerevisiae is a model biological alter the chemical composition of the wine commercial wine yeast parent, even though organism and the first eukaryote to have its such that, potentially, novel flavour diversity the S. paradoxus parent was unable to ferment chromosomal DNA completely sequenced. This is possible. A double modification of a yeast grape juice at all. Results indicate that the has been followed by a systematic approach by means of molecular biology gives a genome of these hybrid yeasts is stable during to study the function of each of its more remarkable demonstration of the power of fermentation. Chemical analysis demonstrates than 6000 genes. The strains used in these the technology. A paper describing some of that the composition of the wine is changed studies are laboratory strains of S. cerevisiae, this work has been published (M. de Barros when the hybrid yeast is used. For example, which have important physiological differences Lopes, A.U. Rehman, H. Gockowiak, A.J. a significant increase in the glycerol when compared to commercial wine strains. Heinrich, P. Langridge, P.A. Henschke. concentration, without a corresponding For example, phenotypic studies have confirmed Australian Journal of Grape and Wine increase in the acetic acid concentration, that wine strains tolerate an increased sugar Research, 2000, Institute publication #649). was seen in the wine made with the hybrid and alcohol concentration, and generally A second manuscript describing the more yeast. Further chemical studies and sensory multiply more rapidly. Also, wine yeast produce recent work has been submitted for publication. analysis are necessary to evaluate the secondary metabolites during fermentation winemaking properties of the strain. It is also that are positive for wine flavour. The gene Use of non-molecular techniques for the being tested if this hybrid yeast ferments and regulatory sequences that specify these improvement of wine strains more efficiently at low temperature, a key wine yeast properties are unknown. A In order to produce novel wine yeasts with characteristic derived from the non-S. DNA approach (amplified fragment length unique characteristics, hybrids between polymorphism - AFLP) has been used to identify genes that differ between wine and Compound GPD2 GPD2-OP GPD2 GPD2-OP laboratory strains. These results demonstrate ALD6 ALD6 ald6∆ ald6∆ that the genome of commercial wine strains are quite different to those of laboratory Glucose (g/L)1 3.3 4.25 10.95 2.9 strains with several genes duplicated and novel sequences present. Glycerol (g/L)1 5.1 13.4 6.0 16.3

Ethanol (g/L)1 34.1 26.0 31.1 27.3 To further understand the mechanisms behind the phenotypic differences between Acetic acid (g/L)1 0.66 1.42 0.20 0.36 yeasts, and hence to understand what makes a wine yeast perform well under Table 2. Concentration of some metabolites for wild-type and modified strains different fermentation conditions, two-

23 Team Reports

dimensional gel electrophoresis (2D gels) has Grape composition and wine flavour The program is divided into three broad areas: been employed. This system allows proteome the identification of grape-derived wine comparisons of different yeast strains under Staff: Dr Mark Sefton, Dr Leigh Francis, Yoji components that affect wine aroma and flavour; different growth conditions. The wine and Hayasaka, Stella Kassara, Gayle studies of the formation and degradation of laboratory strains display distinct protein maps, Baldock, Professor Peter Høj, Wies these wine flavour constituents including the which supports the AFLP results showing that Cynkar, Dr George Skouroumounis, identification of their grape-derived precursors; there are important differences between the Kevin Pardon, Dimitra Capone, Dr and, ultimately, studies on how viticultural two types of strains. Initially, this technique is Markus Herderich, Dr Gordon Elsey, variables affect the production of wine flavour being used to identify proteins that may provide Carolyn Puglisi compounds and their precursors in the grape. improved ethanol tolerance of commercial In order to achieve these aims, it is necessary strains. The results show that the expression This project has the long-term objective of to have available routine, sensitive and accurate of several proteins are increased in response to understanding how the combination of analytical methods for determining the ethanol stress. A subset of proteins also appear viticultural and winemaking practices concentration of these grape and wine to be destabilised by ethanol. Proteins of interest determines the aroma and flavour components in various matrices. The are being isolated from the gel, digested with characteristics of wine. While some aroma development of such methods is, and continues specific proteases, and the resulting peptides and flavour compounds present in finished to be, a high priority of the program. analysed by matrix-assisted laser ionisation/ wine are derived unchanged from the grape, desorption time-of-flight spectrometry the majority are formed by biological and Black pepper aroma in Shiraz wines (MALDI-TOF) in collaboration with Dr Graeme chemical processes from flavourless grape Earlier work on this project had shown that Currie at the University of Melbourne. The metabolites. To achieve the aims of this the black pepper character can be tasted proteins can then be identified by comparison project thus requires not only an and scored in Shiraz grapes from the 1999 to databases that exist for the well studied understanding of the compounds that are vintage. Analytical data had indicated that laboratory yeast. Characterising proteins that responsible for wine flavour but also the certain compounds, that are reported to be are altered during wine fermentation will be processes by which they are formed. Only involved in the aroma of peppercorns, occur beneficial for tracking wine fermentations, then can viticultural practices be linked to in higher concentration in the high-pepper predicting strain attributes and selecting and flavour outcomes in finished wine. Shiraz samples, than in the low-pepper producing new improved strains. Shiraz samples.

Further grape lots were obtained from vineyards identified from historical data as producing wines with peppery characteristics. Fruit from the 2000 season from four separate vineyards in Coonawarra, and two lots of grapes from a central Victorian , have been studied. Following informal tasting of the berry samples, only one lot of fruit (from the Central Victoria vineyard) was characterised as high in pepper flavour, and this plus a low pepper sample has been analysed by GC/MS. From data collected from the 1999 samples analysed, it was intended to confirm whether the compounds identified previously were also present in the 2000 season samples, and carry out more detailed investigations. The project is now on hold as a result of the key researcher, Ms Stella Kassara, having taken maternity leave and subsequently resigned. This project will resume once a suitable replacement for Stella is recruited. With the consent of the GWRDC, salary savings from this project were employed to further boost the Institute’s NIR-research by securing the services of Dr Bob Dambergs.

Synthesis of potential wine flavour compounds formed from grape-derived glycosides In the past, we have tentatively identified several compounds (four megastigmatrienones and four maleimides) in mild-acid hydrolysates of grape-derived glycosides. These mild-acid hydrolysates have been demonstrated, by formal sensory studies, to confer honey, lime and woody characteristics to base wines to which they have been added (Institute publication #420). Enzyme hydrolysates Kevin Pardon, Dr Mark Sefton and Carolyn Puglisi

24 Photograph courtesy of The State Library South Australia Nicholas Brooks at ‘Brookside’: 1930

GC/MS with tray samples Team Reports

which do not confer these characteristics, employed towards the synthesis of a glycosidic The influence of oak cooperage on wine do not contain these compounds. They are derivative of this acetylenic precursor. The composition patented as flavour additives for other reactivity of this glucoside at wine pH will products, but are not commercially available. then be determined. Staff: Dr Mark Sefton, Dr Alan Pollnitz, In order to assess the impact of the individual Kevin Pardon compounds on wine flavour it has been Analysis of other wine flavour compounds necessary to produce them by synthesis. and their grape-derived precursors Several experiments on shelf-life and Precise, sensitive and relatively simple extraction rates have been conducted with The four megastigmatrienones has been analytical methods, employing stable commercial sized French and American oak successfully prepared, while the synthesis of isotope-labelled internal standards, are chips, and also with fine oak shavings. No the four substituted maleimides is well continuing to be developed for quantifying significant change in amounts of extractable advanced. Following some difficulties with known key grape-derived wine flavour oak lactones, vanillin, guaiacol or 4- the final step of the first two sequences that compounds. These methods once completed methylguaiacol were observed after six were attempted, an alternative route was will enable future research on viticultural and months storage of the commercial-sized successful, and two of the target oenological controls of flavour formation, chips in plastic bags. compounds were obtained. These and will be added to over time with further compounds will be prepared in sufficient compounds identified from the literature or Extraction of volatile oak components quantities for sensory investigation. Progress from on-going research. Following the from finer shavings was faster than from towards the synthesis of the remaining completion of stability studies of deuterium- chips over the first week of extraction, but target compounds, based on the successful labelled analogues, fully validated methods the amount of volatile compounds strategy towards preparing the first two, is have been developed for measuring ß- extracted over a longer period (28 days) well advanced, and this task should be damascenone (fruity and honey like was similar. Soaking for this longer period completed shortly. A sensory evaluation of character) - and ß-ionone (berry, violets), is necessary to extract most of the flavour all of these compounds will then be isobutylmethoxypyrazine (capsicum/vegetative), from commercial sized chips. undertaken – to ascertain their importance nerol, geraniol and linalool (floral/citrus), for wine sensory characters. The 1,1,6-trimethyl-1,2-dihydronaphthalene Earlier work in the Institute had shown that compounds that are found to have very (TDN, kerosine character in aged the presence of oxygen can influence the strong sensory characteristics at wines) and cis-rose oxide (lychee character, effects of heating on oakwood composition. concentrations typically found in wine will be particularly in Traminer wines). Lignin breakdown by heat is more extensive the target of future viticultural and Deuterium-labelled ß-phenylethanol, which when oxygen is present and this results in oenological experimentation. has a floral/rose-like aroma, and labelled ß- greater amounts of vanillin, guaiacol, and 4- phenylethyl acetate (fruity) have been methylguaiacol as well as cis- and trans-oak Precursors to damascenone successfully prepared. lactone being formed in the oak compared ß-Damascenone is one of the most to when oak is heated in an inert important flavour compounds known to Two grape-derived precursors to the atmosphere. This indicated that the science; it is found in many fruit and important wine aroma compound known as composition of heated shavings, which have vegetable products, including wine where it ‘wine lactone,’ which has woody and a high surface area to volume ratio, will be imparts ‘stewed apple’, ‘fruity’ and honey- coconut-like aromas in concentrated form, quite different to that of larger pieces of like characters, and tonne quantities are have been synthesised on a large scale. heated oak, a supposition supported by produced commercially for the perfume and These compounds will be studied to recent experiments. flavouring industries. Recent studies by us determine the rate at which they can be have indicated that ß-damascenone in wine converted to wine lactone under typical When oak which had been chipped or results from the hydrolytic breakdown of bottle storage conditions. A parallel shaved to different sizes was heated in air, complex grape-derived secondary synthesis of deuterium-labelled analogues of the finer fragments (shavings) were richer in metabolites formed from carotenoids such as these compounds has also been completed. extractable volatile constituents than were neoxanthin (Institute publication #630). The deuterium-labelled analogues will be the coarser fragments. The effect of heating used in future viticultural trials for measuring the finer shavings in the absence, compared Two synthetic sequences towards an acetylenic the variability in concentration of these to the presence of air, was similar to that of damascenone precursor were developed in precursors in grapes. heating coarser versus finer shavings. The parallel, and the target compound has now effects of air and chip size on the been obtained. Consequently, we have been Ms Agnieszka Janusz from Flinders University composition of heated oak pieces was not able to identify this precursor as an intermediate has recently taken up an offer of a CRCV the same for all compounds. in the conversion of a grape-derived postgraduate scholarship to work on the norisoprenoid to damascenone in fruit, and identification of new wine flavour compounds Thus the relative proportions as well as a preliminary hydrolytic study in model wine has formed from glycosides derived from Shiraz absolute amounts of oak volatiles was shown that this precursor gives damascenone and Cabernet Sauvignon grapes. Ms Heather affected by these parameters, and in high yield. Once grape samples from this Smyth (ne Pain) has taken up an Institute smoky/toasty aromas are more likely to be year’s vintage have been processed, extracts postgraduate scholarship to study the predominant when smaller pieces of wood of these samples will be examined to determine relationship between wine composition and are heated, especially if heated in air. whether glycoconjugates of this precursor flavour in young Riesling and Chardonnay are components of ripe fruit, and whether it wines. As previously stated, when oak volatiles are will be desireable to develop techniques for totally extracted into a wine from oak shavings measuring these compounds in fruit as and the shavings are then removed, these quality parameters. Further material is being volatiles can continue to be generated prepared for detailed hydrolytic studies at hydrolytically, presumably from precursor forms, room temperature. The more efficient of the also extracted from the oak samples. two synthetic sequences is now being Further studies on exhaustively extracted oak shavings have shown that when these shavings

26 are resuspended in a fresh sample model wine, 4-ethylphenol in red wines – implications berries, must and wine. Such an vanillin can continue to be formed over time. for microbial problems identification technique would complement The formation of vanillin was greater when In previous staff publications (Institute and extend the identification service air was present. This demonstrates that oak publications #624, #627), we pointed out currently available, since DNA fingerprinting contains non-extractable precursors of vanillin as that a survey of Australian red wines is valid for berries and must only. This has well as precursors which can be extracted into indicated a relatively high concentration of been explored and led to a novel approach wine. The accumulation of oak components 4-ethylphenol, a product of Brettanomyces for varietal discrimination of fruits. in wine during barrel ageing is therefore not activity, was found in some Australian red only a function of extraction rates but is also wines at concentrations above 500-600 Proteins with a wide range of masses (13 to 33 dependent on this hydrolytic generation. mg/L. 4-ethylphenol can impart an kDa) were found in the juices of 19 different This new insight has implications for the undesirable ‘medicinal’/ ‘bandaid’ character. varieties of grape and were identified as mostly potential development of ‘oaky’ characters Using our stable isotope dilution assay, our thaumatin-like proteins and chitinases. Small in wine following bottling. research group measured the concentration consistent differences in molecular masses of 4-ethylphenol in wine stored in 44 were noted when otherwise identical proteins Precursors to oak lactone American and 47 French new and used oak were compared from different varieties. These Despite their sensory importance, the origin from several suppliers. Wine stored differences persisted through different harvest of cis- and trans-oak lactone remains in shaved and refired oak barrels contained years and in fruit grown in different locations. unclear. These compounds are already up to 85% less 4-ethylphenol and 4- Based on the definition of 4 different masses present in green oakwood, but additional ethylguaiacol than wine stored in normal for thaumatin-like proteins and 12 different quantities of these compounds can be barrels of the same age that were not masses for the chitinases and using statistical generated in the wood during the drying shaved. The concentration of 4-ethylphenol analysis, the methods developed could be used (seasoning) and coopering processes, in oak found in 61 bottled commercial Australian for varietal differentiation of grapes on the extracts heated to 50ºC, and even in the red wines of various ages ranged from 2 basis of the protein composition of the juice. injector block of a gas chromatograph µg/L in a up to 2660 µg/L in a Shiraz, A refereed paper describing this work has during analysis of oak extracts. These with a mean concentration of 795 µg/L. The been published (Institute publication #663). observations indicate the presence of one or highest level of 4-ethylphenol observed in a The next step is to determine whether the more precursor forms of oak lactone in oak. ‘problem wine’ submitted to the Institute method can be applied to fined wines where Several potential precursors have been was 8000 µg/L! Further analyses by our the concentration of proteins is much lower. observed. More than 20 years ago, scientists Analytical Service of more than 700 different from the whisky industry isolated, from oak samples submitted by industry, paints a very Proteolytic enzymes and/or heat as wood powder, a compound which they similar picture. It appears that wineries alternatives to bentonite termed a ‘precursor’ to oak lactone. On the would be well advised to analyse their As reported previously, work at the Institute basis of rather limited degradation studies, products for the presence of 4-ethylphenol and elsewhere has shown that juice and they assigned a structure (a methylated in order to pinpoint any microbial problems wine proteins are resistant to proteolytic gallate ester) to this compound. Despite the that might occur, and to track the efficiency attack at temperatures below 25°C. At higher limited evidence presented, this compound of corrective measures. Our experience temperatures, other groups have reported as a precursor to oak lactone has gained shows that the ‘problem’ is widespread and losses of proteins and reduced bentonite credence through literature citation. that it can occur suddenly in wineries which requirements for juices and wines treated with previously have not had such problems. proteases. Nevertheless, the use of proteases We, therefore, set out to synthesise this combined with heat treatment is not widespread methylated gallate ester in an unambiguous in industry, possibly due to the perception that fashion. Because of the possibility that the Studies on unstable wine proteins heating under any conditions is detrimental methylation on the aromatic ring might also involved in haze formation to wine quality. This appears to be unfounded have occurred as an artefact of the original because previous work at the Institute isolation, the corresponding straight gallate Staff: Dr Elizabeth Waters, Dr Leigh Francis, (Institute publications #444, #462) showed ester was also synthesised. Monomethylgallic Ken Pocock, Dr Miguel de Barros that short time (2 or 10 minutes)/high acid derivatives are relatively uncommon in Lopes, Yoji Hayasaka, Shauna Brown, temperature (~90°C) treatment had no nature, whereas gallate esters are major Professor Peter Høj clear sensory effect on wine. constituents of oak and other woods. Previous work in this project focussed on Two unstable wines were processed with two Both the gallic acid ester of cis-oak lactone characterising the proteins responsible for different enzymes on industrial scale heat and its monomethyl analogue have now wine haze and the effect of irrigation and treatment equipment at the Highett facility been successfully synthesised. The work has mechanical harvesting on the protein of Food Science Australia. The enzymes were shown that the monomethylgallate ester concentration of juice and has been added to the wines immediately before structure originally proposed for the oak communicated in many publications. The heating at 90ºC for 1 minute. The wines lactone precursor isolated and studied 20 project has now entered its final stage of were rapidly cooled to 18ºC after heating. years ago and accepted in the literature developing alternative methods to prevent thereafter is in fact in error. A French group, haze formation and of exploiting the This combined heat and enzyme treatment with whom we have established a properties of the proteins to discriminate reduced the protein levels in both wines to collaboration, are using the sample of the juices and wines varietally. about 40% of the original levels (see an straight gallate ester we have synthesised as example in Figure 5). Heat treatment alone a reference to determine whether this Use of mass spectrometry to reduced the protein levels to about 75%. The compound is an oak component and, differentiate varieties bentonite fining requirement was reduced to therefore, potential oak lactone precursor. Since subtle differences between the approximately 35% in the combined heat Understanding of the precursor pool in oak molecular weights of proteins are readily and enzyme treated wines, compared to the is an important prerequisite for optimal use detectable by electrospray mass heat alone treatment that reduced the of oak in cooperage and winemaking. spectrometry, it is theoretically possible to requirement to approximately 60%, of the identify cultivars by the protein profile of bentonite requirement for the untreated wines.

27 Team Reports

there was no protein left after treatment for 14 days. The sensory impact of this treatment is not yet known.

Our results show that for some wines, it is, in principle, possible to reduce bentonite fining requirements drastically, apparently without gross short-term sensory impacts.

A poster describing these experiments will be presented at 11AWITC to communicate these results to industry.

