2003

h utainWn eerhInstitute Research AustralianWine The nulReport 2003 Annual 2 DIRECTOR’S REPORT Council Members The Company

Mr R.E. Day, BAgSc, BAppSc Chairman The Australian Wine Research Institute was incorporated under the South Australian Elected a member under Clause 6(e) of Companies Act on 27 April 1955. It is a company limited by guarantee, it does not the Articles of Association have a share capital and it has been permitted, under licence, to omit the word ‘limited’ from its registered name. The Constitution of The Australian Wine Research Mr J.F. Brayne, BAppSc(Oen) Institute sets out in broad terms the aims of the Institute and the Report of the Elected a member under Clause 6(e) of Committee of Review for the Institute published in March 1977 identified the the Articles of Association following specific aims:

Mr P.J. Dawson, BSc, BAppSc(Wine Science) 1. To carry out applied research in the field of oenology. Elected a member under Clause 6(e) of 2. To service the extension needs of the winemakers of Australia. the Articles of Association 3. To be involved in the teaching of oenology at both undergraduate and postgraduate levels. Mr P.F. Hayes, BSc, BAppSc, MSc, DipEd 4. To assume responsibility for the co-ordination of oenological activities, and the Elected a member under Clause 6(e) of collection, collation and dissemination of information on oenological and the Articles of Association viticultural research to the benefit of the Australian wine industry.

Professor P.B. Høj, MSc, PhD The Institute’s laboratories and offices are located within an internationally renown Ex officio under Clause 6(d) of the research cluster on the Waite Precinct at Urrbrae in the Adelaide foothills, on land Articles of Association as Director of leased from the University. The original lease is for a term of 99 years, with a right of the Institute renewal clause for a further 99 years. The Institute formally affiliated with The University of Adelaide in 1990. The first buildings were erected and opened in 1957 Mr T.W.B. James, AssDip(WineProd) and alterations and extensions were completed in 1976. The buildings have been Elected a member under Clause 6(e) of extensively modified and refurbished since that time with major extensions being the Articles of Association undertaken in 1994 and 1999, and further expansion is planned.

Mr G.R. Linton, BAppSc(AppChem), The Institute is clustered with the following research and teaching organisations: GradDip(SysAnal) Australian Wheat Management, BiometricsSA, three different Cooperative Research Elected a member under Clause 6(e) of Centres (CRC), including the CRC for Viticulture, three divisions of CSIRO, the Articles of Association Department of Water, Land and Biodiversity Conservation, Primary Industries and Resources South Australia (PIRSA), Provisor Pty Ltd, South Australian Research and Professor G.R. Scollary, MSc, PhD, BEd, Development Institute (SARDI) and The University of Adelaide’s Faculty of Sciences BAppSc(Wine Science), FRACI (which includes the Schools of Agriculture and Wine and Earth and Environmental Charles Sturt University Representative under Sciences). Currently under construction to expand the research cluster is a new Clause 6(c) of the Articles of Association building to accommodate the Australian Centre for Plant Functional Genomics (APFG); Australian Genome Research Facility (AGFR); and Australian Grain Professor S.D. Tyerman, BSc(Hons), PhD Technologies (AGT). 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

Postal Address:

PO Box 197, Glen Osmond, SA 5064

Telephone (08) 83 03 66 00 Fax (08) 83 03 66 01 Internet: www.awri.com.au

ABN: 83 007 558 296

Acknowledgement

Compiled and edited by Professor Peter Høj, Professor Sakkie Pretorius and Rae Blair Design by Geoffrey Reed Communications Photographs by Jacqui Way 49th Annual Report