Haze protective mannoproteins Haze protective mannoproteins or factors (HPFs) were discovered at the Institute in 1991 and are yeast cell wall mannoproteins present in wine (Institute publications #439, 443, 460, 464, and 471). HPFs have the potential to prevent haze formation. Previous work has examined methods to extract HPFs from yeast cells or fermentation media, and two manuscripts describing this work have been published in this reporting period (Institute publications #624 and Dr Elizabeth Waters and Tracey Siebert #639). Due to low yields these appear not to be commercially viable. The project is now Importantly, and in line with previous These experiments were repeated in June 2001 focused on producing yeast strains that over Institute work, sensory evaluation of the with a Yarra Valley from the express HPFs. wines have shown that the treatments current vintage to test whether the protein resulting in reduced bentonite fining reductions are reproducible and whether the HPFs have the potential to prevent haze requirements had negligible effect on sensory impact of the treatment is negligible for formation through their ability to change the aroma and palate in the timeframes and premium wines as well as for the commercial particle size of haze. In order to understand storage period employed in this study. wines tested in the previous vintage. mechanism better a bid to augment the HPF project through collaboration within the CRC The long-term stability of the treated wines Treatment of an unstable Sauvignon Blanc for Bioproducts was mounted and has been looks promising. After six months: wine with and without enzymes at 45ºC for successful. Dr Filomena Pettolino was • All wines subsequently fined with longer time periods has also been assessed. appointed as a postdoctoral fellow to work bentonite after heat and or enzyme Unlike short term high temperature with Professor Tony Bacic (University of treatment were stable after storage at treatment, this process has been shown by Melbourne) on the physiochemical basis for typical cellar temperatures (15ºC) and others to have a significant sensory impact. HPF action. Apart from the Institute’s in-kind typical retail temperatures (25ºC). After seven hours (the first sampling time), contribution the funds to employ Dr • Hazes were not detected in any wines the combined heat (45ºC) and enzyme Pettolino are derived from CRC for after storage at 15ºC. treatment reduced the protein level to about Bioproducts and Dr Bacic’s group is of • At 25ºC hazes were only detected in 30% of the original levels. Heat treatment outstanding quality. untreated wines and wines treated with alone reduced the protein levels to about one of the enzymes. The heated wines 80%. After two days, the combined Vanessa Stockdale submitted her PhD thesis and wines treated with the other enzyme treatment had reduced the protein levels to in April 2000 and qualified for the award of have not given hazes so far even though 10% with the heat alone reducing the level the degree of Doctor of Philosophy in these wines contain unstable protein. to 70% of the original. With one enzyme November 2001. The thesis will not be

120 0.9 0.8 100 0.7 Control 80 0.6 0.5 Heat treated 60 0.4 40 0.3 Enzyme and heat Protein (mg/L) treated 0.2 20

Bentonite requirement (g/L) 0.1 0 0 2000 Chardonnay 2000 Semillon 2000 Chardonnay 2000 Semillon

Figure 4. The effect of heat and or enzyme addition on the concentration of protein and the bentonite required to stabilize a Chardonnay and Semillon wine.

28 available in the public domain until the IP selected times. After 18 months storage of example in Figure 6). The difference in regarding the identity of the HPF genes has the bottled wines at HWSL the following browning between wines with and been protected. conclusions were drawn: without ascorbic acid was small (maximum • The phenomenon of random oxidation of 0.014 AU at 420 nm after 18 months) Shauna Brown has cloned the yeast HPF was not yet evident. and was expected to be difficult to pick by genes (HPF1, HPF1’ and HPF2) and successfully • Storage position (upright versus inverted) eye. This is to be confirmed. deleted a single copy of all three in haploid had generally no impact on the level of and diploid lab yeast. Shauna is now browning in either wine type under the A much smaller and incomplete set of assessing the relative importance of each stable temperature and humidity storage is stored in an office at AWRI. The gene for the production of HPF activity. conditions used in this experiment. For temperature is controlled at 23ºC during Preliminary phenotypic analyses have one of the natural cork closures there was business hours but uncontrolled at other suggested that the HPF mannoproteins may greater browning if bottles were stored times, and there is no humidity control. have some impact on the ability of yeast to upright. The difference in browning These wines are consistently browner than tolerate certain fermentation stresses. If this between bottles stored upright or inverted the equivalent wines stored at HRWSL (for survives closer scrutiny, this is an unexpected was small (maximum of 0.006 at 420 nm example see Figure 6). Similar to the wines bonus from the HPF project and illustrates after 18 months) and likely to be in HRWSL, the difference in browning the serendipitous nature of research. undetectable by eye. In the inverted between wines with and without ascorbic storage position, this closure is showing acid was small (maximum of 0.017 AU at considerable seepage. It is possible that 420 nm after 18 months). The relative Studies on random oxidation of bottled the higher moisture levels in the cork importance of discussed variation and wines and/or in the cork/bottle interface in the absolute temperature to the enhanced inverted storage position relative to the browning of the office-stored wine, is yet to Staff: Dr Elizabeth Waters, Dr George upright position is contributing to these be elucidated. It should be noted that the Skouroumounis, Dr Mark Sefton, Dr closures in the inverted storage position brown colour in the office-stored wine Leigh Francis, Dr Zhong Kui Peng, being a more effective barrier to oxygen. would now be judged too intense by most Mariola Kwiatkowski • For both wines, and at all analysis times, individuals (Institute publication #577). there was greater browning if ascorbic Random post bottling oxidation shows itself acid had been added to the wine (see an In summary, this trial has demonstrated that in white wines as an obvious browning in a • Upright or inverted storage position had proportion of the bottled wine typically after only a small or little impact on wine six to 18 months’ storage, and is oxidation for either wine style when accompanied by loss of SO2 and ascorbate bottles are stored under ‘best practice’ as well as the development of oxidised conditions (humidity and temperature flavour. The estimated costs to the industry is control) for 18 months. $160 million per year in spoiled wine. We • Addition of ascorbic acid to both wine have worked in this area since 1995 and, styles increased the level of browning. excluding oxygenation of wine due to poor bottling procedures, have identified variable The sensory impact of ascorbic acid addition oxygen permeability of the closure as a after two years storage will be determined in major cause of the problem. It is possible August 2001. Immediately following bottling that other winemaking techniques such as and after six months storage there was no use of ascorbic acid, and upright storage, discernible difference in nose or palate contribute to the manifestation of random attributes for wines with and without oxidation (see e.g. Institute publications ascorbic acid addition. #577 and #595). This aspect and also an Figure 6. View of the top of the natural cork ‘window’ understanding of why closures are bottles. Araldite was applied to the top of the bottle to It has not been possible to determine the cover the closure bottle interface. A circle of approximately permeable have been the focus of the 30% of the original cork surface area was left exposed in impact of ascorbic acid addition, wine style project during the past 12 months. the centre of the closure. and storage position on the incidence of

The effect of ascorbic acid, bottle position and wine type on oxidation 0.220 A Riesling and a wooded Chardonnay wine, Temperature (uncontrolled) for an experiment undertaken with 0.200 22ºC during business hours assistance from Southcorp Wines to evaluate the effect of ascorbic acid, bottle position 0.180 without ascorbic, office and wine type on the extent of oxidation, with ascorbic, office were bottled in late August 1999. The wines 0.160 with ascorbic, HWSL are stored under relatively stable without ascorbic, HWSL temperature and humidity conditions in the 0.140 Hickinbotham Roseworthy Wine Science Laboratory (HRWSL, annual temperature 0.120 Temperature 15ºC range from 16.9 to 22.1ºC and humidity range from 45 to 84%). Browning 0.100 measurements on at least 30 replicate OD 420 (a.u.)) Browning (Averaged 02468101214161820 bottles for each treatment have been taken Storage time (months) throughout their storage period and will continue for another 6 months at least. Figure 5. The effect of ascorbic acid addition and storage location on the browning of a Other analyses have and will be done at Wooded Chardonnay wine sealed with a synthetic closure.

29 TSQ Mass spectrometer Photograph courtesy of The State Library South Australia Wine Cellars, Magill: 1920 Team Reports

random oxidation because random bottles with the fully covered closures for the Other than chloroanisoles, the most oxidation, as judged by browning, has not natural closures. For the synthetic closures, common contaminants identified in wine are been evident in this trial. browning in bottles with a window was simple aromatic compounds, accounting for higher than that shown by wines in bottles approximately one quarter of the problems The reader should note that the amount of with the fully covered closures (Figure 8). investigated. These are mainly associated ascorbic acid added at bottling to these Data interpretation must be made with with contamination caused by paints, resins wines was relatively low (60 ppm) and the caution due to the small sample size, used for tank lining, and some instances of concentration of SO2 relatively high (29 however, the results suggest that the relative oil contamination. Rapid and accurate ppm). This is particularly important to bear importance of distinct routes for oxygen analytical methods using solid phase in mind when comparisons to earlier permeation into wine might differ between microextraction techniques are therefore experiments are made. In these studies natural corks and synthetics. These being developed for these aromatic (e.g. Institute publication #595) the most conclusions need to be confirmed with a compounds, which include styrene, dramatic pro-oxidative effects of ascorbic larger sample set in a custom designed ethylbenzene, isomers of xylene, acid were seen at 200ppm addition levels experiment. trimethylbenzene, tetramethylbenzene and with low concentrations of SO2. diethylbenzene, as well as benzyl alcohol, benzaldehyde and isomeric monomethyl- The inherent permeability of corks Chemical analysis of industry technical and dimethylnaphthalenes. Once these to oxygen problems methods have been fully validated, the As discussed previously, a large effort has group will be able to conduct a survey of gone into developing a method that would Staff: Dr Mark Sefton, Yoji Hayasaka, Dr Alan local wines to determine the ‘background’ allow us to reliably measure permeability of Pollnitz, Gayle Baldock, Dimitra Capone level of these compounds in wine. oxygen through closures in wine bottles, in a reasonable time. A new water-soluble trap Problem solving work carried out by the of singlet oxygen has been synthesised by group is carried out in collaboration with, General activities of the Industry Kevin Pardon under the supervision of Dr and under the direction of the Industry Services team George Skouroumounis. This compound Services section. Accordingly, the results of should be appropriate to measure oxygen this work are given in the report by Industry Staff: Peter Godden, Mark Gishen, Adrian permeating through closures into model Services as are extensive studies into the Coulter, Peter Valente, Ella Robinson, wine. George Skouroumounis and Kevin adsorption of flavour active constituents by Greg Ruediger, Heather Donnell Pardon have improved the published bag in box material. synthetic method and currently have more The continuing growth of the Australian wine than one gram of the material. Due to A breakdown of industry technical problems industry is reflected in the level of demand delays in delivery of equipment, no further investigated by the group over the past 12 for the services provided by the Institute's progress has been made on assessing the years has been carried out in order to prioritise Industry Services team (Tables 3 and 4). compound in this period. targets for the development of new analytical However, following several years of rapid methods for wine taints and other growth, the level of demand has stabilised to A small subset of wines from the bottling contaminants. Nearly half of all such problems some extent over the last twelve months, albeit trial described above have been used in a are associated with chloroanisoles, and these at what could be considered as historically pilot experiment to determine whether the are now dealt with using routine analytical high levels. The Institute is convinced that the closure or the closure glass interface is the methods developed earlier by the group for provision of these services add value and a main route for oxygen permeation. Araldite, both problem-solving and research purposes. competitive edge to the industry, but also a material well known for its oxygen recognise that maintenance of a quality impermeability, was applied to the tops of bottles over the glass and the closure to either completely cover the closures of two 0.17 replicate bottles, or to partially cover the closure so that a ‘window’ remained in the 0.165 centre of the closure for four replicate bottles (Figure 7). The amount of closure surface exposed in these ‘windows’ was 0.16 between 20% and 30% of the control no coverage closures and was in the centre of the closure. These samples have been stored at 0.155 70-80% covered HRWSL for nine months. In general the fully covered highest level of browning was shown by the 0.15 uncovered controls and the lowest level by the covered samples (Figure 8). These results are further evidence that wine oxidation is at 0.145 least partially due to oxygen permeation through closures. 0.14 In addition these data suggest that natural synthetic stelvin coverage of the tops of the bottles and closures with an easily removed material Figure 7. The effect of Araldite coverage of two closure types on the browning of wooded Chardonnay wines containing with good oxygen barrier properties ascorbic acid after 18 months storage at HRWSL. Closures were covered with Araldite after nine months initial storage. Half of could reduce wine oxidation. The the bottles from each coverage and closure type were stored upright, the remainder were stored inverted. Data from both storage positions were pooled. Sample size of no coverage for natural and synthetic closures was 60 and for the Stelvin closure browning of wines in bottles with closures was 12, sample size of 70-80% covered for naturals and synthetic closures was 8, and sample size of fully covered closures for with the ‘windows’ was similar to that in the both natural and synthetic closures was 4.

31 Team Reports

service will require increased resources, in Confidentiality is an important aspect of the the previous year, while the number of order to maintain its value and responsiveness, services provided, and is strictly maintained investigations (a figure not previously and therefore relevance to the industry and at all times. Assisted by lawyers, strict Terms reported) actually declined slightly. its development of human capital. In and Conditions for the provision of problem The number of samples which were found to addition to the direct services provided to solving services have been developed during exhibit the combined and related problems winemakers on an individual basis, the the year, which reinforces the confidentiality of hazes and deposits and microbiological Industry Services team is conducting several of the work performed. This facilitates a instabilities is disappointing, and as with projects as described below, and also frank exchange of information between the previous years, the number of these wines contribute to many other Institute projects Institute and its clients, which in turn allows that have a high pH, a low concentration of described elsewhere in this report. the maximisation of the knowledge gained sulfur dioxide, and in some cases incomplete from the provision of these services. When primary or malolactic fermentations, remains a particular problem is considered to be of unacceptably high. The issues related to Technical problem solving and consulting interest to the wider industry, the results of these wine instability problems are the first investigative work are made available to have been addressed by GWRDC-funded Staff: Peter Godden, Adrian Coulter, Peter through relevant publications. The Industry project, Targeted training of wine industry Valente, Ella Robinson Services team has contributed six Technical personnel: Compilation of a technical reference Notes and six other articles to Technical manual and delivery of complementary The provision of technical support service to Review during the past year. When preparing workshops, which is discussed in more detail the Australian wine industry, primarily in the material for publication, great care is taken later in this report. In addition, the issues were form of an advisory service which disseminates to ensure that, under no circumstances is the also discussed in an article written by the a wide range of technical information, and a name of the client, or any possible team manager in December 2000: Persistent problem solving and analysis service, identifying reference, revealed. wine instability issues in the Australian represents a significant proportion of the Grapegrower & Winemaker (Institute workload of the Industry Services team. In As outlined in the previous Annual Report, publication #645), a seminar paper presented addition, the team manages a major research sensory evaluation is an important analytical in Melbourne in July 2000, and lectures trial that is examining the technical and research tool, and is commonly used in given to Oenology students at the University performance of various types of wine problem solving investigations, and commercial of Adelaide in October 2000 and May 2001. closure, and a project which is developing work conducted by the Analytical Service. a web-based technical reference manual for The quantity of sensory analysis performed The types of investigations recorded in Table the Australian wine industry, with associated has increased substantially during the last year, 3 as 'other investigative analyses' are varied, workshops which are presented in and the team manager and the Institute's and some particularly interesting and unusual conjunction with Institute Roadshow seminars. sensory chemist have continued to develop cases have been investigated this year. During the year, the first sections of this protocols for the training and assessment of an web-based manual went live, providing levy enlarged sensory evaluation panel. A summary • two separate investigations (one paying wineries with a comprehensive of the number and type of investigations conducted for an overseas company on a trouble shooting and diagnostic guide for conducted by the Industry Services team over fee-for-service basis) concluded that the wine instability problems (www.awri.com.au). the past three financial years is presented in deposits in bottled white wines consisted Table 3. The manner in which problem of a protein-phenolic complex, the The Industry Services Laboratory analyses solving investigations are reported has been phenolic material having originated from several hundred samples each year (Table 3), changed from previous years, in order to the corks and the protein from the wines. using a wide range of routine and sophisticated more accurately reflect the nature of In both cases the deposit had formed at analytical techniques. The majority of samples investigations conducted, and the workload the cork/wine interface. It should be are wine, the analysis of which is supplemented of Industry Services staff. The total number noted that it would be considered as quite by detailed sensory evaluation by a panel of of samples analysed increased by 33% from usual for phenolic compounds to be experienced tasters. Of the remaining samples analysed the majority consist of wine additives, Samples received (98/99 and 99/00) and closures, or compounds which are suspected investigations conducted (00/01)a to have caused taints and or deposits in wine. 1998/99 1999/00 2000/01 The aim of the service is to offer remedial and preventative advice based on the cumulative Identification of hazes and deposits 79 114 135 problem solving experience of the staff, and the Microbiological investigations 48 85 50 practical winemaking experience of the team Sensory assessments 52 29 51 Manager, Oenologist and other staff members, rather than providing a simple diagnosis of the Taint problems 117 186 66b cause of the problem. Increasingly, staff Other investigative analyses 245 252 128 regard their role as educational, seeking to b disseminate information in a variety of ways in Closure-related investigations 154 122 24 order to prevent the recurrence of particular Total 695 788 454c types of problem. The Industry Services athe manner in which sample numbers have been calculated for Table 3 has changed from previous reports. Some team also provides technical support to the investigations require the analysis of multiple samples, while, in some situations, one investigation is conducted on Institute's Analytical Service, particularly in several different wines from the same winery which all exhibit the same problem, and all become the subject of a the maintenance and auditing of the quality single report. Therefore, in future, the actual number of investigations conducted will be listed in the body of the management system, contract trial- Table, while the number of samples analysed will be provided as a footnote. bmuch of the decrease in the number of 'taint problems' and 'closure related investigations', many of which involve winemaking services, and the interpretation the analysis of multiple samples, can be attributed to a portion of this work now being conducted by the Institute's of analytical results. The Analytical Service Analytical Service on a commercial contractual basis. also supplies chemical analysis on problem ca total of 1048 separate wine samples were analysed as part of these investigations solving and research samples to the Industry Table 3. Summary of the number and type of problem solving investigations conducted by Industry Services team, on a contractual basis. Services during the past three years

32 extracted from corks into wine, and samples analysed being between 350 and concentration of 1.91 g/L of volatile therefore the issue lies with the need for 400 micrograms per litre, with the highest acidity (VA) expressed as acetic acid. winemakers to take this factor into concentration being approximately 600 Analysis of the wine further revealed the account when protein stabilising their micrograms per litre. These presence of numerous, viable, catalase wines. Some older winemaking texts concentrations would almost certainly be positive, rod-shaped bacteria. The cell advocate the addition of 'tannic acid' above the sensory detection threshold for morphology and catalase activity of the when conducting heat stability tests, this wine type. bacteria was consistent with the presumably to simulate the conditions in behaviour of acetic acid bacteria, but it is which wine may be placed once it comes • the number of wines that are found to be difficult to distinguish between into contact with corks after bottling. contaminated with unintentional substances gluconobacter sp. and acetobacter sp. that are not permitted additives to wine is However, since acetobacter are more • the previous Annual Report stated that of concern. During the first six moths of tolerant of ethanol than gluconobacter, three cases of oxidative pinking had been 2001, nine cases of refrigerant brine and since gluconobacter are normally investigated, two of which were in bottled contamination and six of hydraulic oil isolated from grapes and must, have a wine. In each case, the wineries contamination have been investigated. preference for sugar and appear to be concerned had not previously experienced Once again, the issues related to the intolerant of the concentration of ethanol a pinking problem in wines made from minimisation of the risks of this type of present in the wine in question, it is likely fruit from the same vineyards, as those contamination are addressed in the that the bacteria isolated from the wine that were being investigated. It was workshops described above, and at least were acetobacter sp. It is further possible ascertained that, in each case, the only two of the wineries affected by the recent that that both gluconobacter and change in winemaking procedures that cases had personnel who attended the acetobacter species were present on the had occurred was the reduction or workshops held in November and December grapes, and that gluconobacter elimination of the use of ascorbic acid. 2000. These wineries have since reported predominated early in the fermentation The incidence of this problem increased that they have now implemented the causing the high VA, and then during the remainder of 2000, with a advice given in the workshops, and that if acetobacter became dominant total of ten such cases being reported, they had taken this action sooner, then their equal to the total number of cases particular cases of contamination would • another case of high VA investigated investigated during the previous nine not have eventuated. As discussed below, during the year was also of interest, and a years. In seven of these cases the wineries a new gas chromatographic analytical Technical Note based on the investigation indicated that they had reduced or method for the identification and was published in the June 2001 issue of eliminated the use of ascorbic acid in the quantification in wine of the concentration Technical Review (issue #132). This case vintage in which the wines were made. of propylene glycol, a common coolant used involved a Shiraz wine, one barrel of 25 of While only two cases have been reported in winery refrigeration systems was developed which had developed a concentration of in the first half of 2001, previous during the year, to aid these investigations. VA above 3 g/L. The wines in ten other experience suggests that oxidative pinking barrels that were sampled all contained a is a seasonal (and perhaps vineyard • it is pleasing to note that the number of concentration of approximately 0.6 g/L. specific) problem and is more likely to be stuck fermentations reported to the The barrels in which the wine was stored reported in bottled white wines from the Institute has declined dramatically over were predominantly new, and when the 2001 vintage in the second half of the recent years. However, stuck or sluggish wine had last been racked out of barrel year. Winemakers are, therefore, fermentation is still considered an and blended, approximately four months encouraged to note when bottling their important issue in the industry, and it is earlier, the winemaker had not noted an wines, that the conclusions to be drawn interesting to note that five out of seven unusually high concentration of VA. from the Institute's previous research into regions chose this as a topic for discussion Numerous viable acetic acid bacteria, the ascorbic acid use (Institute publications at the Roadshow seminars presented bacteria most usually associated with VA #577 and #595), are not that the use of during November and December 2000. production in wine, were isolated from ascorbic acid necessarily needs to be This was in spite of the fact that similar each of the eleven barrels which were eliminated. Rather, everything else being presentations had been made at the sampled. Measurements of stave equal, the use of ascorbic acid requires a previous two seminars to be held in each thickness, the 'ovality' of the bunghole higher concentration of SO2 to be present of these regions. and the degree of ullage in each barrel in the wine, if accelerated long-term were also made, and the degree of taper browning is to be avoided. The first case of stuck fermentation of each bunghole was also noted, but not reported in 2001 was particularly measured. It was found that the problem • the previous Annual Report also described interesting. A Shiraz fermentation had barrel differed from the others inspected a Riesling wine that was found to contain virtually stopped after approximately 80 only by the ovality of the bunghole, which a concentration of 4-ethylphenol, the grams of sugar had been fermented, and was also noted to exhibit the greatest major spoilage compound formed by the must at that stage contained a taper. A small crack in the bung-stave on Brettanomyces yeast, of 2050 micrograms per litre (parts per billion). This was 1998/99 1999/00 2000/01 considered an unusually high concentration, especially in a white wine, Wineries 980 1152 1155 based on previous work conducted at the Government organisations 70 99 117 Institute (Institute publication #623). This type of spoilage is poorly documented in Other 419 413 359 white wine, and is almost exclusively associated with red wines. During the last Students 35 36 26 year the same compound has been found Total 1504 1700 1657 in a batch of white, bottle fermented , the concentration in most Table 4. Enquiries received by advisory staff during the past three years

33 Team Reports

one side of the bunghole was also noted, was obtained and reacted with calcium seven times higher than the average but the ullage in the barrel was less than carbonate to form calcium mucate. The concentration of nickel in the other that in many of the other barrels. It was, IR spectrum of the artificially induced samples. It was concluded that this may therefore, speculated that the calcium mucate precipitate was identical have been due to contamination by combination of imperfections of the to that of the crystals from the wine. fragments from the filling machine, or bunghole, and the crack in the bung- may have been due to differences in stave, had allowed a greater concentration • during the year there were two cases variety, soil and viticultural practices of oxygen to enter this barrel, and thus where winemakers were concerned about between the different vineyards from allow the acetic acid bacteria to become the possibility of metal fragments from which the grapes were sourced, even more active. Importantly, the winemaking equipment entering their though these were neighbouring imperfections in the barrel would not have wines, and subsequently dissolving and vineyards owned and managed by the been identified by casual inspection. causing irreversible contamination. same company.

• three cases of cork-boring insects were In the first case, small fragments of investigated during the year, bringing the foreign matter from a sulfitometer (a Winemaking consultation total number of cases of this problem to pressurised container from which sulfur four in the last eighteen months. In each dioxide, a commonly used preservative, Staff: Peter Godden, Adrian Coulter, Mark case the insects were found to be can be added to wine), were isolated on Gishen, Greg Ruediger infesting the corks in bottled wines that gauze and submitted to the Institute for were being stored without capsules or identification. The winemaker was The Industry Services team provides a labels, in bins, in what could be concerned that the fragments might winemaking consultancy service principally considered as standard warehousing contain heavy metals. The isolated through the Manager, Peter Godden, a conditions. In one case, a brand new material was analysed with the assistance qualified and experienced winemaker, Adrian purpose-built warehouse was involved. of staff from the Inductively Coupled Coulter, a Graduate in the Diploma in The four cases involved three different Plasma Atomic Emission Spectrometry Oenology from The University of Adelaide, insect species, which were identified with (ICPAES) unit, at the Waite Campus of the and Mark Gishen. Greg Ruediger, the Trace the assistance of CSIRO Entomology. University of Adelaide. The results Analysis Laboratory Supervisor, also joined indicated that the material contained a the Industry Services team on a 0.25 basis • an investigation was conducted for a large proportion of iron and some nickel during the year, and also holds the Graduate winery which produces sweet wines from and molybdenum, all components of in the Diploma in Oenology from The grapes infected with the mould Botrytis stainless steel. No heavy metals were University of Adelaide. Adrian Coulter cinerea, in order to ascertain if precipitates detected. Unfortunately, the ICPAES scan gained nine weeks of practical winemaking that tended to appear in the bottled does not detect chromium, another experience during the 2001 vintage, working wines over time, and which were thought important component of stainless steel. at a winery close to Adelaide. Greg to consist of calcium tartrate, could be However, the results do suggest the Ruediger has gained similar experience in related to the concentration of calcium in fragments originated from stainless steel. both the 1999 and 2000 . the wines. If this had been found to be It is possible that this could occur if water the case, then those batches of wine with were to come into contact with the inside Most queries received are technical in nature high calcium levels and thus a greater of a sulfitometer; causing the sulfur and arise predominantly from Australian propensity to form a deposit could be dioxide to react to form sulfurous acid, winemakers. However, many queries are identified, and processed in a way to which could cause corrosion of the also received from wine industry suppliers reduce the risk of deposit formation. stainless steel. and Government bodies, as well as a A 1993 Semillon wine made from relatively small number from the general botrytised Semillon grapes was analysed In the second case, metal fragments were public and secondary and tertiary students. as part of this investigation. The wine found to be flaking off nickel-coated Where appropriate, the query is answered contained a large amount of a white components of a bottle-filling machine. over the telephone, by facsimile or e-mail, and deposit that appeared to consist of Identification of the fragments was Industry Services staff supply approximately crystals, but existed in clumps rather than requested, as well as analysis of some five hundred technical papers or other pieces the more usual crystalline shape of salts wines that had recently been bottled of relevant literature to callers each year. such as calcium tartrate or potassium using the filling machine, in order to Similarly, the support facilities provided by bitartrate. After isolation, microscopy determine if similar fragments had research and library staff are important in revealed the deposit did consist of crystals contaminated any of these wines. The supplying relevant information to callers, and with a rhombic-like morphology, and the wine samples and a portion of the the analytical capacity of the Industry Services crystals left a white-coloured residue when fragments were also analysed for the Laboratory plays an important role in responding ignited in the flame of a Bunsen burner, presence of metals at the University of to many of these enquiries. In addition, indicating the presence of calcium. Adelaide ICPAES facility, and the staff are most of the investigations recorded in Table 3 However, the infra-red (IR) spectrum of thanked for their help in these result in a full written report being prepared the material did not match that of any of investigations. The fragments were found for the client. These reports contain detailed the calcium salts contained in the to contain approximately 85% nickel and technical information relating to the problem Institute’s IR spectral library. As the wine 10% phosphorus, which indicates that the being investigated and are written in a way was made from botrytised grapes, it was material was likely to be a type of nickel which seeks to explain the underlying causes suspected that the crystals might have alloy. The range of concentrations of of the problems encountered, and provide been calcium mucate. Mucic acid is a by- nickel in wine reported in the literature advice on how to prevent them re-occurring. product of botrytis mould growth and from various countries is large, and all of The reports are often accompanied by a reacts with calcium to form insoluble the wines analysed contained nickel number of technical references relating to precipitates, that deposit slowly. To concentrations within this range. the area of investigation. confirm that the crystals were indeed However, one of the samples contained a calcium mucate, a sample of mucic acid concentration of nickel approximately

34 The Institute often acts as a referral service, Formal ‘Roadshow’ visits were made to of protocols which relate to trials being increasingly for Government bodies, wine South Australian and Victorian regions in conducted on a fee for service basis for industry suppliers and wine journalists, having November and December 2000 (see various wine companies and industry suppliers. links to Australian and international wine Appendix 1). Up to six senior Institute staff research and political bodies. The vast store presented seven full-day seminars, each During the year, the vast majority of of information, both formal (in the John seminar consisting of a minimum of twelve resources allocated to this project were used Fornachon Memorial Library) and informal, presentations focussing on current areas of for the Closure Trial, a wide-ranging trial is a valuable resource to the wider industry. Institute research, or topical issues in the that is examining the technical performance wine industry. Each regional winemaker's of 14 different closure types. During the year, the Institute provided association was asked to select the assistance to the developers of public presentations to be made at their seminar, displays for the new National Wine Centre. from a list of approximately forty areas of In previous years the Institute's Director, current Institute activity, in order that each Sensory Scientist (Dr Francis) and seminar was closely tailored to the interests winemaking staff have provided advice to of the audience in each region. In addition, companies involved in this project, with a a workshop developed under GWRDC view to ensuring that the displays are project 99/1 Targeted training of industry accurate from a technical perspective. personnel: compilation of a technical reference During the current year, Greg Ruediger has manual and delivery of complementary advised on the principles and practices of workshopswas delivered by Industry Services winemaking for typical white and red wine staff in five regions on the day following the styles, and has reviewed and edited flow seminar, thus a total of 12 days of Various aspects of the charts for white and red winemaking Roadshow activities were presented. performance of the closures are being processes. Many of the displays at the The 14th Advanced Wine Assessment Course examined, including those relating to each Centre will be interactive, and the public will was held in July 2000, giving another 30 closure's physical characteristics and have the opportunity to make their own participants the opportunity to develop and extraction from the bottle, chemical analysis 'virtual wine.' To this end, multiple choice test their sensory evaluation performance. of the wine in order to examine apparent questions, along with appropriate weighting This was the second Course presented under sealing performance, and sensory analysis. for the importance of various winemaking a four-day format, which includes over 40 In addition, Dr Mark Sefton's team will techniques for the creation of a range of hours of activities over the four days. Fifteen perform chemical analysis using GC/MS, to wine styles, have been reviewed and edited. leading wine show judges, journalists and examine any effect the closures may have on In addition, digital images of typical wine winemakers, assisted in the presentation of wine aroma and flavour. microorganisms have been supplied for use the Course. The demand for the Course in the displays. continues to be extremely strong, with many As demonstrated by Table 3, problems repeat customers, despite the fact that it has associated with the use of both natural cork A summary of the enquiries received by not been promoted or advertised, which is or synthetic closures continue to represent a Peter Godden, Adrian Coulter, Mark Gishen an indication of the need for practical training large proportion of the problem solving and Greg Ruediger during 2000/2001 is of industry personnel away from their individual investigations conducted by the Industry presented in Table 4. The figures show a 2.5% work environment. As in the past, several Services team, and an increasing amount of decrease in the number of enquiries received Associate Judges for the 2000 Adelaide Wine commercial work performed by the in the previous year, following steady increases Show were selected from the most successful Analytical Service. However, until now little totalling 35% over the previous five years. recent participants in the Course, and other published information has been available Thus, the number of enquiries has apparently shows have expressed interest using Course regarding the performance of synthetic stabilised at a very high level, with the results as part of their selection criteria for closures, or little data available regarding the increased demand for this service clearly new Associate Judges. The Course continues relative importance of various parameters of reflecting the expansion of the Australian wine to attract interest from the production, closure performance, by which natural cork, industry over the same period. The proportion marketing, sales and educational arms of the screw caps, currently available synthetic of enquiries received from wineries increased industry, from all States of Australia, New closures and closures which may become to 70%, continuing a trend that has been Zealand, South Africa and the United Kingdom. available in the future, can be reliably highlighted in previous Annual Reports. compared. The broad aim of this trial is, therefore, to elucidate these factors, by The Consultation and Investigative and Evaluation of new analytical techniques examining a large number of parameters of Advisory Services are supported by vineyard and of processing aids for winemaking closure performance for a broad cross and winery visits and seminar tours to all section of currently available closure types, major wine growing regions, generally Staff:Peter Godden, Mark Gishen, Adrian on an ongoing basis for up to ten years. organised in conjunction with local Coulter, Peter Valente, Ella Robinson From this data, the relative importance of vignerons’ associations. The Institute aims to various performance parameters will be visit each major Australian viticultural region The Industry Services Laboratory maintains a determined, and this knowledge can then be through such formal visits and tours every GWRDC funded project for the improvement, applied to the development of testing second, or in some cases third year, as the development and evaluation of methods of protocols by which the future performance number of wine growing regions continues wine analysis, and the evaluation of of closures can be rapidly and reliably to increase. Routine shorter visits by key winemaking processing aids and additives. evaluated and possibly predicted. In staff are also made, as opportunities arise— The evaluations take one of two forms: the addition, the trial will also produce data on frequently in conjunction with industry relative performance of commercially parameters relating to bottling and storage, events such as capital city wine shows, and available products and the evaluation of new which may then be used to elucidate the seminars held by other industry bodies. materials marketed to the industry. Industry importance of those parameters. Services staff also provide advice to the Institute's Analytical Service on the development

35 Photograph courtesy of Orlando Wyndham

Robotic arm with needle Team Reports

Chemist and sensory evaluation expert, has been responsible for the sensory evaluation work conducted as part of the trial, and is the second named author on the published paper – in recognition of his very strong and sustained contribution to the project.

Because the testing at the 24 month point of the trial was conducted at the very end of the reporting period, there was insufficient time for a full analysis of the data. However data obtained at the 21 month time point confirm the trends of relative closure performance which are discussed below, and published in detail in the AJGWR [Vol 7, #2], which the reader is recommended to consult in order to be fully informed of all relevant trial details. Inspection of the 24 month data suggests that these trends also continued to that time point.