Presented to The Australian wine industry

30 June 2003

Two Chairman’s report

Three Council notes

Four Director’s report

Six Staff

Seven Highlights of the year

Nine Staff activities

Eleven Research Teams’ reports

Thirty-five Industry Services Teams’ reports

Fifty-five Abridged financial report

Fifty-seven Appendices 1–5

2 Chairman's report

The year that has just passed seems to have done recognized, factored in and appreciated as an producing very valuable spin-offs which could so very quickly, but there is time to reflect on a investment in greater research productivity. never be planned, budgeted or anticipated. number of events with significant portent for the future of The Australian Wine Research Institute. During the year, the AWRI made a submission to In recent months, the process of examining the Winemakers’ Federation of Australia’s R&D existing resources in conjunction with our Provisor Last year, when our Director Professor Peter Høj Review. The fact that the peak political body of our partners has highlighted the limitations imposed accepted the Maurice O’Shea Award for the AWRI industry places sufficient importance on R&D to on our researchers by the physical constraints of he did so on behalf of 47 years of history at the undertake a review of its direction and routes to its our existing buildings. The rapid growth in R&D forefront of wine research and on behalf of the destination, speaks volumes for the ongoing role of output from highly focussed medium term projects combined efforts of several hundred past and R&D. Equally, this review process offers a valuable being funded from a very competitive funding present research staff and office holders. No doubt opportunity to improve communication of the environment, has invariably meant that we are, in the founders of the Institute would have been benefits of R&D to the industry at large. It presents wine R&D, on ‘a beer budget.’ The rhetorical pleased to see their vision recognised. It seemed also the opportunity to drive home the point that question is: How do we identify the wine R&D that this award underlined one of the intangible while, by many measures, we may pat ourselves philanthropists who will catalyse provision of the aspects of the Institute – its broader role in the on our collective back and claim great success, our kind of Institute worthy of a $4 billion industry wine industry. That the Institute has always had a spending on R&D is neither large nor assured. and soon to be demanded by it? minor and reluctant role in providing independent and objective commentary, when needed, is In what will no doubt prove to be a very important Last year I mentioned the unflagging energy and obvious enough if one cares to examine wine assemblage of research work for the industry, the task orientation of the Director, Professor Peter ‘issues’ of the past. Indeed, it is hardly surprising AWRI has, during the last year, made a start on a Høj, as deserving of special mention. During the that this role should have evolved in an industry major research project into the area of Brettanomyces last year, in very difficult personal circumstances, that uses technology as an important part of its spoilage of wine. What Brettanomyces spoilage Peter has made it even more difficult to choose positioning. Our involvement in providing the seems to lack in straight out destructive capability, worthy descriptors to record his efforts. I would technical and research input to support the compared to some other forms of spoilage, it more also like to acknowledge the efforts of all staff regulatory activities of the AWBC has been than makes up for this in terms of its insidious nature. who have been so supportive both on a personal longstanding and essentially a behind-the-scenes As a spoilage at the near edge of the envelope, it and professional basis. activity. The current trend is for technical trade produces a great deal of emotive response from barriers to be invoked more frequently and with ignorance and denial to preoccupation and paranoia. Many Institute staff members again put a great more vigour and emotion than in the past and, if As a result, collecting the valid observations is deal of effort into extending the message into the this continues, the AWRI role in these areas is difficult and the science shows every sign of being industry via Roadshows, seminar and conference likely to become more important. We don’t make more and more complex as greater information is presentations and workshops. It is particularly policy at the AWRI, but the willingness and the uncovered. The successful prosecuting of this little gratifying to see staff members blossom into expertise of our staff means that we are well criminal critter will be a great challenge and will be excellent presenters, a process which is reflective placed to help those who do, with the goal of indicative of a cohesive research and production of the pride and professionalism applied to their achieving a holistic success for our industry. community working together. primary research tasks.

Early this year, we were very pleased to welcome Some years ago, the work of our researchers on Finally, I would like to thank my fellow Councillors Professor Isak (Sakkie) Pretorius to the position of the key flavour compounds of oak cooperage led for their energy and thoughtful insights in helping Director of Research. Even for an incurable to techniques being developed to analyse the to shape the strategic direction that we hope will workaholic, the exercise of reading Sakkie’s CV is a volatile phenolic constituents of wine, one of enable us to meet best the next set of challenges. very revealing one and likely to result in feelings of which (4-ethylphenol) was shown to be a key having underachieved. Sakkie, who has quickly indicator of activity of Brettanomyces. This analysis settled into life at AWRI, has strengthened an has now become one of the most frequently already vibrant team and we look forward to his demanded of our Analytical Services team. In a contributions in the future. The Institute is very broadly similar way, a secondary output from a key appreciative of the GWRDC Board’s role in research project again seems highly likely to facilitating this important capture for the provide an extremely useful tool for industry in a Australian wine industry. completely unrelated application. Demonstrations Robin Day that juice protein profiles can be successfully used Chairman One of the potentially most rewarding challenges to fingerprint different varieties have resulted in faced by the Institute in the last year has been that numerous requests from industry to conduct juice of working together with its other shareholder fingerprinting analyses to aid in issues of varietal partners in the genesis of Provisor (formerly integrity. Currently, these techniques are being NWIRC). Notwithstanding the potential challenges tuned so that they can be offered as routine Provisor so clearly can create for existing service analyses by Analytical Services. These are prime providers, the Institute was instrumental in its examples of the value of strategic research creation as it seemed to be an opportunity too good to miss for the wine industry. While the benefits of an additional entity in grape and wine research will be very clear as its operations gather momentum, the intangibles will also have significance for the way its shareholders work together in the future. Provisor will provide not only the bridging bricks and mortar between The Australian Wine Research Institute and CSIRO Plant Industry, but it will become the cultural cement between grape and wine research and strategic and programmatic research. The obligation for the AWRI, CSIRO Plant Industry, The University of Adelaide and SARDI to work more closely together will add to the critical mass of grape and wine research in Australia.

In today’s world of multiskilling, cross disciplinary co-operations and virtual research institutions, the building of these types of links are not so much surprising but necessary, but they do place additional loads on the communication and administrative processes which support the Chairman’s report science. The costs of these loads need to be

2 Robin Day - Chairman Council notes Jim Brayne Rob Walker Tim James Geoff Linton Steve Tyerman Geoff Scollary Robin Day (Chairman) Peter Hayes Peter Høj (Director) Absent: Peter Dawson