• Wine under the closure retained the greatest concentration of sulfur dioxide (SO2) and ascorbic acid and had the slowest rate of browning, at all testing intervals Peter Godden and Adrian Coulter (including 21 and 24 months). For other closures the trend of SO2 loss relative to Senior personnel from the Australian wine The published testing protocol (Australian the screw cap closure was apparent from an industry with extensive prior experience in Grapegrower & Winemaker [425] p 59-64, early stage of testing, and was most evident the evaluation and purchase of 1999, Institute publication #593), was in the group of synthetic closures, closures have had significant involvement in expanded at the 24 months post-bottling intermediate in the conventional corks, and the design of the trial. In addition, extensive time point to include the testing of a higher least evident in the technical cork closures. consultation with closure suppliers ensured number of replicates for measurements of For several closures upright storage tended that the closures examined were representative wine composition (SO , ascorbic acid 2 to accelerate loss of SO2 from the wine, of those being used by the wine industry, concentration and browning), and for but in many cases this effect was marginal and ensured that the bottling procedures sensory analysis. In addition, sensory analysis up to and including 18 months. However, were optimal for each closure, as far as was conducted on bottles that had been at 12 months there was greater variability practical. The wine was bottled under stored in both an upright, and inverted in the data for bottles that had been controlled conditions at an ISO 9002- orientation, and SO2, ascorbic acid and stored upright, as opposed to inverted. certified commercial contract bottling facility, browning were also measured in these same with personnel who had prior experience bottles. Testing was also conducted at 18 The loss of SO2 was in general highly with the use of many of the closures. The months post-bottling (concentration of free correlated with an increase in wine browning design of the experiment was motivated by SO ) and at 21 months (free and total SO 2 2 (OD420) and the concentration of SO2 in the need to generate data of practical and sensory evaluation on the same bottles), the wine at six months was a strong relevance, with all aspects of the protocol which was additional to the testing predictor of future browning in the wine, matching as closely as possible current foreshadowed in the protocol. This extra particularly after eighteen months. Neither industry practice, with due regard for careful testing was conducted, because it appeared the concentration of dissolved oxygen at experimental design and appropriate from previous results that the development bottling (0.6–3.1 mg/L), nor the physical replication. The wine used for the trial was of the wine had reached a crucial stage. closure measures were predictors of future produced during the 1999 vintage, from browning, and the concentration of dissolved sound Semillon grapes grown in the Clare The large amount of work that has been oxygen at bottling was not a strong Valley of South Australia, was fermented to involved in all aspects of the planning and predictor of later SO2 loss from the wine. dryness and was made utilising common implementation of this project culminated in Australian winemaking practices for wines of the submission of a large publication late in Seven of the closures displayed a strong this style. Importantly, measured variables of the reporting year. The paper, which correlation between the concentrations of composition at bottling indicate that the wine reported the results obtained up to the 20 free and total SO2 and ascorbic acid (P < had a low pH (3.1), and contained adequate month time point after bottling, was 0.001) at 12 months, but with the other SO2 (free 30 mg/L total 95 mg/L) to prevent scheduled to appear on July 12, 2001 closures the correlation was weak, or did premature oxidation. In addition, the presence published as the entire contents of the not exist at all. In addition, wine sealed of laccase, an enzyme which can cause Volume 7 issue #2) of the Australian Journal with many of the closures contained a accelerated oxidation of wine, was not detected. of Grape and Wine Research (AJGWR) higher concentration of ascorbic acid (Institute publication #666). This trial has when stored upright as opposed to The study is ongoing, and sufficient wine was been a major team effort, and Industry inverted, although there was only a bottled so that the testing of many variables Services staff acknowledges the assistance of statistically significant difference for three may continue for up to ten years. Results will the many members of Institute staff, and of the closures. These observations may be published periodically, and testing will personnel from other organisations, who imply that some closures themselves play continue for as long as it is considered that have been involved in the project. Dr Leigh a role in the degradation of ascorbic acid. useful data is being generated. Francis, the Institute's Senior Research

37 Team Reports

• The closures differed widely in regard to higher oxidised aroma. These trends of SO2 in bottled wine during storage could physical characteristics. All but one of the continued, and tended to be reinforced be used by wine companies, distributors and synthetic closures, and one of the technical for all closures, at 21 and 24 months. retailers, to identify batches of wine that are corks, were found to elongate to a greater approaching this critical level, and thus take extent than the natural corks after insertion Trichloroanisole (TCA)-type taint was a the appropriate action in the marketing of into the bottle. One of the synthetic noticeable problem for some cork and those wines. It is possible that by increasing closures was found to contract upon technical cork closures, at all testing the concentration of SO2 in wines at bottling insertion. The elongation observed raises intervals up to and including 24 months. to compensate for the losses over time possible issues related to achieving a suitable Any plastic-type taint appeared not to be identified in this study, winemakers could headspace (the space between the closures a problem with most synthetic closures up increase the longevity of their bottled wines. and the wine) when bottling wine. to 18 months, but ratings for the attribute glue-like / plastic by our highly skilled For a broad range of performance measures, In general, synthetic corks appeared least panel, were notably higher for many of variability among bottles from the same ‘consumer-friendly’ in terms of the higher the synthetic closures at 24 months. closure was not a serious issue, except for extraction forces and energies required to the TCA-affected samples, but there was remove them from the bottle, and ease of Whilst the ROTE closure was rated as some variability evident in the SO2 closure re-insertion into the bottleneck highest for the fruit attributes at all concentration, where several closures had after extraction. In addition, three of the testing intervals up to and including 24 relatively large variation. Whilst no closures synthetic closures required a torque to months, a reduced (rubbery or sulfidic) could be considered to be showing remove them from a corkscrew that was character began to be evident in wine unacceptable variation in SO2 concentration, approximately double that required for the sealed with some closures (in general indicating that so-called 'random bottle natural corks (while some synthetic closures those which had tended to retain the oxidation' was not apparent up to 18 required less torque than the natural greatest concentration of SO2) at 18 months of testing, there was greater corks). However, there was a trend for months, and was significantly higher in variability in the SO2 concentration in bottles natural cork closures to exhibit larger wine sealed with the ROTE closure stored upright. In addition, the closures that variability in these physical characteristics compared to all other closures. These did exhibit higher oxidised aroma at 18 than technical cork and synthetic closures. trends continued at the 21 and 24 month months also tended to display greater bottle- time points, with the intensity of the to-bottle variation on the rating for oxidised. All of the closures were found to expand reduced attribute increasing. in diameter by a similar amount during Closures that scored highest for the It will be of interest to note if this observation the 30 minutes following extraction, and oxidised attribute at 18 months also tended is manifested as greater variability in wine there was no relationship between degree to display the greatest bottle-to-bottle development at later time points. of post extraction expansion or the variation in the rating of this attribute. chemical and sensory properties of the Overall, technical cork closures were found wine sealed with each of the closures. Conclusions to exhibit less variability than synthetic Likewise, there was no relationship Differences in bottled wine composition that closures, which in turn tended to be less between the physical measurements of result from the use of closures of different variable than natural cork closures, when maximum extraction force, total extraction types have been assessed, and importantly, assessed for a number of the physical and energy, and the torque to remove the the study has allowed the compositional wine composition variables. However, there closure from the corkscrew, and the measures of SO2 and degree of browning to did not appear to be a trend for either chemical and sensory analysis of the wines be directly related to the sensory properties moulded or extruded synthetic closures, sealed with each of the closures, when of the wine. It is apparent that many of the when considered as groups, to be more or data for a closure which displayed notably closures included in the study are suited to less variable than each other. high extraction force, and notably poor short term (approximately 6 to 12 months) SO2 retention, (and lowest density) was storage of wines, but for longer time periods One objective of this study was to identify removed from this analysis. In addition, there is doubt over the abilities of various the most important variables of closure when data from another closure which closure to act as an adequate seal, with performance, and then apply this knowledge displayed notably high torque, extraction consequent effects on antioxidant levels, to the development of protocols that can be force and density was also removed, there browning and on oxidised aroma. It was used by the wine and closure industries for was found to be no relationship between apparent early in the trial that the closures the rapid and reliable testing of new or the density of the material from which the showing lowest SO2 concentration were existing closures. Sulfur dioxide concentration closure was manufactured, and the rated as sufficiently high in oxidised aroma was found to be a particularly important chemical or sensory analysis of the wines to consider the wine from these bottles as measurement when assessing the comparative sealed with each of the closures. markedly lower in quality. These issues performance of closures, and short-term become particularly important when it storage followed by simple SO2 or browning • Sensory analysis indicated large differences comes to managing stock on shelves. measurements provided an excellent in wine flavour properties, with closures indication of later sensory performance. which tended to result in the best The apparent importance found in this work retention of free SO2 having wine sensory of a critical concentration of free SO2 No single closure tested in this study was scores for citrus that were generally high (approximately 10 mg/L) below which wines considered entirely suitable by all criteria whilst scores for the attributes developed / are perceived as substantially affected by assessed, for the long term , oxidised were low. The situation was oxidised aroma is of considerable interest. although the ROTE was the best performing reversed for wine under closures that This observation requires confirmation to closure in terms of chemical measures of performed poorly in the retention of free determine whether this might be a generally wine composition, and sensory evaluation. It SO2. It was found that below a critical applicable cut-off value, or at the least might is possible that the reduced character which level of free SO2 remaining in the wine be appropriate as a guide for white wines of was particularly identified in wine sealed (10 mg/L) closures exhibited substantially the style used in this study. If this observation with this closure, was at least in part were confirmed, the periodic measurement exacerbated by the manner in which this

38 closure was used which, while still complying In response to the high number of cases of and Fenitrothion in grapes and marc by GC- with the closure's specifications of use, did refrigerant brine contamination reported to MS; Fludioxonil, pyrimethanil, fenhexamid, seek to exclude oxygen from the headspace. the Institute, as described above, a method tebufenozide and cyprodinil in wine, juice, for the determination of propylene glycol in grapes and marc by solid phase extraction The incidence of ‘cork taint’ (TCA wine, using gas chromatography (GC) has followed by HPLC, Eugenol in wine and contamination of the wine) was considered been developed and validated. Previously, wood products by GC-MS; Spiroxamine, an important factor with all four of the cork- the chief analysis available to Industry Services indoxacarb, trifloxystrobin, axoystrobin in based closures tested. Wine sealed with one staff when investigating cases of this sort wine, juice, grapes and marc by GC-MS; closure was affected by a styrene-like off- was the concentration of methanol, a common Trifloxystrobin, axoystrobin in wine, juice, flavour when assessed at the six month time component in refrigerant brine solutions. and grapes by GC-MS; Fludioxonil in juice point. However, due to the very poor However, this was considered unreliable in and wine by GC-MS, and, Cyprodinil in wine retention of SO2 displayed by this closure, it identifying low levels of contamination. Two and juice by liquid-liquid extraction followed was excluded from further testing. of the five cases of suspected contamination by HPLC. In addition, NATA accreditation analysed since the method was developed was obtained for the following methods: Note: Readers are asked to keep in mind were found to contain concentrations of Determination of ochratoxin A in wine by that the current study is ongoing, and that propylene glycol far in excess of the normal HPLC, and, Malathion and Fenitrothion in the results reported here generally represent levels reported in the literature. wine and juice (pending final NATA approval). the performance of the closures under This work is conducted by the Institute's examination only up to 20 months post Good progress has been made on the Quality Liaison Manager, Mr Mark Gishen, bottling. It is possible that, over time, the development of an assay for the identification who is part of the Industry Services team. relative performance of various closures may of isovaleric acid in wine. Isovaleric acid is change, or that closures which have reported in the literature as an important apparently performed well to this point may spoilage compound produced by Flavour scalping be found to perform less well in future Brettanomyces yeast, and a number of wines testing. Readers of the Annual Report submitted to the problem solving service are Staff: Dr Mark Sefton, Alan Pollnitz, should also refer to the full publication of thought to possibly contain this compound. Dimitra Capone the closure study (Institute publication #666) to understand the full context in which this Many wines are also submitted to the problem A detailed study on the capacity of two-litre study was performed. solving service that apparently contain sulphur bag-in-box packaging to absorb wine flavour compounds which cannot be removed by compounds has been completed. Changes in the addition of copper sulphate, a legal and wine composition resulted both from Analytical method development relatively common technique employed hydrolytic reactions in the wine (determined by and evaluation worldwide for the removal of hydrogen sulfide measuring changes to the wine stored in sealed and mercaptans from wine. Therefore, the glass ampoules) and from contact with the Staff: Peter Godden, Mark Gishen, Adrian development of methods for the identification packaging. Some wine components were Coulter, Peter Valente, Ella Robinson and quantification of these compounds absorbed rapidly and near-completely from would be of great benefit. During the year a wine by the plastic components of the Although the majority of resources allocated literature review has been conducted on the packaging, while others were essentially to the evaluation of methods and processing analysis of sulphur containing compounds in unaffected by this process. aids project has been spent on the closure wines, ranging from the simple volatile trial during the last year, some development compounds such as sulphides and The volatile wine components that were the of methods has also been conducted. mercaptans to more complex and less most efficiently removed were the least volatile thiol containing compounds, with a polar. Thus, 1,1,6-trimethyl-1,2- During the year, the Industry Services team view to the development of such methods. dihydronaphthalene (TDN), which has a commissioned a new Perkin Elmer Fourier kerosene-like aroma and can be an transform - infra-red (FT-IR) spectrometer, a The superiority of the new FT-IR spectrometer important contributor to the aroma and new Agilent 1100 Series High Performance compared to the previously used dispersive flavour of Riesling wines in particular, was Liquid Chromatography (HPLC) system, and IR instrument has been demonstrable. Spectra slowly generated from precursor forms in a a reconditioned Hewlett Packard 5890 Series are obtained in a matter of seconds using Semillon wine stored in the sealed glass II gas chromatograph (GC). The assistance the new FT-IR and are stored as digital ampoules for 32 days, but more than 97% of the GWRDC in providing capital funding for information. This facilitates a range of of this compound was absorbed when the this equipment is acknowledged as is the very interpretive techniques including matching same wine was stored under identical substantial contribution of Analytical Service. with reference spectra from spectral libraries, conditions in bag-in-box packaging (figure 8). identification of mixtures of dissimilar Seventy five percent of this absorption took The Institute’s internal Quality System (based materials, and quantification programs. The place following the first 24 hours of contact. on NATA requirements) required that the Industry Services team is continuing to compile Naphthalene, which is occasionally found as analytical assays previously performed using its own spectral library using the new a trace taint compound in wine and has a the old HPLC and GC systems, were validated instrument, which has been useful in structure similar to that of TDN was less for the new equipment. Validations of key identifying a number of deposits and increasing efficiently absorbed. Nevertheless, some 70% of assays required for the closure trial and the confidence of positive identification of this compound was removed by the packaging, problem-solving investigations, ascorbic acid samples, compared with the previous system. again, mostly within the first 24 hours. and the 'organic acid profile,' were the first validated for the HPLC, whilst the methods In conjunction with the Institute's Analytical A series of ethyl esters produced during for methanol, ethanol and ethyl acetate Service, the following analytical methods fermentation and including several which are were validated for the new GC system, and have been validated during the year, to the major contributors to the fruity character of validation of the GC method for standards required by the Institute's internal wine were included in the study. The acetaldehyde has commenced. Quality System, which is based on NATA concentration of these compounds changed requirements and ISO Guide 25: Malathion during storage of the Semillon wine in sealed

39 Team Reports

wine and is found in highest concentration in Pinot Noir wines, was absorbed over 32 days, and 27% of cis-rose oxide, which is associated with a lychee aroma, particularly in Traminer wines, was absorbed over the same period. A more than 50% decrease in ß-damascenone took place in both glass and bag-in-box packaging. In this case absorption phenomena were not responsible for the change in concentration.

A variety of flavour compounds associated with oak products were unaffected by storage in the bag in box packaging. These were the cis- and trans-isomers of oak lactone which, depending on concentration, give coconut, vanilla and woody characters to wines, vanillin which is the principal aroma component of natural vanilla, guaiacol and 4-methylguaiacol (smoky aromas), and 4-ethylphenol and 4- ethylguaiacol which are commonly formed by Brettanomyces yeasts during barrel maturation and which are associated with elastoplasts, medicinal and clove-like aromas. Dimitra Capone and Andres Hakkensson glass containers for 32 days, but not in a the remaining ester by the packaging This study with the bag-in-box packaging systematic way – most decreased slightly in resulted in most of this compound being clearly demonstrates the capacity of some concentration during glass storage, while the removed from the wine over 32 days. plastics to absorb flavour compounds from concentration of ethyl octanoate increased Decreases in concentration, attributable to wine in a relatively short time. Earlier slightly. For the most part, more substantial absorption of ethyl octanoate, ethyl research had also shown that natural cork- decreases in concentration resulted from hexanoate and ethyl butanoate were 55%, bark closures could similarly absorb storage in the bag-in-box packaging, and 30% and 10% respectively. chloroanisoles from wine, albeit slowly these decreases were greater for the longer (Institute publication #616). This indicated chain (and therefore less polar) esters. In the Absorption played less of a role in the that wine bottle closures, be they natural case of ethyl decanoate, a combination of a changing concentration of several other bark or synthetic, might be able to 50% reduction in concentration resulting components. Thirty-five percent of ß-ionone, modify wine flavour by absorption, and that from hydrolysis and an 85% absorption of which can give an aroma of violets to red such absorption could contribute to flavour

Differential behavior of known aroma compounds in bag-in-box

70 60 50 40 30 20 TDN (ug/L) 10 0 012481632

Time (days)

600 CHO 500

400

300 OCH3 200

Vanillin (ug/L) Vanillin OH 100

0 0 1 24 81632 Time (days)

Figure 8. Drastic differences in the behaviour of known aroma compounds is observed when stored in bag-in-box containers. Known amounts of trimethyldihydopthalene (TDN) responsible for the ‘aged Riesling/kerosene’ character, and vanillin (a known volatile compound extracted from oak) were added to a 1999 Clare Valley Semillon and their concentration followed over time as indicated. The published aroma thresholds for the two compounds are indicated with the red line.

40 Cork extraction test Photograph courtesy of Orlando Wyndham Team Reports

changes associated with bottle-ageing. A conducted by the Viticulturist and a visiting was also undertaken, as well as preparing long term study of flavour absorption from PhD student Mr Paolo Sivilotti (visiting for material to support the WFA-funded Costs bottled wine by various types of closure has seven months from Universita di Udine). To and benefits of drinking wine–A discussion therefore been undertaken and the wines date it has been confirmed that water stress and review of Collins and Lapsley by the are approaching the end of the second year in both VSP and Scott Henry vines resulted in Centre for International Economics. of storage. A report on changes during reduced leaf water potential, stomatal these first two years of storage will be made conductance, leaf area, canopy density, shoot Submissions prepared in support of the in the following Annual Report. length, pruning weight and berry weight. Australian wine industry include: Fruit exposure consequently increased and • Draft Australian Alcohol Drinking preliminary assessment indicates that skins Guidelines of the National Health and Links between viticultural and and seeds from exposed fruit had lower Medical Research Council of Australia; oenological research quantities of tannin and anthocyanin than • National Alcohol Research Agenda of the skins and seeds from less exposed fruit. Federal Department of Health and Aged Staff: Dr Sally-Jean Bell Wine was made from the fruit and the first Care; stage in the sensory analysis is scheduled to • Foetal Alcohol Syndrome Draft Scoping During 2000/2001, the Viticulturist start towards the end of November 2002. Paper of the National Expert Advisory responded to 592 enquiries, 86 more than in Committee on Alcohol; and the previous 12 months. The majority of • PRELIMINARY DRAFT RESOLUTION these enquiries related to agrochemicals. Preparation of information on wine and ECO/REGL/96/23/Stage 3 on ingredient The Viticulturist participated in several health issues labelling and PRELIMINARY DRAFT Research to Practice workshops, in different RESOLUTION OENO/ASP/99/129/Stage 3 states, and Institute Roadshows during the Staff: Creina Stockley on a health warning for the of the year (details provided in Appendix I). Nutrition and Wine Expert Group of the This project has been funded since 1990. Office International de la Vigne et du Vin Nine thousand copies of the Institute’s annual Ms Creina Stockley, a clinical pharmacologist, (OIV) (Creina Stockley is a member of the publication, Agrochemicals registered for use assumed this position in 1991 as part of her Australian OIV delegation—Nutrition and in Australian viticulture 2001/2002 were responsibilities as Health and Regulatory Health Sub-commission). produced and the information duplicated on Information Manager. A database of research the Institute’s website. The booklet was on the beneficial and detrimental health The Health and Regulatory Information distributed with the Australian Grapegrower effects of alcohol and in particular, wine, Manager has been instrumental in organising & Winemaker, Technical Review and in the was established on the internal computer and developing the program for the wine Research to Practice™: IPM and Spray network of the Institute and as part of the and health symposium session of the 26th Application manuals. The tables are featured John Fornachon Memorial Library. This was World Congress and 81st General Assembly in Australian Viticulture. The 2001/2002 facilitated by the subscription to relevant of the Office International de la Vigne et du MRLs for Australian export markets was also medical and scientific journals, and by the Vin to be held 11–18 October 2001. The updated for the Institute’s website. formal and informal exchange of information theme of this session is "Current wine and between complementary organisations, both health information: the issues in perspective". As a direct result of the Institute’s national and international. During There are also three sub-themes: 1) The agrochemical publication and its Residue 2000/2001, 74 independent information current and future status of wine and health Analysis Service, the Viticulturist and Greg requests were received on wine and health information; 2) The need for consumers to Ruediger have been devoting an increasing issues from industry, government and the relate this information to their diet and amount of time in liaising with major chemical general public by the Health and Regulatory lifestyle; and 3) Responsible communication companies. Companies are increasingly aware Information Manager. of wine and health information. of the importance the wine industry places on meeting export market specifications. Thus, Subscription to relevant medical and other Institute and industry consumer health and they are keen to work more closely with the journals has continued. The journals have safety information and support system industry and the Institute to ensure that their been regularly scanned, the database of The Health and Regulatory Information Manager products can be used in viticultural pest and research on the health effects of wine has has also prepared the Institute’s and industry’s disease control programs in such a way that been added to and articles have been consumer health and safety information and the maximum residue levels set by Australian prepared for inclusion in the Institute’s support system, which is accessible on the export markets will not be exceeded. publication, Technical Review and for other Institute’s web-site www.awri.com.au. Australian wine industry and international With short notice, the Viticulturist coordinated alcohol industry newsletters. Articles and Project coordination and prepared the application for National other material have also been prepared for Through Creina Stockley, the Institute has Registration Authority (NRA) approved the electronic and print media, for example, played a coordinating and a participating emergency use permits for four insecticides six articles have been prepared for the role in three GWRDC-funded research on behalf of the wine industry for the control bimonthly international publication, AIM— projects on medical aspects of wine of locusts, and these permits are held by the Alcohol in moderation, and four articles for consumption entitled AWR97/2 Potential Winegrape Growers’ Council of Australia Inc. the quarterly newsletter of the Australian cardio- and cancer-protective effects and Society of Wine Education. The Health and mechanisms of wine. GWRDC pays the The investigation into the effect of plant water Regulatory Information Manager is also a Heart Research Institute, The University of status and canopy management on the member of the editorial board of AIM— Western Australia and CSIRO Health Sciences phenolic profiles of grapes and wine in Alcohol in moderation. A national radio and Nutrition directly. The respective project collaboration with Dr Patrick Iland and Dr Peter interview and a television interview were also titles are: Potential cardioprotective activities Dry (University of Adelaide), Orlando undertaken related to the issue of potential of wine components based on synergistic Wyndham (Langhorne Creek) and Professor blood products in winemaking. A review of interaction with vitamin E; Grape antioxidant Enrico Peterlunger (Universita di Udine, Italy), the draft Training Procedures Manual for the phenolics: absorption and inhibition of lipid commenced in October 2001. This trial was National Wine Education and Training Centre peroxidation in humans; and Reduction of

42 damage to LDL and DNA from oxidative free Significant efforts, energies and resources have and to the researchers of the Institute. The radicals by the regular and moderate also been directed towards ensuring the Library is also used extensively by other consumption of wine. All three projects have establishment of realistic and reasonable groups such as students, government bodies completed their third year of funding. maximum limits for the additives and processing and private companies (see Table 5). aids approved for use in winemaking by the Codex Alimentarius Commission, on behalf Information and document delivery services Technical and regulatory support to the of the AWBC, WFA and AFFA (Wine, Vine The Library has excellent access to international Australian wine industry and Citrus Section). Technical information databases, particularly in the fields of science, and/or issues that have been reviewed, and technology and medicine. If requested, the Staff: Professor Peter Høj, Creina Stockley discussion or position papers prepared or Librarian, Catherine Daniel, will carry out and Rae Blair contributed to during 2000/2001 include: the online searches on commercial databases on national system of gene technology regulation any appropriate topic (on a fee-for-service Information requests of the Therapeutics Goods Administration basis). Alternatively, Library staff can provide, One of the activities of the Institute has been and Australia New Zealand Food Authority; free of charge, a report of relevant articles to provide legal/regulatory and technical ochratoxin A; silver nitrate; tannin and indexed on the Library’s in-house databases. advice and assistance to the Australian wine tannin-like additives; current status of Requests for information from Library staff industry, through the Health and Regulatory country of origin labelling; acidification with continue to rise significantly with a 108% Information Manager, the Industry Services lactic, malic and tartaric acids; wine as a increase in information requests for team and the Director. During 2000/2001, source of dietary sodium; BSE and the 2000/2001 over 1999/2000. Most of the 110 independent information requests on Australian wine industry; and factors Library's services showed increases in technical and regulatory issues from the influencing the sodium and potassium demand, with the exception of online search government and industry were fielded by the content of Australian wines. requests and requests for articles from Health and Regulatory Information Manager. Technical Review (-3.8%). This minor Technical Information is also disseminated to decrease in requests for articles from Industry committee membership the Australian wine industry, including all Technical Review has been more than Additional support to the industry is derived grape and levy payers, by the institute’s compensated for by a 40.5% increase in from the Director’s membership on the AWBC bimonthly publication, Technical Review, of other articles forwarded over 1999/2000 Compliance and Technical Advisory Committee, which the Health and Regulatory Information figures. Database searches undertaken on International Trade and Technical Advisory Manager is editor, and by the Institute’s internet and CD-ROM networked databases, Committee and the WFA Technical Committee. Annual Report edited by the Director and now recorded as Information requests, offset One of the important aspects of the Institute’s Communication and Publicity Manager. the continuing decline in online database support of the Australian wine industry is its Australian Wine Industry searches. These specialist databases are now pivotal role in facilitating the triennial Technical Conference only used to access information not available Australian Wine Industry Technical Conference Several Institute staff members, namely, in the public domain and to deliver specialist (AWITC), in conjunction with the Australian Professor Peter Høj, Peter Godden and Rae current awareness services. The funding Society of Viticulture and Oenology. The Blair, serve on the Conference Planning approved for the third Library staff member Director is the Chair, and the Personal Committee of the Eleventh Australian Wine (whilst primarily to assist with the loading of Assistant to the Director is the Conference Industry Technical Conference, and Professor records for the web-accessible database) has Manager and Treasurer. Professor Høj and Høj and Mr Godden served on the Program facilitated an appropriate level of service and Ms Blair are also members of the Australian Sub-Committee. The formal program has response time to the steady increase in Organising Committee of the 26th World been developed and will cover eight two requests received by the Library. Congress and 81st General Assembly of the hour sessions from Sunday, 7 October to Office International de la Vigne et du Vin to Wednesday, 10 October. Some 74 Document delivery be held 11-18 October 2001. workshops are being planned, and the The Library can supply either books or workshop program, being convened by Mr photocopies from its collection or obtain At present, the Health and Regulatory Godden, will run from Sunday, 7 October to such items for wine industry clients through Information Manager is a member of the Thursday, 11 October. Institute staff will the interlibrary system. Patents or standards following committees: the AWBC Legislative conduct several workshops and prepare can also be ordered. Electronic ordering and Review Committee; the WFA Technical more than 30 posters. Jeremy Hack acts as delivery services mean that most interlibrary Committee; and the National Reference the poster coordinator. requests are fulfilled within five days. Committee-Environment Strategy Development Charges apply for the supply of some items. (SAWBIA). Ms Stockley was also elected This project of technical and regulatory support Vice-President of the Nutrition and Wine to the Australian wine industry is ongoing, Specialised information services Expert Group in March 2001. as technical and regulatory issues are regularly The Library staff continue to be actively raised by the government or by industry, both involved in the production of specialised Reviews and publications in Australia and internationally. Furthermore, information products for the benefit of the During 2000/2001, supplementary to the these issues often span several years. wine industry, such as the annual and web- Standard 2.7.4 for wine and wine product of based editions of the Agrochemicals the [joint] Australia New Zealand Food registered for use in Australian viticulture, Standards Code that was gazetted in Provision of technical information the bimonthly Technical Review (where the December 2000, a draft industry standard Librarian is Acting Editor for editions #133 for wine, sparkling wine, , wine Staff: Rae Blair, Catherine Daniel, Ingrid Oats and #134), and several in-house technical products, reduced and low alcohol wines information databases. was prepared by the Health and Regulatory The John Fornachon Memorial Library holds Information Manager in conjunction with the largest collection of wine technical Library collection the AWBC and WFA, for potential inclusion literature in Australia. The Library’s principal A total of 79 monographs and seven in the Australian Wine and Brandy responsibility is to provide technical conference proceedings were added to the Corporation Act 1980 and Regulations. information to the Australian wine industry library collection in 2000/2001. The Library

43 Team Reports

subscribes to 54 journals and receives Wine Staff Other Total % inc/(dec) approximately 70 annual reports, journals industry over 1999/2000 and newsletters through exchange and donation. The Library also maintains a Information requests 515 484 1155 2154 108% collection of over 19,000 reprints. Online database searches 5 14 19 (47.2%) Interlibrary loans Library databases • requests sent1 159 496 655 2.8% Apart from a computer-based catalogue of • requests received2 159 159 117% books and journal holdings, the Library has Technical Review requests3 167 (21%) several specialist in-house databases, which Technical Review articles forwarded4 907 (3.8%) index over 38,700 scientific and technical reprint Articles forwarded5 662 40.5% articles; over 2,000 articles on the medical Number of Institute publications aspects of alcohol consumption; and the forwarded 440 18% bibliographic details of the Library’s collection Articles photocopied 3795 n/a of the European Union wine legislation.

1 Staff at the JFML sent a request to another library for an article. The Librarian, Catherine Daniel, provides 2 Requests received by the JFML from other libraries for articles from our collection. reports, either on particular subjects or 3 Number of requests received for articles published in the Technical Review. authors, listing the records retrieved from 4 Number of articles forwarded (usually more than one article is requested). any of the Library’s in-house databases. A 5 Number of articles forwarded, excluding staff publications (usually more than one article is requested). summary of the size of the Library’s catalogue and information databases is Table 5. Summary of information requests during 2000/2001 given in Table 6.

The Library provides access to its databases, All levy payers receive a printed copy of the Library catalogue databases 24 hours a day, via the internet, to Wine and revised edition automatically, and the web- BOOKFILE: Books, Grape Levy payers only. The restriction in based edition (accessible on www.awri.com.au) conferences and theses 3,304 access is enforced to comply with copyright is updated on a regular basis. ARTICLES: scientific papers 38,737 approvals obtained from the various publishers. MEDIC: medical papers 2,304 Edited database records continue to be Technical Review posted onto the Library's web database, Technical Review is received by all Wine Levy JOURNALS: journals, newsletters, with the total number of searchable records paying wineries in Australia and, through statistics and annual reports 428 available in excess of 10,000. subscription, by government and other organisations and individuals, both in Australia Library information databases Agrochemicals Grid and overseas. Technical Review provides REGS: European Community As reported elsewhere in this Annual Report, progress reports to the industry on the wine legislation 372 Dr Sally Bell and Catherine Daniel prepared Institute’s research as well as updates on relevant the twelfth edition of the Agrochemicals conferences, regulatory amendments and ISYS – full text retrieval registered for use in Australian viticulture. medical issues. Technical Review’s ‘Current database covering United States of America Federal Register 689

Web accessible database 10,309

Table 6. Number of records on the Library’s catalogue, information and web-accessible databases

Literature’ section provides citation details and abstracts of recently published technical and scientific articles. Recipients of Technical Review may order articles featured in the ‘Current Literature’ section via a request form available within each issue. The Librarian, Catherine Daniel, was Acting Editor of Technical Review’s June issue, #133. Dr Barbara Hardy AO continues to support the publication of Technical Review through financial contribution to the Thomas Walter Hardy Memorial Trust.

Email service The Email advice and information on technical issues service continues to be a fast and cost-efficient way of disseminating important technical information to interested members of the Australian wine industry. There are over 530 email addresses recorded to receive the email bulletins to date. Twelve email bulletins were issued during the year Rae Blair, Catherine Daniel and Heather Donnell and are shown in Table 7.

44 Targeted training of wine industry based on the collective information generated 18/7/00 Launch of new personnel: compilation of a technical by various teams within the Institute. The Agrochemical booklet reference manual and delivery of workshops also contain a large amount of 18/7/00 Use of Shirlan for control complementary workshops. practical winemaking advice regarding not of Phomopsis only the causes of problems, but also their Staff: Peter Godden, Mark Gishen, Adrian avoidance, and remedial advice on how to 19/9/00 Notice of First National Coulter, Peter Valente, Ella Robinson deal with them if they occur. Environment Conference The primary aim of this project is to produce During the year, work commenced on new 3/10/00 Switch Fungicide registered for flexible and updateable information sections of the web site, particularly those control of Botrytis packages on selected technical subjects, which will deal with wine microbiology in 4/10/00 Chemical Control of Australian which will be delivered to winegrape levy more detail, with particular emphasis on the Plague Locusts in Viticulture payers via the world-wide-web, and identification of both wanted and unwanted workshops to be held in all major grape wine microflora, and good laboratory and 6/11/00 Fenitrothion emergency growing areas of Australia in conjunction winery practice with regard to wine use permit with the existing Roadshow seminars. microbiology issues. It is envisaged that this 6/11/00 Flint fungicide section will be expanded to encompass the The Institute has a considerable amount of issue of stuck fermentation, thus addressing 6/2/01 AWRI denies claims of collective knowledge pertaining to grape another cause of potential wine instability, as endorsement of NuKorc and wine production, much of it generated discussed elsewhere in this report. A further over the years by research and Industry section on which work has commenced is 16/2/01 Blood products for sale from Services' projects that have been supported the design of a winery laboratory, an area Yeruva SA, Argentina by industry research-levy funding. Although that has obvious synergies with those 2/5/01 Non thermal food processing this research has produced many technical already addressed, and the subject of an seminar advice publications, there is also a great deal of increasing number of advice calls to the information generated and recorded in a Industry Services team. 10/5/01 Eleventh Australian Wine Industry more informal manner by the staff concerned. Technical Conference – updates This project, therefore, seeks to make a The information on the web site and early registration closing record of this information, so that it can be (www.awri.com.au) is password-protected, for date notification delivered to the industry in a manner in which it use only by Australian grape and wine research 16/5/01 Eleventh Australian Wine Industry is both readily useable, and relevant to those levy payers. The password has been supplied to Technical Conference – advice of involved in day-to-day wine production. It is all levy payers via personal letter. Levy payers new workshop added to program envisaged that the project will be ongoing, and who are unsure of the password may obtain that many areas of interest to winery technical it by contacting either the Institute's Librarian Table 7. Email bulletins sent during 2000/2001 personnel will be addressed in due course. or Communications and Publicity Manager. During the year, the first two major objectives The posting of the technical information of this project were achieved; the posting of generated by this project onto a web-site The John Fornachon Memorial Library technical information and associated detailed was a large undertaking, and required a Endowment Fund problem solving guide onto the Institute's substantial amount of design and The Institute acts as the Trustee of this fund, web-site, and the delivery of the first development of the website in order to make which was established in 1969 by donations workshops developed under this project, in it both as practical and relevant as possible, from winemakers and friends of the late five wine production areas in South Australia while at the same time aesthetically pleasing. John Fornachon, the first Director of Research and Victoria. Both the web-based information It was, therefore, decided that concurrent of the Institute. The Library is funded by an and the workshops relate to wine instability development of the Institute's overall site annual grant from the Grape and Wine issues. As discussed elsewhere in this report would generate substantial efficiencies, Research and Development Corporation, the combined and related area of hazes and rather than addressing this as a separate together with the income generated from deposits and microbiological instabilities issue at a later date. Industry Services staff, investment of the Endowment Fund. continue to represent a significant proportion therefore, had a high level of involvement in of the problem wine sample investigations the development of a new web site for the Acknowledgments conducted by the Industry Services team. In Institute, in conjunction with the Institute's The Institute wishes to thank all individuals many cases it is apparent that the problems Communications and Publicity Manager, and companies who contribute to the could have been avoided if winemakers were Director, and two external design and web- collection through donations or exchange more aware of the causes of many types of site development companies. The template agreements. The support of the following instability, and were able to access practical design developed for material generated persons and organisations that have donated and relevant tools with which to trouble- under this project has been successfully books or journals is acknowledged: shoot the problems, before significant wine applied to many other areas of the site. Australian Bureau of Agriculture and spoilage had occurred. The web site and Research Economics, Australian Dried Fruits workshops have addressed these issues, Corporation, Australian Wine and Brandy providing practical trouble shooting information Waite Campus Mass Spectrometry Facility Corporation, Australian Wine and Brandy and simple diagnostic tests, with which to Producers’ Association, Commonwealth isolate and identify a wide range of hazes and Staff: Yoji Hayasaka, Gayle Baldock Scientific and Industrial Research deposits in wine. The text is supported by a Organisation, K.F. Pocock, Dr B.C. Rankine, gallery of images of deposits in wine that have The four important roles of the Waite Viticultural Publishing Inc., Winemakers’ been generated from routine problem solving Campus Mass Spectrometry Facility are to Federation of Australia Incorporated, investigations. In addition the web site act 1) as a leader in the application of mass Yalumba Winery. provides extensive background information spectrometry to grape and wine research; 2) on the causes of a wide range of instabilities, as an investigator to solve the problems written by a contracted technical writer and facing the wine industry and individual