Chairman Council At the Council meeting held on 29 October The Council of the Institute met on the 2002, Mr R.E. Day was elected Chairman following dates: 23 July 2002, 24 July 2002, of Council. 29 October 2002, 25 February 2003, 5 May 2003. Executive members of Council met Members of the Executive Committee on 3 December 2002. Mr R.E. Day Professor P.B. Høj Funding Professor S.D. Tyerman The Council of the Institute acknowledges Mr T.W.B. James the continuing financial support of the Mr G.R. Linton Grape and Wine Research and Development Corporation. Deputy Members of Council Mr N.P. Blieschke Appreciation Mr L.P. Deans The Institute acknowledges the assistance Dr P.R. Dry and cooperation of the following Mr A.M. Kennedy organizations throughout the year: Mr D.J. McWilliam Mr J. Northey Australian Wine and Brandy Corporation Dr. N.S. Scott Charles Sturt University Commonwealth Scientific and Industrial Audit Sub-Committee Research Organization (CSIRO) Mr R.E. Day Cooperative Research Centre for Viticulture Mr P.J. Dawson Department of Agriculture, Fisheries and Mr T.W.B. James Forestry Australia South Australian Wine and Brandy Industry Meetings Association Inc. Ordinary General Meeting State Departments of Agriculture The 48th Ordinary (Annual) General State Government of South Australia Meeting was held on 29 October 2002. The University of Adelaide Winegrape Growers’ Council of Australia, Inc. Winemakers’ Federation of Australia Inc. Council notes 3 Director's report

Solving problems is no longer sufficient; defining of the wine. Passion-fruit character in white 1. Establish the target concentrations of and unleashing hidden potential is a new wines is contributed by 3MHA (3-mercaptohexyl 3MHA and IBMP that give rise to the competitive imperative acetate) at such low concentrations. This desired sensorial property of the wine. aroma compound is exceedingly potent, for Such research involves repeated The Australian grape and wine industry is a example, one gram of 3MHA would be, in correlations of sensorial assessment and complex mix of different sized companies principle, enough to impart a passion-fruit analytical chemistry of preferred wines. who collectively export $2.4B worth of wine character in a quantity of liquid equivalent per annum and who share a vision of what to the size of 100 Olympic-sized swimming 2. Establish the origin of the target world wide consumers expect and will pools. Similar potency can be ascribed to the compounds. Are they derived directly continue to demand. The industry invests in compounds giving rise to the ‘green’ and from grapes or formed during R&D and has reaped benefits from adopting herbaceous capsicum character in white wines, winemaking? As it turns out, IBMP is new technologies and processes to keep which can be attributed to isobutyl derived directly from grapes but 3MHA is testing and challenging the world stage. methoxypyrazines (IBMP). The current not. The latter is formed from a precursor challenge and focus for wine and grape in grapes during microbial fermentation. The changing focus of grape and wine research: researchers is to be able to accurately Researchers would thus typically have to In past decades, much grape and wine research control the composition of wine with respect identify the precursor in grapes. has focused on identifying and managing problems in the vineyard and winery. Research has played a key role in underpinning industry growth, by reducing risks such as disease in the vineyard; microbially induced faults; and suboptimal closure performance. Parallel aspects of research and development have related to reducing labour inputs through mechanisation in both vineyards and wineries to deliver quality products at competitive price points. Significant challenges still remain in dealing with issues such as smarter application of irrigation water; further reduction of unintended microbial activity; better ways to close a bottle to ensure durable capture of quality; and to reduce production costs further. Nevertheless, there is also a desire to stretch the boundaries to improve wine. Tailored production inputs can unleash hidden potential that will enhance product attributes and value.

The holy grail and its elusive nature: One of the grandest R&D aspirations of the wine industry Peter Høj accepting the 2002 McWilliam's Wines Maurice O'Shea Award is to develop objective measures for grape on behalf of The Australian Wine Research Institute and wine quality, a feat that despite some considerable progress still remains elusive. Not only is the composition of wine challenging to unravel, but changing consumer and market preferences will dictate the type of measurement to potent impact compounds such as 3MHA 3. Assuming now that the origins of the that will be important. For example, in the and IBMP through inputs across the whole target compounds are fully understood, 1960s Australian domestic production production chain from vineyard to consumer researchers will have to establish which comprised in excess of 75% fortified wine palate. The challenge can be illustrated with vineyard practices influence the abundance and it was not until the early 1970s that reference to these two compounds in the of the target and precursor compounds in Chardonnay was first introduced by Murray following hypothetical. the grapes. For example, in the case of Tyrrell to Australia. In contrast, fortified wine IBMP, it is now firmly established that now accounts for less than 5% of Australian An illustration of the approach to current research: exposure of grapes to sunlight diminishes wine production while Chardonnay comprises Assume an extensive marketing survey of the concentration of this compound. almost 20% of total wine production. Readers consumers has concluded that there is a Accordingly, if a low capsicum character is of the many informative ‘wine columns’ now niche for a white wine which has a strong required, viticultural practices would be available will also be aware that in addition to hint of tropical fruit characters (including implemented to ensure good to moderate consumer preferences changing over time, the passion-fruit) with only a moderate herbaceous bunch exposure of the grapes. A second ranking of quality attributes also varies amongst capsicum component. Assume also that level of complexity is introduced at this individuals. The research challenge is to set extensive research had shown that of the stage, namely that of vineyard variability. clear goals from these variable messages, so that 800 or so known volatile compounds in wine, Using the tools of precision agriculture, management options for the grapegrowers the two target compounds 3MHA and IBMP ongoing research has clearly established and winemakers can be tested. alone accounted for the tropical and capsicum that considerable variability with respect to characters of wine and that all consumers vine performance exists in many vineyards. The nature of the technical challenge: Wine has are equally sensitive to the two classes of An important aspect of ongoing research been drunk for millennia but it is only in the compounds (both assumptions represent a is thus to define and manage variability to last sixty years we have understood the gross oversimplification, the reality is much pick like fruit with like fruit in order to universal biochemical pathway for fermenting more complicated). In order to assist the achieve the target compositions better large quantities of sugar to alcohol. Today, Australian wine industry to make wines that with respect to key impact compounds wine researchers and innovators are grappling fit the target niche market, the following such as 3MHA and IBMP. with how to identify, measure and manipulate research activities (which are illustrative of compounds which, when present in levels as much research taking place today) could 4. Once the grapes are at the winery, choices