45 Team Reports

winemakers, using mass spectrometric Research activities GC- LC- techniques; 3) as a collaborator with The Involvement in the research activity of the MS/MS MS/MS University of Adelaide in the research and Institute is one of the most important roles teaching activities involving mass of the Facility and its demand has been Staff/students spectrometry; and 4) as a provider of increasing in not only classic applications like of AWRI 90% 82% versatile and advanced mass spectrometric volatile flavour analysis but also techniques and expertise to the scientific characterisation of macro, involatile and Adelaide University 10% 15% community including public as well as labile compounds like proteins, tannins and Other Institutes 0% 3% private institutions. Much of our work is, glycosidic conjugates. The following research therefore, conducted on a fee-for-service basis. activities highlight this function. Table 8. Time usage of the GC-MS/MS and LC-MS/MS for 2000/2001 The Facility’s instruments • The use of mass spectrometry to The Facility accommodates four instruments differentiate varieties of juice was Characterization of highly polymerised seed namely a ThermoQuest TSQ gas investigated under project AWR 8 ‘Studies tannin by electrospray mass spectrometry chromatography tandem mass spectrometer on unstable wine proteins involved in haze was conducted under the projects of AWR (GC-MS/MS) installed in February 2000, a PE formation.’ Varietal differentiation of juice 96/1 and CRCV 1.2 in collaboration with Dr Sciex ionspray tandem mass spectrometer was successfully achieved based on the Stephane Vidal. The characterization of (LC-MS/MS) installed in October 1995 and composition of PR-proteins determined by highly polymerised tannin (proanthocyanidins) two Agilant gas chromatographs with mass electrospray mass spectrometry. This work is a challenging task and has been attempted selective detectors (GC-MSD) installed in was published in the Journal of using chromatographic and analytical March 1997 and January 2001, respectively. Agricultural and Food Chemistry (Institute techniques including mass spectrometry. To The GC-MSDs are used mainly for grape and publication #663) and an oral date, the sizes of proanthocyanidins wine flavour projects under the supervision presentation was made by Yoji Hayasaka determined by mass spectrometry were of Dr Alan Pollnitz, and one has been fitted at the 18th Conference for the Australia pentamer in grape seeds by liquid secondary recently with a Gerstel MPS2 autosampler and New Zealand Society for Mass ion mass spectrometry (Vivas 1996), which also has solid phase micro extraction Spectrometry (4-9 February 2001). The heptadecamer in apple (Guyot, 1997) and and headspace capability. The GC-MS/MS application of this technique for wine is docosamer in litchi (Roux, 1998) by and LC-MS/MS are used for various purposes under investigation. electrospray mass spectromery and undecamer and appropriate financial arrangements for in grape seed (Kruger, 2000) by matrix assisted all users are in place to recover the running • Search for wine pigments in red wine laser desorption ionisation mass spectrometry. cost of the Facility. Table 8 details the time using nanoelectrospray tandem mass In our studies, grape seed tannins were usage of the GC-MS/MS and LC-MS/MS. spectrometry was conducted as part of fractionated into two fractions by column project AWR 96/1 ‘Wine grape tannin and chromatography according to their molecular Problem solving work colour specification,’ in collaboration with sizes. The two fractions, LDPS (lower degree During the reporting period, thirteen cases Dr Robert Asenstorfer of the University of of polymerisation) and HDPS (higher degree of chemical analysis of either juice or wine Adelaide. Wine pigments such as a of polymerisation), were analysed by were conducted using GC-MS, in collaboration pyruvic acid adduct (vitisin A), a 4- electrospray mass spectrometry in the negative with staff of the industry services. vinylphenol adduct (pigment-A) and a mode. The LDPS and HDPS fractions were vinylcatechin adduct of malvidin-3- well characterised and the presence of more As mentioned in the previous annual report, monoglucoside have been reported in the than pentacosamer of procyanidin gallates in the development of a taint screening method past five years. All wine pigments are the HDPS fraction was demonstrated. These has commenced using GC-MS. This formed during and after vinification. They analytical techniques are sophisticated but development is conducted for the projects are substituted at the C-4 position of much needed to understand better the of AWR 19 Chemical analysis of industry malvidin-3-glucoside and thought to be tannin properties of wine. A manuscript technical problems and AWR 10 Technical more chemically stable than malvidin-3- outlining this study is in progress. problem solving and consulting. In order to glucoside. Nanoelectrospray was used for investigate the common contaminants the screening of wine pigments The considerable contribution of the Facility frequently encountered in the wine industry, (anthocyanin derivatives) in red wine. The to the CRCV tannin projects (Program 1.2) the problem solving work investigated by advantages of this technique over was made in collaboration with staff of The the Institute using GC-MS in the past 12 conventional electrospray are the better University of Adelaide. The following years was reviewed. sensitivity, the smaller size of a sample outcomes were achieved. required (1 µL) and that sufficient time for - Asenstorfer, R.E.; Hayasaka, Y.; Iland, P.G.; The review revealed that toluene, xylene, the application of various tandem mass Lambert, S.G.; Jones, G.P. Wine phenolics: styrene, alkyl (C3-C4) benzene, spectrometric techniques – is available due the development of pigments in red wine. naphthalene and alkyl (C1 and C2) to very low flow-rates. Steans, G., ed. Proceedings of the 1999 naphthalene were common as conference of the New Zealand Society for contaminants in the rare faulty juices and The screening method for wine pigments Viticulture and Oenology; 4-5 November wines submitted. These compounds are was successfully developed using 1999; Auckland, New Zealand Society for not normal constituents of grape juice nanoelectrospray tandem mass spectrometry. Viticulture and Oenology; 1999: 83-78. and wine but they are volatile This method was applied to search for new - Asenstorfer, R.E.; Hayasaka, Y.; Jones, G.P. components of some petroleum possible wine pigments in red wine extracts. Isolation and structures of oligomeric wine products. Therefore, these compounds Apart from the wine pigments reported pigments by bisulphite-mediate ion were nominated as target compounds of above, the possible presence of malvidin-3- exchange chromatography. J. Agric Food the taint screening of juice and wine. The monoglucoside hydroxy, vinylalcohol, 4- Chem. Submitted. method development is in progress. vinylcatechol and vinylsyringol adducts were - Kennedy, J.A; Hayasaka, Y.; Vidal, S.; Waters, demonstrated. A manuscript outlining this E.J.; Jones, P.J. Composition of grape skin study is well progressed. proanthocyanidins at different stages of berry development. Manuscript in preparation.

46 Grape contest: 1973 Photograph courtesy of The State Library of South Australia

HPLC instumentation Team Reports

of assisting control and availability of policy and procedural documentation, as part of a general move toward quality management systems implementation.

As reported elsewhere, Mark Gishen remains heavily involved in the collaborative research project evaluating the use of near infrared spectroscopy (NIRS) for the rapid determination of a number of compositional parameters in grapes, must, wine and grape spirit, and recently assumed primary responsibility for the project. The details of this project are reported elsewhere in the Annual Report.

Publications and seminars given by the Quality Liaison Manager and staff working on the NIR project are shown in Appendix 1.

Communications and publicity

Staff: Rae Blair

The Communication and Publicity Manager’s role is a part-time position for Rae Blair, who

Gayle Baldock and Yoji Hayasaka is also the Personal Assistant to the Director and Conference Manager of the Eleventh Quality Liaison Manager Institute's new web site to enhance its Australian Wine Industry Technical Conference. circulation throughout the industry. As Communication and Publicity Manager, Staff: Mark Gishen Mark Gishen is assisting in the staging of she is responsible for ensuring that industry events of interest to the wine industry and stakeholder groups clearly comprehend The major output of the Institute’s activities in including workshops on objective grape the positioning (value) of the Institute. This the provision of advice on quality management quality measurement, quality management positioning is developed in line with the techniques to industry remains the From options, HACCP plan development, and NIR Institute’s mission statement and business Grapes to Glass program jointly run with the spectroscopy calibration for the 11th plan objectives. Part of her role is to South Australian Wine and Brandy Industry Australian Wine Industry Technical coordinate the Institute’s printed material Association. The program was published in Conference (Adelaide, October 2001), and is and other non-technical communications, August 1997 and enhanced with a simple contributing as part of the speaker program and to act as a conduit for media. The HACCP module in 1999. Industry interest for the same conference. Communication and Publicity Manager is remains greatest in the HACCP module — a also responsible for the performance and simple program delivered in a one-day course Mark Gishen takes primary responsibility for output of the John Fornachon Memorial that incorporates an HACCP-type (hazard the internal quality management systems of Library. The report of the activities of the analysis and critical control point) food safety the Analytical Service, overseeing management Library can be found elsewhere in this Report. plan. This module was designed to satisfy reviews, documentation, auditing, and the requirements of the proposed (now corrective actions. An external reassessment of Technical Review delayed) changes to the food hygiene the laboraotry's quality system was conducted The Communication and Publicity Manager regulations, and meet the needs of the by NATA during this year. The method for the has overall fiscal responsibility for the smaller scale businesses in the industry. determination of ochratoxin A was successful production and supply of Technical Review, Three courses for the HACCP module were in gaining accreditation, and several changes and the pricing structure of Technical Review conducted throughout the year, adding 20 to signatory status of staff were approved. It was assessed in line with the introduction of more to the growing list of companies was pleasing to note that improvements to GST as at 1 July 2000. During the reporting having attended (now totalling 73). The the system that have been implemented period the cost of subscribing to Technical From Grapes to Glass program provides a recently meant that the system already complies Review was further considered and no price simple and relatively cheap program that with many of the elements of the revised rise was deemed necessary, although the uses a staged approach in the attainment of standard, ISO 17025, ahead of the cost structure of recouping postage costs internationally recognised standards, starting requirements of NATA. The Analytical Service was streamlined. The subscription period from the Codex HACCP principles and continues to participate and excel in both was also set at expiring on 30 June each year. leading to the full ISO 9000 quality national and international proficiency testing The Communication and Publicity Manager management standard. The program may programs for routine wine analysis and for met with the Director, Health and Regulatory soon be upgraded and simplified to reflect agrochemical residue testing. The Analytical Information Manager (as Editor of Technical the changes brought in with the new year- Service continues to manage its quality Review) and the Librarian to discuss 2000 version of the ISO 9000 standard. management system with the aid of the changing the format of part of the Technical Paradigm Quality software package and will be Review to ensure its copyright-compliance. The Code of good manufacturing practice upgrading to a newer version release of the that was developed during the development software in the coming year. Following the Coordination of media of the HACCP module of the From Grapes upgrade, the software will be introduced Institute staff were interviewed by to Glass program, was also published on the throughout the Institute, primarily as a means representatives from various forms of media

48 over the year as shown in Table 9. Date Staff Discussed Media On Monday and Tuesday, 5 and 6 February, the Communication and Publicity Manager 3/8/00 PWG Brettanomyces Tim White/Australian circulated a press release (prepared by the Financial Review Manager Industry Services and the 13/10/00 PWG Addition of tannin to wine Dan Sogg/Wine Spectator Communication and Publicity Manager) to correct some information relevant to Several media interviews were conducted by Peter Høj and Mark Gishen as a result of a NuKorc, to 62 wine industry organisations stand-alone press release issued by BRL Hardy regarding the Near Infrared Spectroscopy project and wine media Australia-wide, and to 358 14/10/00 PBH Research in the Australian Wine Industry Sky TV (UK) email addresses via the Institute's email 8/8/00 MG Grape quality/NIR ABC Regional SA bulletin. The release was also published in (Country Hour) the February edition of Technical Review. 9/8/00 MG Grape quality/NIR ABC Tropical North (Drive) Editing 11/10/00 SJB Chemical control of locusts ABC 5CK the North The Communication and Publicity Manager & West Report assisted the Analytical Service staff by 27/10/00 ADC ‘Sour sob’ character in Sauvignon Blanc wine Philip White/ proofing their 2000-2001 Analytical Fee The Advertiser Schedule and with the Director she also 6/11/00 ILF AWRI Stefan Maus/ compiled and edited the 2000 Annual Wein-Wissenschaft Report. The Communication and Publicity Manager coordinated the production of 8/11/00 ILF Flavour chemistry Paul Clancy/Australian Agrochemicals registered for use in Viticulture and Australian Australian viticulture 2001/2002. Industry Journal Institute’s website 10/11/00 PBH Silver nitrate in wine Jeni Port/The Age Much of the year was spent on the 12/12/00 PBH Pesticide spray and residues Philip Satchel/ABC 5AN evaluation and implementation of the design of the new Institute website and preparation 22/2/01 CSS Blood products in wine Natalie Larkin/ of information to be loaded onto the ABC Radio National website. It was decided that the Institute 22/2/01 CSS Blood products in wine ABC TV News needed a separate domain from the 18/1/01 PWG Closure trial and AWRI Mark Stean/The Advertiser Adelaide University’s server, and accordingly, 27/1/01 SJB Conventional versus organic viticulture Nick Galvin/ the Communication and Publicity Manager ABC’s Organic Gardener arranged for the Institute to purchase server space from Internode, and has registered its 1/2/01 PBH Impact odorants in wine Max Allen/The Australian own domain name awri.com.au 15/2/01 PWG Closure trial Eric Cummins/ National Grapegrower Accordingly, the address for the new website 2/3/01 PWG Closure trial Laura Lee Madonna/ is www.awri.com.au www.WorldWineTrade.com 9/3/01 ILF Mouthfeel wheel Mark Steene/The Advertiser The sitemap was designed using four simple, 12/3/01 PBH/MGNear Infrared Spectroscopy project Natasha Mitchell/ABC user-friendly boxes: ‘Research,’ ‘Services,’ 16/3/01 ILF Mouthfeel wheel Cindie Lange/ ‘About Us’ and ‘Information.’ Each box National GrapeGrowers contains various sub-sections and can be 20/3/01 PBH Genetically modified organisms in vines/yeasts. Florence Kennell/La Revue accessed by rolling the cursor over the box du Vine de France and to reveal a slide-out menu of sub-sections. www.magnumvinum.fr The Director had strong input into the final 16/4/01 PBH AWRI Channel 9 Directions product, along with the Communication and 16/4/01 PWG Closure trial Channel 9 Directions Publicity Manager, the Manager – Industry 31/5/01 PBH Chemicals used in vineyards Ken Dickin 5DN Services and his staff. The site was designed by Geoffrey Reed Communications, and was 31/5/01 PWG Closure trial Rose Murray Brown facilitated by WineBiz. Table 9. Staff interviews by media 2000/2001 Website maintenance Various staff within the Institute are charged Eleventh Australian Wine Industry who is responsible for the registration with ensuring the website is kept up to date. Technical Conference process, and who had processed a record The ‘Research’ section is maintained by Leigh Full details of the Eleventh Australian Wine number of ‘early registrations’ to the event Francis, Paul Henschke, Michael Esler, Bob Industry Technical Conference will be provided by 8 June 2001. Dambergs and Miguel de Barros Lopes. Peter in the 2002 Annual Report. However, Godden (and his team), Don Buick and Sally during the year, Rae Blair, in her role as The Institute’s Director, Peter Høj is Chair of Bell maintain the ‘Services’ section. Rae Blair Conference Manager and Treasurer of the Conference Planning Committee; Peter maintains the ‘About Us’ section, and Creina Australian Wine Industry Technical Conference Godden, Trudy Goode and Ella Robinson are Stockley is responsible for the ‘Information’ Inc.; spent much time liaising with coordinating 74 workshops and Jeremy Hack section. Dr Alan Pollnitz undertakes the international speakers, venue operators, is coordinating the poster display. Excellent actual updating of the www files in his role sponsors, graphic designers and exhibition backup is provided by our Accountant and as part-time Computer Systems Officer. management. She is ably assisted by the Company Secretary. Secretariat of the Conference, Carolyn Grant,

49 Team Reports

11AWITC website workload particularly from the McLaren Vale The sensory assessment service has been a During the year, the Conference Manager region has occurred during the 2001 vintage significant growth area with large numbers purchased server space and coordinated the period. This is evident from the increased of samples assessed for taints, faults or production and uploading of information for numbers of tests for malic acid performed general quality parameters. New software the website of the Australian Wine Industry during the year. The personal contact with purchased by the Institute has streamlined Technical Conference. The website address winemakers, technical staff and export staff the recording of comments, the scoring and is www.awitc.com.au. has been successful and will be continued in the reporting of sensory work. Much of the the new year. For the first time, the sensory work has been conducted in 26th World Congress and 81st General Analytical Service sponsored a trophy for ‘the conjunction with chemical testing and used Assembly of the Office International de best tawny port’ at the Adelaide Wine show for dispute resolution or insurance purposes. la Vigne et du Vin in September 2000. The Communication and Publicity Manager Staff have worked closely with the Industry also manages the Secretariat of the OIV In October 2000, a new fee schedule Services’ team to analyse the 18 month and Congress, Marcia Burnett. The Institute incorporating details and pricing for the 24 month samples from the closure trial (see Director and the Communication and services offered was mailed to all clients. report under Industry Services activities). The Publicity Manager are both members of the The terms and conditions of our services analysis of large numbers of free and total Australian OIV Planning Committee. The have been brought to the attention of clients sulphur dioxide, dissolved oxygen and Communication and Publicity Manager secured through the fee schedule, the web site and OD420 were performed with staff working accommodation for delegates; produced the in formal quotations. Formal quotations are in shifts to complete the work within two to logo, letterhead, first announcement and issued for larger or non-routine jobs three days of commencement. poster announcing the event in Australia, including clear identification of the services the preliminary programme (in three to be provided. Most incoming work of a John Hughes successfully completed the languages) and established a skeleton routine nature has client requirements Advanced Wine Assessment Course conducted budget for the event. Further details of the identified by letter or sample labels. in July 2000 and has contributed to the Institute’s contribution to the OIV Congress planning of the July 2001 course to be and General Assembly held in Australia will The services provided include analyses conducted by Industry Services. Other be shown in the 2002 Annual Report. consistent with the requirements of export Analytical Service staff contributed to the course certification and support for the quality control including pouring of wine, cleaning up and activities of winemaking and viticulture often recording of the trainee’s results for evaluation. Analytical Service requiring more specialised skills than those commonly found in production laboratories. The Analytical Service was re-assessed by Staff: Don Buick, John Hughes, Matthew The Analytical Service conducted over NATA in September of 2000 and complimented Holdstock, Greg Ruediger, Gayle 46,000 individual analyses on wine and on the quality of its documentation and Baldock, Amanda Cook, Matthew grapes in 2000/2001 and covering simple procedures. The method for ochratoxin A Cream, Jeremy Hack, Radka Kalouch, analyses such as malic acid to more complex was approved and the inclusion of malathion Andrea Kemp, Adam Loveys, Kevin analyses such as the multi-residue screen for and fenitrothion in the multiresidue assay is Pardon, Randall Taylor, Sandra Lloyd- agricultural chemicals. The aim is to provide under review. Don Buick and Randell Taylor Davies, Bao Tran and David Boehm a range of quality analytical services that are were approved as NATA signatories. NATA important to industry and which can be accreditation previously required compliance The Analytical Service is a commercial facility provided in a reliable, price competitive, with ISO Guide 25. This international operating independently on a fully-costed accurate and timely manner. standard has been replaced by ISO17025 basis within the Institute with the Manager, with laboratories required to meet the new Mr Don Buick, reporting to the Director. It is The number of tests performed by the standard by 2002. Compliance with funded by the income generated from fee Analytical Service increased by 3% from the ISO17025 ensures the laboratory operates in earning work and contributes significantly to previous year with significant increases in some accordance with ISO9000. The assessment the financial well being of the Institute. The areas of testing being offset by a lower provided a check against the new standard Analytical Service provides services to the number of tests performed as part of the and confirmed good progress towards Australian wine industry including use of the Institute led closure trial during the year. compliance with the new standard. practical outcomes of the Institute’s Significant increases occurred in AWBC published research. Due to the nature of certificates, EU certificates, malic acid, sensory Trace Analysis Laboratory some of its work, the Analytical Service, assessments, multi-residue assays, colour in The Trace Analysis Laboratory, operating under appropriate cross-charging red grapes, glycosyl-glucose in grapes and 4- within the Analytical Service, performs more arrangements, makes use of the expertise ethyl phenol analyses. The number of data specialised testing including a multi-residue existing within the Industry Service team and points produced increased by 7% providing screen using gas chromatography/mass within the Research groups to interpret or a better measure of the increase in workload. spectrometry including 30 residues, covering advise on results or project design. over 50 brand name agrochemicals. Several Analytical methods for organic acids, pesticides given special approval for use in Don Buick and John Hughes have visited ascorbic acid, and methanol have been re- controlling plague locusts have been clients in the McLaren Vale region and in the validated following the purchase of a new included in the multi-residue screen Margaret River region during the year as well high pressure liquid chromatograph and the including chlorpyrifos, carbaryl, dimethoate, as attending Wine Australia in Melbourne in refurbishment of a gas chromatograph. malathion and fenitrothion. A multi-residue November and the Margaret River Field Day screen for several compounds unable to be in May. These visits plus direct mail and the placement of advertisements in industry journals and publications have assisted in the 97/98 98/99 99/00 00/01 development of improved client relationships Total number of tests 35852 39087 44846 46037 and the generation of several new clients. As a consequence, a significant increase in Table 10: Comparison of selected tests performed by the Analytical service by period.

50 Greg Ruediger, Don Buick and John Hughes analysed by gas chromatography has been samples indicates that industry practitioners developed using high pressure liquid seriously should consider performing this chromatography and includes cyprodinil, relatively cheap test on a more routine basis. fludioxinil, pyrimethanil, fenhexamid and tebufenozide. Other methods have been The validation of a method for the developed and validated for spiroxamine, determination of histamine in wine using trifloxystrobin, axoystrobin and indoxacarb. high pressure liquid chromatography has commenced following limited success using Several extended residue trials have been an ELISA kit method for this purpose. The performed for chemical companies during measurement of histamine is necessary to the year to establish the levels of residue meet the requirements for importation into present or absent in grapes or wine Switzerland and will allow a survey of Australian following the application to vines of newly wine to be performed in the new year. developed agrochemicals. These trials Little change in staffing occurred during the involve testing of grapes, small scale year. Adam Loveys formerly employed as a winemaking and analysis following casual Laboratory Technician graduated from development and validation of methods of the University of Adelaide with an Honours analysis. The data produced is used by the Degree in Agricultural Science and chemical companies for further development subsequently resigned to take up a of agrochemicals and for application to the viticultural position on Kangaroo Island. National Registration Authority for Adam has worked at the Institute for several registration approval. From 2003 this work years and made a valuable contribution to will require Good Laboratory Practice (GLP) the Analytical Service whilst continuing his accreditation from NATA. The Trace Analysis studies. John Hughes took long service Laboratory has begun to prepare for leave during the year including travelling in accreditation and Greg Ruediger is to attend the winemaking regions of Germany and a training course on GLP in the near future. France. Matt Holdstock, Analytical Service Supervisor, has commenced part-time studies Large numbers of samples have been analysed to complete the Graduate Diploma in during the year for chloroanisoles, the principle Oenology. This will mean that the Analytical contributors to cork taint in wine and for 4- Service will have two degree qualified ethylphenol which is an indicator of oenologists able to service industry’s needs. Brettanoyces/Dekkera yeast spoilage activity. Several other staff have attended specialised 4-ethylphenol is included in the oak flavour instrument training courses to ensure new compound assay but is now offered separately equipment is well maintained and used to its when only 4-ethylphenol is required. maximum capability. The Analytical Service Eugenol is now included in the oak flavour aims to support the continual upgrading of compounds assay. The relatively high staff skills as an important element in concentrations of 4-ethylphenol in many expanding our portfolio of services.

51 Statement of Financial Performance

For the year ended 30 June 2001 2001 2000 $ $ Revenue from operating activities

Grape and Wine Research and Development Corporation Project funds 3,569,186 3,191,013 Capital grants Building 0 250,000 Equipment 426,000 722,514

Other project funds 545,348 409,742

Analytical Service 1,167,825 1,130,203

Sundry income 193,853 161,180

Expenses from operating activities

Employee benefit expense 3,437,532 2,961,477

Consumables used 514,417 418,478

Depreciation and amortisation expense 551,179 338,130

Repairs and maintenance 99,282 72,429

Travel expenses 96,512 98,800

Borrowing cost expense 12,406 24,447 Hans Muhlack

Other expenses from ordinary activities 959,196 1,092,163

Profit from operating activities 231,688 883,175

Net gain (loss) on disposal of assets Motor vehicles 14,935 0 Equipment 397 (495)

Profit from ordinary activities 247,020 882,680

Write back of asset revaluation reserve on reversion to measuring buildings at cost (603,067) 0

Transfer of accumulated amortisation on buildings 125,536 0

Total changes in equity (230,511) 882,680

52 Statement of Financial Position

As at 30 June 2001 2001 2000 $ $ Current assets

Cash assets 920,798 120,020

Receivables 307,584 346,413

Commercial bills 670,000 648,000

Other current assets 52,326 25,621

Total current assets 1,950,708 1,140,054

Non current assets

Leasehold buildings 1,596,247 2,102,021

Plant and equipment 1,918,127 1,960,983

Australian Wine Industry Chair of Oenology 840,000 840,000

Total non current assets 4,354,374 4,903,004

Total Assets 6,305,082 6,043,058

Current liabilities

Payables and other accruals 924,288 436,328

Project funds not expended

GWRDC 37,150 51,877 CRCV 13,936 0

Interest bearing liabilities 120,000 170,000

Provisions 391,490 362,822

Total current liabilities 1,486,864 1,021,027

Non current liabilities

Provisions 90,970 64,272

Total non current liabilities 90,970 64,272

Total Liabilities 1,577,834 1,085,299

Net Assets 4,727,248 4,957,759

Equity

Reserves 966,750 1,569,817

Retained profits 3,760,498 3,387,942

Total Equity 4,727,248 4,957,759

53 Statement of Cash Flows

For the year ended 30 June 2001 2001 2000 $ $

Cash flows from operating activities

Grants and other income 5,874,426 5,929,599

Interest received 68,845 67,825

Payments to suppliers and employees (4,605,745) (4,701,320)

Net cash provided by operating activities 1,337,526 1,296,104

Cash flows from investing activities

Payment for commercial bills (22,000) (27,000)

Payments for building improvements (8,858) (684,620)

Payments for plant and equipment (507,390) (911,819)

Proceeds from sale of plant and equipment 51,500 49,283

Net cash used in investing activities (486748) (1,574,156)

Cash flows from financing activities

Repayment of loans (50,000) 0

Proceeds from loans 0 170,000

Net cash provided by financing activities (50,000) 170,000

Net increase (decrease) in cash held 800,778 (108,052)

Cash at 1 July 120,020 228,072

Cash at 30 June 920,798 120,020

Reconciliation of net cash provided by ordinary activities with ordinary profit

Profit from ordinary activities 247,020 882,680

Non cash flows in operating profit

Amortisation and depreciation 551,179 338,130 (Profit) loss on the sale of plant and equipment (15,332) 495 Charges to (reduction in) provisions 55,366 45,351

Changes in assets and liabilities

(Increase) decrease in inventories (29,332) (12,000) (Increase) decrease in receivables and prepayments 41,456 (68,043) Increase (decrease) in sundry creditors and accruals 487,169 109,491

Net cash provided by ordinary activities 1,337,526 1,296,104

54 Appendix

Appendix 1. External seminars, talks and poster papers presented by institute staff during 2000/01

Author Title Organization/Location Date

External seminars and talks

K.F. Pocock, Effects of winemaking and Lincoln University Grape and Wine 24 July 2000 D. Tattersall2, viticultural practices on protein School, Christchurch, New Zealand Y. Hayasaka, instability in wine I. Dupin1,2, B. McKinnon1,2, R. van Heeswijck2, P. H øj, E.J. Waters

Z.K. Peng, P.J. Iland2, The influence of winemaking Lincoln University Grape and Wine 24 July 2000 R. Ristic2, R. Gawel2,5, and viticultural practices on School, Christchurch, New Zealand A. Oberholster1,2, colour and tannins in red wine I.L. Francis, E.J. Waters

P.W. Godden Bunch rots: understanding the ASVO Managing Bunch Rots seminar Mildura Vic 28 July 2000 winemaker's dilemma

P.W. Godden Persistent wine instability issues, New Horizons for Wine Quality, Victorian Wine 10 August 2000 and, Towards better barrel Industry Association’s Seminar. Melbourne Vic management – a review of the AWRI oak research

P.B. Høj What happened in the wine Committee for Economic Development of Australia’s 6 September 2000 industry in the last 10 years that seminar ‘Building of the knowledge economy’ has transformed it from a shadow Hyatt Regency Adelaide of Europe to the vibrant player in the global area it is today?

G.M. Elsey C13-Norisoprenoids: their generation Flinders University weekly seminar 8 September 2000 and contribution to aroma program, Flinders University, SA

P.A. Henschke Fermentation management and E. & J. Gallo Winery, Sonoma, 10 September 2000 importance of yeast nutrition California, USA

P.B. Høj Genetic engineering, the First National Environmental Conference Adelaide SA 1 November 2000 environment and the wine industry - a personal perspective

P.W. Godden, Roadshow seminars McLaren Vale, SA 8 November 2000 P.J. Costello, S.J. Bell, E.J. Bartowsky, A.D. Coulter

P.B. Høj Current trends in Australian South African Society for Enology and Viticulture’s 8-10 November 2000 oenological research: the quest 2nd International Viticulture and Enology Congress, to produce grapes and wine to Cape Town South Africa a predetermined specification

P.W. Godden, Roadshow seminars Adelaide Hills, SA 9 November 2000 S.J. Bell, M. Gishen, I.L. Francis, G.A. Ruediger, A.D. Coulter

P. B . H øj, P.W. Roadshow seminars Barossa Valley, SA 13 November 2000 Godden, S.J. Bell, M.A. Sefton, E.J. Bartowsky, P.A. Henschke, I.L. Francis, M. Gishen

55 Appendix

Appendix 1. External seminars, talks and poster papers presented by institute staff during 2000/01

Author Title Organization/Location Date

P. B . H øj, P.W. Roadshow seminars Clare Valley SA 16 November 2000 Godden, S.J. Bell, M.A. Sefton, E.J. Bartowsky, P.A. Henschke, I.L. Francis, M. Gishen

P.B. Høj An update of the use of Near Grape and Wine Industry Outreach 17 November 2000 Infrared Spectroscopy for Seminar, Adelaide University, Plant measurement in grape quality Research Centre, Urrbrae SA parameters

P.J. Costello, Towards understanding the Grape and Wine Industry Outreach 17 November 2000 P.A. Henschke, interactions between wine yeast Seminar, Adelaide University, Plant A.J. Markides and lactic acid bacteria Research Centre, Urrbrae SA

P.A. Henschke Wine microbiology and Lallemand meeting, AWRI Urrbrae, SA 20 November 2000 biotechnology projects overview

J.M. Eglinton The search for alternative wine Lallemand meeting, AWRI Urrbrae, SA 20 November 2000 yeasts. Flavour and aroma diversity using Saccharomyces bayanus

E.J. Bartowsky Microbial spoilage of bottled red Lallemand meeting, AWRI Urrbrae, SA 20 November 2000 wine by acetic acid bacteria, and, Diacetyl ‘buttery’ attribute of wine and MLF?

P.J. Costello Mousy off-flavour spoilage of Lallemand meeting, AWRI Urrbrae, SA 20 November 2000 wine, and, Interaction of wine yeast and lactic acid bacteria

M. Esler New technologies for rapid Interwinery Analysis Group meeting, 24 November 2000 measurement (NIRS and FT-IR) Renmark, SA

P. B . H øj, P.W. Roadshow seminars Berri, SA 4 December 2000 Godden, S.J. Bell, P.A. Henschke, I.L. Francis, M. Gishen

M.A. Sefton, M. Precursors to oak lactone: Annual Adelaide Organic Chemistry 4 December 2000 Raunkjaer, G.K. synthesis of gallate ester Symposium, Flinders University, SA Skouroumounis, derivates of 3-methyl-4- G.M. Elsey hydroxyoctanoic acid

P.W. Godden, S.J. Roadshow seminars Mildura, Vic 6 December 2000 Bell, P.A. Henschke, M. Gishen, A.D. Coulter

I.L. Francis Tannins and mouthfeel Southcorp Winemakers’ Forum Mildura, Vic 7 December 2000

P. B . H øj, P.W. Roadshow seminars Geelong, Vic 15 December 2000 Godden, A.D. Coulter

P.A. Henschke Brettanomyces - winemaking Southern Pinot Noir Workshop, 18 January 2001 significance and prevention Methven, Canterbury, New Zealand

A.P. Pollnitz, The analysis of wine volatiles Australian and New Zealand Society 5 February 2001 G.K. Skouroumounis, derived from oak for Mass Spectrometry, 18th Conference, K.H. Pardon, D. Capone, Coolangatta, Qld M.A. Sefton

56 Appendix 1. External seminars, talks and poster papers presented by institute staff during 2000/01

Author Title Organization/Location Date

Y. Hayasaka, K.S. Use of electrospray mass Australian and New Zealand Society for Mass 6 February 2001 Adams2, K.F. spectrometry for mass Spectrometry, 18th Conference, Coolangatta, Qld Pocock, G.A. determination of grape ( Baldock, E.J. Waters, vinifera) juice pathogenesis P.B. Høj related proteins: potential tool for varietal differentiation

E.J. Bartowsky Wine microbiology Non-clinical meeting of the Australian 28 March 2001 Society for Microbiology, Plant Research Centre, Urrbrae, SA

I.L. Francis Relating grape composition and Grape Expectations: University of 11 April 2001 wine quality, and, Key grape- California, Davis, California, USA derived wine flavour compounds: can we define flavour specifications?

C.S. Stockley A role for wine in the diet Department of Clinical Dietetics’ continuing education 11 April 2001 program, The Royal Adelaide Hospital

I.L. Francis Near infrared for rapid grape analysis Grape Expectations: University of California, 12 April 2001 Davis, California, USA

E.J. Waters Program 1: Vineyard management Cooperative Research Centre Board, 9 May 2001 to meet grape quality specifications Plant Research Centre, Urrbrae, SA

M.J. Herderich Winegrape tannin and CRCV Second Year Review, 22 May 2001 colour specification Plant Research Centre, Urrbrae, SA

P.B. Høj Advice to young scientists, a Adelaide University Science Scholars 29 May 2001 personal perspective

I.L. Francis Wine flavour and sensory analysis Australian Institute of Food Science and Technology 30 May 2001 (Victoria) Sensory Evaluation Group, Werribee, Vic

P.B. Høj Use of research outcomes for BRL Hardy Vintage Conference, 4 June 2001 the management of problems Lake Crackenback, NSW and opportunities

E.J. Waters, K.F. Pocock, The unstable proteins of wine: Unité Mixte de Recherche Sciences pour l'Oenologie, 13 June 2001 D. Tattersall2, Y. Hayasaka, a summary of their properties INRA, Montpellier, France I. Dupin1,2, B. McKinnon1,2, and of factors influencing the R. van Heeswijck2, P.B. Høj1,2 levels of these

P.B. Høj A general introduction to The Unité Mixte de Recherche Sciences pour l'Oenologie, 13 June 2001 Australian Wine Research INRA, Montpellier, France Institute and an outline of selected research activities

S. Vidal Sensory features of grape tannins In Vino Analytica Scientia, Bordeaux, France 15 June 2001

E.J. Waters, K.F. Pocock, The unstable proteins of wine: In Vino Analytica Scientia, Bordeaux, France 15 June 2001 D. Tattersall2, Y. Hayasaka, a summary of their properties I. Dupin1,2, B. McKinnon1,2, and of factors influencing the R. van Heeswijck2, P.B. Høj1,2 levels of these proteins in wine

M.J. Herderich Red wine colour and mouthfeel: American Society of Enology and Viticulture 29 June 2001 analysis of phenolic pigments Annual General Meeting, San Diego, USA and tannins

57 Appendix

Appendix 1. External seminars, talks and poster papers presented by institute staff during 2000/01

Author Title Organization/Location Date

Posters

M.B. Esler, M. Effects of variety, region and 10th International Conference on Near Infrared 10-15 June 2000 Gishen, I.L. Francis, season on near infrared Spectroscopy, Kyongju, South Korea R.G. Dambergs, A. reflectance spectroscopic Kambouris, W.U. analysis of quality parameters Cynkar, D.R. Boehm in red wine grapes

R.G. Dambergs, Wine quality grading by near 10th International Conference on Near Infrared 10-15 June 2000 A. Kambouris, N. infrared spectroscopy Spectroscopy, Kyongju, South Korea Schumacher, I.L. Francis, M.B. Esler, M. Gishen

Workshops

S-J. Bell Spray application in viticulture Research to Practice™, McLaren Vale, SA 15 August 2000

S-J. Bell Spray application in viticulture Research to Practice™, McLaren Flat, SA 17 August 2000

S-J. Bell Grapevine nutrition Research to Practice™, Langhorne Creek, SA 26 September 2000

S-J. Bell Integrated Pest Management Research to Practice™, McLaren Vale, SA 7 November 2000

P.W. Godden, A.D. Identification and management McLaren Vale, SA 8 November 2000 Coulter, P. Valente, of wine instabilities E.M. Robinson, M. Gishen

S-J. Bell Grapevine Nutrition Research to Practice™, Barossa Valley, SA 11 November 2000

S-J. Bell Grapevine Nutrition Research to Practice™, McLaren Vale, SA 13 November 2000