Director’s report low as 0.000.000.001 grams per litre, be envisaged: about processing will have to be made. 4 significantly change the sensorial attributes For example, are some of the target in thedailyapproach toimproved production. that iscatalyticand encouragesfurthergains such innovative/questioningactivity –afact by individualsthathaveseenthesuccess from activity. TheAustralianindustryischaracterised of thisinformationintoacommercial that willjoinintheprocess oftranslatingall under appliedconditions;andanindustry knowledge;itstestingandexamination new rely onbasicstrategicresearch tounleash outcomes industry. Asalways,high impact specifiedoutcomes achieve as desirable by that isabletoapplyrelevant informationto sophisticated andwell-educatedworkforce institutions. Itrelies onanincreasingly collaboration withininstitutionsandbetween disciplinary andwillrequire enhanced The approach required isclearlymulti- advantage fortheAustralianindustry. competitive market continues toprovide expectsindustry of its the outcome.Italsoreminds usofwhat the investment andseveralvintagestodeliver work. Itmaytakemillionsofdollars some goodluckforthewholeprocess to growers andwinemakers,gooddesign, and laboratory work,closeinteractionswith will involvemanyresearch gr us remember thatthetaskoutlined above and scienceformanydecadestocome.Let wonderful combinationofart,experience winemaking will,therefore, remain a that thisisaverychallengingtaskand Sounds simpleenough,butneedlesstosay 5. 3MHA andIBMPcanthusbeset. of release, thetargetconcentrationsof is likelytobeconsumedwithinoneyear knowledge thatmore than90%of wine compounds the concentrationoftarget which at speed relative the here istounderstand the wine’s span of lifeinbottle.Thekey achieved through arelatively shorttime concentration of3MHAandIBMPisonly differ andthattheoptimalrelative two the of the stability assume that,inthishypotheticalexample, to differing stabilityinwine.Itwouldbesafe are crucialasourtargetcompoundshave sold andconsumed?Theseconsiderations expect the product to hit themarket,be choose?Whenwouldwe to What closure aroma impartingcompoundishit. whether thetargetforpassion-fruit is, therefore, amajordeterminantof strains andthecorrect choiceofyeaststrain derived precursor isverydependentonyeast efficiency ofitsformationfrom thegrape- not so,howeverinthecaseof3MHA, In thecaseofIBMPthisisprobably concentration ofthetargetcompounds? – willsomeyeastinfluencethefinal plethora ofyeastsavailabletowinemakers to thejuicecomponent.Further, ofthe proportion oftheskincomponentrelative skin contactwillincrease therelative contact timeisthenrequired aslonger in thejuice?Judiciouschoiceofjuice/skin components present intheskinandothers Now tobottlingofthefinishedproduct. change. Combinedwiththe R&D: anoutcomethat target compounds oups, fieldand wine industry’s bottomline. and itsdirect importancefortheAustralian places onthepastandpresent staff activities constitutes arecognition ofthevalueindustry Award by awarding theprestigious > events of2002/2003: credentials todoso;abeliefunderscored by this capacityandbelievewehavethe looking forward toservingtheindustryin position totakeonthischallenge.We are cross-disciplinary research, isinaprime achievement andstrengthened focuson and theInstitute,withitsrichhistoryof more challengingthanpastresearch efforts phase ofR&Dactivityislikelytobeeven hidden potentialinourgrapematerial.This but also,asillustratedabove,tohelpunleash wine industrytoaddress andavoidproblems for. ThisisnotonlytohelptheAustralian Institute, acontributionweare mostgrateful 38% ofitstotalR&Dbudgettothe a competitivebasis,awards approximately Research andDevelopmentCorporation, on not surprisingthattheGrapeandWine industry withstrategicR&Danditis,therefore, proud 48yearhistoryofservingthewine Research Institutehasa The AustralianWine FederationofAustralia. by theWinemakers’ through acollectivelevyincrease assuggested that investmentinarelatively fastmanner hoped thatthewineindustrywillenhance competitive globalindustryanditistobe maintain thepositionasaleaderinsuch sure, more R&Dinvestmentisneededto impressive achievementbutonething isfor past twodecades.Thisisindeedan the over growth inAustralianwineexports the industry afloatwhilewatching required tokeepatraditional There, approximately $2Bofsubsidiesare Europe. in just askproducers andtax-payers industry, notabadreturn onin return ofabout$150-160Mp.a.across the labour inputsetc.Thistranslatesintoa genuine improvement inquality, reduced produced byatleast10centsthrough wine of bottle per line improved the bottom years orsowouldhave 30 past the over Report, mostpeoplewouldagree thatR&D A certainly yes.Asoutlinedinaprevious sentiment is almost contributions, thegeneral prepared seem toincrease theirlevy As theAustralianwinemakersrepeatedly – istheindustryreceiving valueformoney? industry contributes Through theGWRDC, co-investment. on industryandCommonwealthGovernment Corporation (GWRDC),apartnershipbased Research andDevelopment and Wine wine R&DinAustraliaisfinancedbytheGrape to reach thesegoals.Themajorityofgrapeand Finally aword ontheexpenditure necessary and observations. informed andprioritisedbypracticalgoals wine industryisthatthebasicresearch is Au the of A secondcharacteristic Industry itselfhasrecognised ourcontribution to theInstituteinAugust2002.This about $8Mp.a.toR&D Maurice O’Shea non-innovative vestment - stralian 200-fold nnual Peter Managing Director of Research. Pretorius totheInstituteinrole asDirector our capacitybyattractingProfessor Isak countries. In2003,wefurtherstrengthened competitive efforts from otherwineproducing much harder pressed tocountertheinevitable and withoutwhichourindustrywillbe would takedecadestoassemblefrom scratch This represents atrulynationalcapacitythat committed toservingitsindustrystakeholders. professional staff thatistotally highly > astringency, understanding offactorsaffecting as wellnew pigments red wine stable of characterisation scalping’ important insightsinrespect of inthisreport. Theseincludethe elsewhere commercial relevancedirect are reported > Yet again,many wine microbiologists. recognised asoneoftheworld’s leading BiotechnologyinStellenboschandis Wine Foundation Director oftheInstitutefor doubled its turn-over since1997. doubled itsturn-over Analytical Servicewhichhasalmost our having madeatremendous contribution to to MrDonBuickwhoelectedretire after Manager withLallemandAustraliaandalso has takenuparole asNationalTechnical who, aftermore thanasevenyears service special thanksgoestoDrPeterCostello investors andInstitutestaff. Thisyear, a strong partnershipwithindustry, R&D Institute R&Dcontinuetodependona Past, present andfuture achievementsin for technologydiffusion toindustry. annum andhenceconstitutesamajorportal conducts inexcessof commercial AnalyticalServicewhich now > Continuedandincreasing supportforour spoilage ofwineby and developmentcanreduce unwanted yeast strains, The Institutehasauniquecollectionof Høj by closures, discoveryand commercial applicationof early signsthatInstitutesupport Professor IsakPretorius flavour compounds research highlights

Brettanomyces 54,000

analyses per ‘flavour , better etc. was the tannin with new 5 Director’s report 6 Staff Staff Peter BordierHøj, PaulHenschke, Anthony Research teammembers Isak StephanusPretorius, Miguel Antonio deBarrosLopes, Robert GeorgeDambergs, Elizabeth Joy Waters, Sefton, Mark Aidan Ian LeighFrancis, Gordon MichaelElsey, DipEng Yoji Hayasaka, Markus JohannesHerderich, Diego Torrea, Paul JosephChambers, Patrik RaymondJones, PaulSmith, Alexander George Kyriakos Skouroumounis, Alan Percy Pollnitz, Eveline JuttaCharlotteBartowsky, Kenneth Frank Pocock, Stéphane Vidal, Daniel Cozzolino, Dimitra LiacopoulosCapone, Kate Alexandra Lattey Kate Alexandra Tracey EllenSiebert, TangerineParker, ‘Mango’ Lorelie Flood, Jeffrey MarkEglinton, Peter James Costello, Leslie JosephJanik, Director ofResearch MSc(Agric), PhD, Managing Director UQld, Principal Research Biochemist Principal Research Chemist Principal Research Microbiologist Senior MolecularBiologist UC (SantaBarbara), Senior Research Chemist UAdel., Visiting ResearchVisiting Fellow Hertfordshire (commenced 14April2003) Research Chemist (commenced 5May2003) Research Chemist Research Chemist Flinders, Computer SystemsOfficer Research Chemist Research Chemist Aberdeen, Manager—Mass Spectrometry Facility Monash, MPharm November 2002) Postdoctoral Research Fellow(concluded22 Lyon, App. Sc.Lyon, PostdoctoralResearchVisiting Fellow Navarra UAdel., Senior Chemist USthAust., Chemist Chemist/Sensory Analyst Senior ComputerSystemsOfficer Microbiologist Chemist (commenced 20November2002) Research Assistant PhD Technical Research Officer MAppSc UAdel., PhD Senior Research Chemist Senior Research Chemist Research Microbiologist (commenced 30September2002), PhD, GradDipOenol. Vic. Col.Pharm., USthAust., USthAust., BSc, PhD, BFTech Chemist UJoseph FourierGrenoble, Research Chemist Eng Dip.(Biochem), Microbiologist AgricEng (commenced 6January2003), BSc(Hons) MSc, PhD, MBiochem, BSc(Hons), PhD BSc(Hons), PhD AssDipIndChem BSc BSc, PhD BSc (Hons),MSc Orange FreeState , BSc, (IndChem) Xavier, BSc(Hons), PhD BSc(Hons) BSc(Hons), PhD, BAgSc, PhD BAppSc BSc(Hons), PhD BSc(Hons), PhD Universidad Publicade UAdel., BSc(Agric)(Hons), BSc, BSc(Hons) BAppSc, AssDip(Chem) Uruguay, Canterbury, PhD Monash, BSc UCopenhagen Flinders, CertIntBusMgt UAdel., UAdel., UClaude Bernard BSc BSc(Hons) UWuerzburg, UAdel., Tokyo I.T., BSc(Hons), PhD UNew England, UAdel., UAdel., UAdel., UWA, Oregon UAdel., USthAust., Nat. Instit. PhD, PhD Flinders, UNSW, Flinders, FAIFST, FAIFST, , UAdel., , PhD , PhD Simon JustinDillon, Shauna LiamBrown, Maria JolantaKwiatkowski, Wieslawa Cynkar, Gayle Ann Baldock, Gayle Ann Holger Gockowiak, Maria JosephinedeSa, Daniel, Merran Alida Jane MelissaMcCarthy Jennifer RoseBellon, Kevin HerbertPardon, Anthony JohnHeinrich, Christophe Guirado, Antonio Grimaldi, Jennifer Gardner, Kate SusanHowell, Agnieszka Janusz, Carolyn Jane Puglisi, Muhlack, Richard Anthony Oenone JeanMacintyre, Heather EuniceSmyth, Matthew CarlyleCaldersmith, Kerry Leigh Wilkinson, Melissa Fettke, Julia StephanieCrossman, Jennifer Cartwright, Rachel ChristineBrown, Nicola ReneeSleep, Sally-Jean Bell, Godden, Peter William Industry Servicesteammembers* supervised byInstitutestaff See Appendix3fordetailsofallstudents Jaromir Guzinsky, Adrian DermottCoulter, Mark Gishen, Research Assistant Technical Officer Officer/Casual Analyst (from 28April–30June2003) Research Assistant Technical Officer Postgraduate Student Laboratory Technician Laboratory Manager (until 25October2002) French PostgraduateStudent Postgraduate Student Postgraduate Student