P.W. Godden, A.D. Identification and management Barossa Valley, SA 14 November 2000 Coulter, P. Valente, of wine instabilities E.M. Robinson

P.W. Godden, A.D. Identification and management Clare Valley, SA 17 November 2000 Coulter, P. Valente, of wine instabilities E.M. Robinson

P.W. Godden, A.D. Identification and management Berri, SA 5 December 2000 Coulter, P. Valente, of wine instabilities E.M. Robinson

P.W. Godden, A.D. Identification and management Mildura, Vic 7 December 2000 Coulter, P. Valente, of wine instabilities E.M. Robinson

Advanced Wine Assessment Course

P.W. Godden, E. Robinson Adelaide SA 18-21 July 2000 J.B. Hughes, M. Gishen, A. Loveys, A.D. Coulter I.L. Francis

58 Appendix 2. Teaching responsibilities of Institute staff during 2000/01

Subject Number of lectures Institute staff

2000—Semester 2

Adelaide University 9685 Advances in Oenology 2 Dr M.A. Sefton 9685 Advances in Oenology 2 Dr I.L. Francis 9685 Advances in Oenology/ 9086 Advances in Oenology (supplemented) 3 Dr P.A. Henschke 9685 Advances in Oenology/ 9086 Advances in Oenology (supplemented) 2 Dr E.J. Bartowsky 9685 Advances in Oenology/ 9086 Advances in Oenology (supplemented) 1 Dr P.J. Costello 9685 Advances in Oenology 1 P.W. Godden 1958 Wine packaging and quality management 2 C.S. Stockley 1958 Wine packaging and quality management 1 M. Gishen

The Flinders University of South Australia MMED 3921 Industrial and Pharmaceutical Microbiology 1 Dr P.A. Henschke

2001—Semester 1

Adelaide University 2580 Stabilisation and Clarification 3 Dr E.J. Waters 2580 Stabilisation and Clarification 1 P.W. Godden 1005/3113 Winemaking 2 Dr P.A. Henschke 1958 Wine Packaging and Quality Management 1 M. Gishen 2213 Grape Industry Practice Policy and Communication ~ 40 hours Professor P.B. Høj and (subject coordination and delivery for five weeks) C.S. Stockley 5693 Wine and Marketing in Society 6 C.S. Stockley

59 Appendix

Appendix 3. Graduate and Honours student supervision responsibilities of Institute staff for 2000/01

Student Supervisor/s Source of funds

Honours

C. Day P.B. Høj, S.J. Clarke2 Adelaide University A. Matthews E.J. Bartowsky, I.L. Francis Adelaide University M. Mercurio G.K. Skouroumounis, G.M. Elsey Flinders University M.A. Smith G.M. Elsey Flinders University A. Walkenhorst I.L. Francis, R.G. Dambergs Adelaide University

Masters

D. Coates E.J. Bartowsky, R. Gawel2,5 Adelaide University

PhD S.L. Brown M.A. de Barros Lopes, E.J. Waters, P.B. Høj GWRDC

F. Carrau P.A. Henschke, E. Dellacassa8 University of the Republic of Uruguay staff member N. D’Incecco E.J. Bartowsky, P.A. Henschke Italian PhD Scholarship

J.M. Eglinton P.A. Henschke, P.R. Langridge6 Institute staff

R. Gawel I.L. Francis, A.J. Markides2,4 DHVO staff

W. Greenrod M. Fenech9, M.Abbey9, M.A. Sefton, P. Burcham10 GWRDC S. Hoffmann P. Winterhalter, C.S. Stockley University of Braunschweig

K.S. Howell P.A. Henschke, E.J. Bartowsky, G.H. Fleet3 APRA scholarship

A. Janusz G.M. Elsey, M.A. Sefton, M. Perkins7 CRCV

D. Lee G.P. Jones2, E.J. Waters CRCV

A. Oberholster E.J. Waters, I.L. Francis, G.P. Jones2, P.G. Iland2 GWRDC

C.J. Puglisi G.M. Elsey, M.A. Sefton, R. Prager7 CRCV R. Ristic P.J. Iland, I.L. Francis Adelaide University P. Sivilotti S.J. Bell Italian Government and University of Udine H. Smyth (nee Pain) I.L. Francis, M.A. Sefton GWRDC

K.L. Wilkinson M.A. Sefton, R. Prager7, G.M. Elsey GWRDC

60 Theses completed

Student PhD/Honours Title Supervisor/s

Robert Asenstorfer PhD Isolation, structures and properties of Dr G.P. Jones2, Dr P.J. Iland2, Dr E.J. Waters anthocyanins and oligmeric pigments in wine

Patrick Iland PhD A study of glycosides in grapes and wines Dr B. Coombe2, Dr P.J. Williams, Professor P.B. Høj

Patrick Jones PhD Glucosyl transferases involved in the biosynthesis Professor P.B. Høj, Dr B.L. Møller11 of cyanogenic glucosides in sorghum bicolor

Alan Pollnitz PhD The analysis of volatile wine components derived Dr M.A. Sefton, Dr G.P. Jones2 from oak products during winemaking and storage

Christopher Smyl PhD L-Proline: a potential nitrogen source for yeast Dr P.A. Henschke, Professor P. Langridge6, Saccharomyces cerevisiae under anaerobic conditions Dr M.A. de Barros Lopes

Vanessa Stockdale PhD Biochemical analysis of mannoproteins associated Dr E.J. Waters, Dr P.J. Williams, with haze protection in white wine Dr G.B. Fincher6

Nicholas Yap PhD The sensitivity of yeasts to killer yeast toxin, Dr P.A. Henschke, Dr M.A. de Barros Lopes, with focus on the killer yeast Pichia membranifaciens Professor P. Langridge6

Julie Campbell Honours The influence of chip size, heating processes Dr A.P. Pollnitz, Dr M.A. Sefton and other factors on the impact of oak chips on wine composition

David Dowden Honours Effect of polysaccharides on wine instability Dr G.P. Jones2, Dr E.J. Waters

Freya Hohnen Honours Changes in seed phenolic composition during berry Dr P.J. Iland2, Dr Z.K. Peng maturation and throughout fermentation/extended maceration of Vitis vinifera cv. Cabernet Sauvignon

Helen McCarthy Honours Evaluation of the ability of the chemical sensory Dr E.J. Bartowsky, Dr I.L. Francis nose to determine the aroma profiles of wines which have undergone malolactic fermentation

Marie Pearce Honours Optimising diacetyl levels in wine Dr E.J. Bartowsky, Dr A.J. Markides2,4

Nina Viergutz Honours The interaction between wine yeast and Dr P.J. Costello, Dr A.J. Markides2,4 lactic acid bacteria

1 The Australian Wine Research Institute, 2 Department of Horticulture, Viticulture and Oenology, Adelaide University; 3 Department of Food Science, University of New South Wales; 4 now with Lallemand Australia; 5 now with Roseworthy Programs Pty Ltd; 6 Department of Plant Science, Adelaide University; 7 Flinders University; 8 University of the Republic of Uruguay, 9 CSIRO Health Sciences and Nutrition, 10 Department of Clinical and Experimental Pharmacology, Adelaide University, 11 Royal Agricultural University of Copenhagen

61 Appendix

Appendix 4. Institute staff publications 2000/01

629 Carrau, F.M.; Henschke, P.A.; Medina, K.; Gioia, O.; Dellacassa, E. Assimilable nitrogen addition can cause stuck wine fermentations with mixed cultures of Saccharomyces cerevisiae killer and sensitive yeast. VII Congreso Latino Americano de viticultura y enología; 28 de Noviembre al 3 de Diciembre de 1999; Mendoza, Argentina: Instituto Nacional de Vitivinicultura; 2000: 149–154. 630 Skouroumounis, G.K.; Sefton, M.A. Acid-catalyzed hydrolysis of alcohols and their ß-D-glucopyranosides. J. Agric. Food Chem. 48: 2033–2039; 2000. 631 Waters, E.J.; Dupin, I.; Stockdale, V. A review of current knowledge on polysaccharides which ‘protect’ against protein haze in white wine. Aust. Grapegrower Winemaker (438a): 13–16; 2000. 632 Eglinton, J.; Fogarty, M.; McWilliam, S.; Francis, L.; Kwiatkowski, M.; Høj, P.; Henschke, P. Using Saccharomyces bayanus to modify the chemical and sensory profile of wine. Aust. Grapegrower Winemaker (438a): 28–32; 2000. 633 Todd, B.E.N.; Fleet, G.H.; Henschke, P.A. Promotion of autolysis through the interaction of killer and sensitive yeasts: potential application in sparkling wine production. Am. J. Enol. Vitic. 51: 65–72; 2000. 634 Stockley, C.S. Ochratoxin A–a metabolite on the agenda for the global wine industry. Aust. Grapegrower Winemaker (438a): 111–112; 2000. 635 Stockley, C.S. Advances in Australian research into the potential cardioprotective properties of wine. XXVth World Congress of the OIV; 19–23 June 2000; Paris, France: Office International de la Vigne et du Vin; 2000: 33–41. 636 Gishen, M.; Holdstock, M. Preliminary evaluation of the performance of the Foss WineScan FT120 instrument for the simultaneous determination of several wine analyses. Aust. Grapegrower Winemaker (438a): 75–81; 2000. 637 Bartowsky, E.J.; Henschke, P.A. Management of malolactic fermentation for the ‘buttery’ diacetyl flavour in wine. Aust. Grapegrower Winemaker (438a): 58–67; 2000. 638 Pollnitz, A.P.; Pardon, K.H.; Sefton, M.A. 4-ethylphenol, 4-ethylguaiacol and oak lactones in Australian red wines. Aust. Grapegrower Winemaker (438): 45–52; 2000. 639 Dupin, I.V.S.; McKinnon, B.M.; Ryan, C.; Boulay, M.; Markides, A.J.; Jones, G.P.; Williams, P.J.; Waters, E.J. Saccharomyces cerevisiae mannoproteins that protect wine from protein haze: their release during fermentation and lees contact and a proposal for their mechanism of action. J. Agric. Food Chem. 48: 3098–3105; 2000. 640 Haselgrove, L.; Botting, D.; van Heeswijck, R.; Høj, P.B.; Dry, P.R.; Ford, C.; Iland, P.G. Canopy microclimate and berry composition: the effect of bunch exposure on the phenolic composition of Vitis vinifera L cv. Shiraz grape berries. Aust. J. Grape Wine Res. 6: 141–149; 2000. 641 Stines, A.P.; Grubb, J.; Gockowiak, H.; Henschke, P.A.; Høj, P.B.; van Heeswijck, R. Proline and arginine accumulation in developing berries of Vitis vinifera L. in Australian vineyards: influence of vine cultivar, berry maturity and tissue type. Aust. J. Grape Wine Res. 6: 150–158; 2000. 642 Godden, P. The use of copper sulphate in winemaking. Aust. N.Z. Wine Ind. J. 15: 66–67; 2000. 643 Asenstorfer, R.E.; Hayasaka, Y.; Iland, P.G.; Lambert, S.G.; Jones, G.P. Wine phenolics: the development of pigments in red wine. Steans, G., ed. Proceedings of the 1999 conference of the New Zealand Society for Viticulture and Oenology; 4-5 November 1999; Auckland, New Zealand Society for Viticulture and Oenology; 1999: 83-78. 644 Yap, N.A.; de Barros Lopes, M.; Langridge, P.; Henschke, P.A. The incidence of killer activity of non-Saccharomyces yeasts towards indigenous yeast species of grape must: potential application in wine fermentation. J. Appl. Microbiol. 89: 381–389; 2000. 645 Godden, P. Persistent wine instability issues. Aust. Grapegrower Winemaker (443): 10–14; 2000. 646 West, S.; Bell, S.-J.; Henry, K. Some facts about locust control in vineyards. Aust. Grapegrower Winemaker (443): 20–22; 2000. 647 Eglinton, J.M.; McWilliam, S.J.; Fogarty, M.W.; Francis, I.L.; Kwiatkowski, M.J.; Høj, P.B.; Henschke, P.A. The effect of Saccharomyces bayanus mediated fermentation on the chemical composition and aroma profile of Chardonnay wine. Aust. J. Grape Wine Res. 6: 190–196; 2000. 648 Gawel, R.; Oberholster, A.; Francis, I.L. A ‘Mouth-feel Wheel’: terminology for communicating the mouth-feel characteristics of red wine. Aust. J. Grape Wine Res. 6: 203–207; 2000. 649 de Barros Lopes, M.; Rehman, A.-U.; Gockowiak, H.; Heinrich, A.J.; Langridge, P.; Henschke, P.A. Fermentation properties of a wine yeast over-expressing the Saccharomyces cerevisiae glycerol 3-phosphate dehydrogenase gene (GPD2). Aust. J. Grape Wine Res. 6: 208–215; 2000. 650 Grbin, P.R.; Henschke, P.A. Mousy off-flavour production in grape juice and wine by Dekkera and Brettanomyces yeasts. Aust. J. Grape Wine Res. 6: 255–262; 2000. 651 Gishen, M.; Dambergs, R.G.; Kambouris, A.; Kwiatkowski, M.; Cynkar, W.U.; Høj, P.B.; Francis, I.L. Application of near infrared spectroscopy for quality assessment of grapes, wine and spirits. Davies, A.M.C.; Giangiacomo, R., eds. Near Infrared Spectroscopy: proceedings of the 9th international conference; Chichester, UK. NIR Publications; 2000: 917–920. 652 Ganter, P.F.; de Barros Lopes, M. The use of anonymous DNA markers in assessing worldwide relatedness in the yeast species Pichia kluyveri Bedford and Kudrjavzev. Can. J. Microbiol. 46: 967–980; 2000. 653 Stockley, C. A review of recent findings on the relationship between wine and health. Aust. N.Z. Wine Ind. J. 15: 36–43; 2000. 654 Godden, P. Bunch rots–understanding the winemaker’s dilemma. Davies, C.; Dundon, C.; Johnstone, R., eds. Managing bunch rots: proceedings of a seminar; 28 July 2000, Mildura Arts Centre, Mildura, Victoria. Adelaide, S.A.: Australian Society of Viticulture and Oenology; 2000: 52–54. 655 Jones, P.R.; Andersen, M.D.; Nielsen, J.S.; Høj, P.B.; Møller, B.L. The biosynthesis, degradation, transport and possible function of cyanogenic glucosides. Romeo, J.T.; Ibrahim, R.; Varin, L.; de Luca, V., eds. Evolution of metabolic pathways. Oxford: Pergamon; 2000: 191–247. (Recent advances in phytochemistry; 34).

62 656 Gawel, R.; Iland, P.G.; Francis, I.L. Characterizing the astringency of red wine: a case study. Food Qual. Pref. 12: 83–94; 2001. 657 Eglinton, J.; Henschke, P.A. The effect of a high concentration of acetic acid on the restarting of a stuck ferment. Aust. N.Z. Wine Ind. J. 16: 77–81; 2001. 658 Stockley, C.S. Evidence of the specific benefits of wine consumption on human health – an update on the cardiovascular and other physiological mechanisms. Aust. Grapegrower Winemaker (446): 72–77; 2001. 659 Stockley, C.S. Advances in Australian research into the potential cardioprotective properties of wine. Bull. OIV 73: 660–675; 2000. 660 Peng, Z.; Hayasaka, Y.; Iland, P.G.; Sefton, M.; Høj, P.; Waters, E.J. Quantitative analysis of polymeric procyanidins (tannins) from grape (Vitis vinifera) seeds by reverse phase high-performance liquid chromatography. J. Agric. Food Chem. 49: 26–31; 2001. 661 Stewart, R.J; Varghese, J.N; Garrett, T.P.J; Høj, P.B.; Fincher, G.B. Mutant barley (1,3;1,4)-ß-D-glucan endohydrolases with enhanced thermostability. Protein Engineering 14: 245-253; 2001. 662 Barker, D.A.; Capone, D.L.; Pollnitz, A.P.; McLean, H.J.; Francis, I.L.; Oakey, H.; Sefton, M.A. Absorption of 2,4,6-trichloroanisole by wine corks via the vapour phase in an enclosed environment. Aust. J. Grape Wine Res. 7:40-46; 2001. 663 Hayasaka, Y.; Adams, K.S.; Pocock, K.F.; Baldock, G.A.; Waters, E.J.; Høj, P.B. Use of electrospray mass spectrometry for mass determination of grape (Vitis vinifera) juice pathogenesis-related proteins: a potential tool for varietal differentiation. J. Agric. Food Chem. 49:1830-1839; 2001. 664 Williams, P.J.; Francis, I.L. Wine flavor research-experience from the past offer a guide to the future. Rantz, J.M., ed. Proceedings of the ASEV 50th Anniversary Annual Meeting; June 19-23, 2000; Davis, California. American Society for Enology and Viticulture; 2001: 191-195. 665 Høj, P.B.; Tattersall, D.B.; Adams, K.; Pocock, K.F.; Hayasaka, Y.; van Heeswijck, R.; Waters, E.J. The ‘haze proteins’ of wine – a summary of properties, factors affecting their accumulation in grapes, and the amount of bentonite required for their removal from wine. Rantz, J.M., ed. Proceedings of the ASEV 50th Anniversary Annual Meeting; June 19-23, 2000; Davis, California. American Society for Enology and Viticulture; 2001: 149-154.

Appendix 5. Institute Committees

Staff member Management Research Industry Communication Analytical Quality Information Biosafety Occupational Super- Staff Advisory Steering Services Steering Service System Technology Health and annuation Code Steering Steering Review Safety Negotiation

Peter HøjCCXXXX X Eveline Bartowsky XX X X Sally-Jean Bell X X X X Rae Blair X C Shauna Brown X Don Buick X X X X Dimitra Capone X Adrian Coulter X X X X Catherine Daniel X Miguel de Barros Lopes X X X Jeff Eglinton X C C Leigh Francis X X X X Mark Gishen X X X C C Peter Godden X X C X X X Jeremy Hack X Yoji Hayasaka X X X Paul Henschke X X X C Markus Herderich X X Matthew Holdstock X John Hughes XX Hans Muhlack X XX Ingrid Oats X Ken Pocock XC Alan Pollnitz X Greg Ruediger X Mark Sefton X X X Creina Stockley X X Elizabeth Waters X X C = denotes holder of Chair

63 The staff of The Australian Wine Research Institute

Acknowledgement Compiled and edited by Professor Peter Høj and Rae Blair Art direction, design and typography by Geoffrey Reed Communications Photography by Robert Geh Vintage photographs courtesy of The State Library of South Australia Printing by Five Star Press

64

The Australian Wine Research Institute