Postgraduate Student BSc(Hons) TAFE, USthAust., Postgraduate Student Postgraduate Student UAdel., UAdel., Postgraduate Student UAdel., Postgraduate Student Honours Student Honours Student AnalytChem), Postgraduate Student Flinders, Honours Student Honours Student AnalytChem), Roseworthy, UMelb., Industry Services UAdel., Oenologist GradDipOenol, Technical Assistant Winemaker/Manager Winemaker/Manager Postgraduate Student Postgraduate Student Postgraduate Student Quality LiaisonManager Honours Student BE(Chem)(Hons), MEngSc(Chem) Brunel, CertVetNurs, CertAnimHand BSc(Hons) BSc(Hons) PhD BSc BBiotech, BSc, PhD Flinders, Flinders, MSc (equiv.) BSc(Hons) BSc(Hons) BSc(Hons) UAdel., BSc(Hons) BTech(For’s &AnalytChem), BSc(Hons) BAgSc(Hons) MSc BBiotech(Hons) BSc BTech, BSc(Hons) Technical Assistant , AdCertMedLabSc BAppSc(Wine Science) BAppSc(Wine UWA, UAdel., AssDip(AppChem) BSc(Hons) BSc(Hons) MSc BBiotech(Hons) BSc BTech(For’s & BSc, BE(Chem)(Hons), Flinders, ENSBANA, BTech(For’s & BE(Chem)(Hons), UWA, Flinders, MSc Honours Student Honours Student Wroclaw, Flinders, UAdel., Nottingham, Viticulturist BSc(Hons) Flinders UAdel., UNSW, Guelph, Flinders, Florence, Gliwice, Flinders, GradDip(Wine) UAdel., UAdel., BSc(Hons) Flinders, Visiting , Part time Flinders, Flinders, Technical UAdel., SAIT, Hans EngelbertMuhlack, Administration projects. lead ortakepartinanumberofresearchTeam *Several membersoftheIndustryServices Melissa ElizabethFrancis, Ingrid Betty-MaudOats, Catherine GraceDaniel, Geoffrey DavidCowey, Heather MargaretDonnell, Rachel LeeEdwards, Raelene JoanBlair, Ella MargaretClareRobinson, Creina StandishStockley, Pauline Jorgansen, Rhonda IrenePacker, Jelena Jovanovic, Emma-Kate White, Julie McConnell, Donald RobertBuick, Analytical Service Peter CharlesHansEichinger, Matthew GrantHoldstock, Sandra MargaretLloyd-Davies, Gregory Andrew Ruediger,Gregory Andrew John BenjaminHughes, Randell Leith Taylor, Maria ConcettinaMills, Athina Massis, Danielle Kylie Leedham, Matthew James Cream, Anna Catalano, Heather MandyBrooks, Belinda Bramley, Amanda LouiseCook, David RolfeBoehm, Melb. StateCol. Library Technician RMIT, Librarian Health andRegulatoryInformationManager Company Secretary Flinders, UAdel., Chemist (commenced4November2002) Director/Conference Manager Manager/Personal Assistanttothe Communication andPublicity (commenced 2October2002) to theDirector (commenced 18March 2003) (commenced 10March 2003) Function Support Analytical Service(retiring 1July2003) (BusAdmin) Customer ServiceOfficer (commenced 2June2003) UAdel., (concluded 13September2002) Administration Analytical ServiceSupervisor– Supervisor –Laboratory GradDipOenol Trace AnalysisLaboratorySupervisor GradDipOenol Casual LaboratoryTechnician Casual LaboratoryTechnician Casual LaboratoryTechnician Casual LaboratoryTechnician Mackay Administration Support , SeniorLaboratoryTechnician Chemist Manager –AnalyticalService MBA DipAppSc, USthAust., Casual LaboratoryTechnician Administration Support Casual LaboratoryTechnician Laboratory Assistant USthAust., UAdel., UAdel., Receptionist Administration Support CertAppMgt(Marketing) BSc BSc(Hons) Accountant Library Assistant Administration Officer BSc AdvCert (LabTech) BSc(Hons) Analytical Service DipWineMrktg DipWineMrktg BA UAdel., DipLibInfo BSc(Hons) BEc BA UAdel., BAppSc BSc Secretary USthAust., AIFST, Manager– Analytical Service ANU BSc(Hons), PhD UMelb UAdel., BA, BSc(Hons) BA Flinders UAdel., Technical Officer , GradDip(Lib) UAdel Flinders, GradDip SAIT, Adel. Tafe, ., DipEd UAdel., CPA Aust., , UAdel., Chemist ., MSc AIM, Highlights of the year

The Institute was awarded the prestigious were intermediates in the formation of the The extent to which the discriminating power McWilliam’s Wines Maurice O’Shea Award in important wine aroma compound damascenone. can be extended to white wine is limited if recognition of its contribution to the success These findings have led us to re-evaluate the bentonite fining has occurred. However, as DNA of the Australian wine industry over the past mechanism by which damascenone is formed typing appears not to be practically possible 48 years. in wine and other food products and will for wine samples, the extension of our facilitate our goal of being able to improve findings might represent one of very few The Institute attracted one of the world’s very best wine microbiologists, Professor Isak wine quality by manipulation of wine avenues by which varietal authentication of Pretorius, to the position of Director of composition in the vineyard and in the winery. white wine can be executed with some degree of certainty, if necessary through a Research with effect from January 2003. This Glucosides of the open-chain forms of cis- and augurs well for the future of the Institute trans-oak lactone have been synthesised. These combination of analyses of secondary and it is a reflection of the high esteem in compounds are stable in solution at wine pH, metabolites and, of course, sensory analyses which it is held. The strategic foresight of but are converted to the corresponding for varietal typicity. the GWRDC in assisting this critical lactones at barrel-toasting temperatures. This Milligram quantities of a haze-protective factor development is acknowledged gratefully. demonstrates that such compounds can act (6xhis Hpf2p) have been overexpressed in as precursors of oak lactone during barrel yeast and purified. This has allowed us to Using organic synthesis we proved the structures of two malvidin-derived wine pigments and manufacture, but not during subsequent wine treat a wine and assess its short to medium established their colour properties. With the conservation. By separating the isomeric term stability during storage at 25˚C. After help of these synthesised reference compounds glucosides from each other we have also been three and six months’ storage, 6xhis Hpf2p we demonstrated that the pigments, which we able to resolve the nature-identical isomer of had retained full haze-protective activity, had detected previously for the first time in cis-oak lactone from its non-natural optical an important prerequisite for commercial red wine, were almost inert to SO -mediated isomer and evaluate its sensory properties. viability in relation to the prevention of 2 Previous sensory work on cis-oak lactone in protein hazes in white wine. bleaching, and that they showed superior wine has been carried out on mixtures of these colour properties and enhanced extinction Methods to measure some volatile thiols of two forms (i.e. the racemate), or even on coefficients in model wine at pH 3.6 when importance to varietal flavour have been mixtures of cis- and trans-oak lactone. The compared to grape-skin anthocyanins. developed and are now being applied to sensory detection threshold of the nature- identify commercial wine strains that release A method for the measurement of acetaldehyde identical isomer of cis-oak lactone (which is ten either high or low amounts of these important has been developed and validated. This times more potent than the trans form) in a aroma compounds. These findings will enables us to monitor acetaldehyde neutral dry white wine was 23 mg/L. This is initially be important for modifying the concentrations in various tannin-, yeast- and much lower than the value of 90 mg/L flavour of a wide range of varietal wines. wine oxidation-related research projects and previously reported for the racemic mixture. will assist in developing a more detailed A collaboration with The Australian Proteomic We have demonstrated that at wine pH, (3.0 – picture of this reactive aldehyde. Analysis Facility (APAF) in Sydney has 3.8), oak lactones exist almost entirely in the permitted the identification of proteins that With the help of rapid HPLC methods developed aroma-active lactone form at equilibrium, in are differentially expressed in wine yeast strains. by the Tannin team, more than one contrast to an earlier hypothesis. At the highest In the longer term, it is hoped that this thousand samples from research projects of pHs studied, however, the open chain form of research will identify factors that provide wine the Tannin, NIR and Microbiology teams the cis-isomer lactonised relatively slowly, yeast with their important winemaking have been analysed to establish indicating the need for suitable soaking times properties and is expected to lead to the anthocyanin, tannin and 'pigmented polymer' and low pHs of wine or aqueous extracts when generation of improved strains. concentrations in samples from viticultural preparing extracts of oak shavings for and winemaking trials. These data are vital analysis. The data show that wines treated Flavour metabolites that are produced at to improve our understanding of the with oak chips or shavings over a few days much higher concentrations in Saccharomyces correlation between grape polyphenols, and could continue to develop 'oakiness’ by species that are generally not associated the composition, colour and mouth-feel of additional formation of cis-oak lactone once with winemaking have been identified. wine. They also form the basis for our the chips or shavings are removed. These results indicate that interspecific investigations into the potential of NIRS for hybrid yeast offer an effective strategy to Sensory studies robustly established that purified the efficient measurement of polyphenols. harness diverse flavours in wine. anthocyanin glucosides and their corresponding Using solvent step-gradients with multilayer coumarates were not perceived as bitter or Winery trials made in various wine regions coil countercurrent chromatography astringent. In addition, these grape pigments with AWRI isolates of S. bayanus yeast are (MLCCC), the isolation of anthocyanins and did not influence the fullness or viscosity of confirming our laboratory trials that have coumaroylated anthocyanins from the solutions in which they were tasted. shown that these yeast can impart a greater commercially available grape marc extracts diversity of sensory characteristics than The effect of ascorbic acid addition on oxidation was optimised. In addition, MLCCC was achieved with S. cerevisiae, especially to aroma of wine continues to be a matter for debate in instrumental in enriching very polar and and palate fullness and texture. Demand for the literature. Previous laboratory scale work potentially novel pigments for structure scaled-up winery trials with these S. bayanus from the Institute (Institute publication #577) elucidation efforts, and in isolating yeast has increased to the point that we have and others indicated an enhanced depletion 'pigmented polymers' for sensory studies. engaged Lallemand to develop commercial of sulfur dioxide in wine and model solutions starter cultures. 2,3,6-Trimethylphenylbutadiene (TPB) has been to which ascorbic acid was added but formal identified as a hitherto unrecognised wine sensory assessment has rarely been conducted A random amplified polymorphic DNA (RAPD) flavour compound that is formed by hydrolysis in these experiments. Through collaboration method for determining genetic similarity of grape-derived glycoconjugates. The with a winery we have now examined the and differentiating strains of commercial threshold of TPB in a white wine was effect of ascorbic acid addition to a Riesling and wine isolates of the malolactic bacterium determined to be 40 ng/L and sensorial and a Chardonnay wine bottled on a large Oenococcus oeni has shown that: (i) all descriptors ranged from 'floral', 'geranium' scale under commercial conditions. Aroma, commercial strains tested are genetically and 'tobacco' at the lower concentrations to colour and compositional data indicate that distinct; (ii) Australian isolates from different 'pungent', 'very green', 'unpleasant', 'plastic', for four closure types examined, these two wineries are genetically distinct and at least and 'insecticide' at concentrations of 270 ng/L wines were less oxidised after storage under as genetically diverse as the commercial and above. This makes it one of the most ideal conditions for two and a half to three strains; and (iii) isolates from a given winery potent wine flavour compounds known. years when ascorbic acid was added at may be genetically diverse. Identification of TPB in wines has furthered bottling. When analysed after six months’ The adventitious growth of spoilage acetic acid our understanding of the contribution of grape storage there was little difference between bacteria in bottled red wine stored in an secondary metabolites to wine quality. the wines (Institute Annual Report 2000). upright vertical, but not horizontal position, Four isomeric potential damascenone precursors, The potential for mass spectrometry of red and has led us to hypothesise that the upright have been synthesised. The availability of these white juice proteins to differentiate varieties storage position created a heterogeneous authentic samples allowed us to prove that they of Vitis vinifera has been further confirmed. environment that allowed the growth of a Highlights of the year 7 bacterial film in only those bottles sealed Work is continuing with further samples positive, with no identifiable areas of concern. with cork closures with substantial permeability remaining in frozen storage for later analysis The results are consistent with increases in to oxygen. Such a heterogeneous environment to investigate any long-term stability effects. revenue in the past financial year. A large would likely not exist in horizontally stored proportion of the Analytical Service activities The Institute has been involved in the testing and bottles since the larger volume of wine assisting in the development of an automated are now associated with customised non- adjacent to the cork would strongly compete grape sample preparation system ‘Bioprep5 routine projects for both Australian and with the bacteria for the oxygen as it diffuses Robotics’, which has been released commercially overseas companies. through the cork closure. The direct by a local Adelaide company. The results from Eleven thousand copies of the Institute’s measurement of oxygen content of spoiled samples prepared with the automated annual publication, Agrochemicals registered and non-spoiled wines support this hypothesis system compared well to those obtained by for use in Australian Viticulture 2003/2004 which has been published in an international the manual method in terms of accuracy and were produced in response to an increase in scientific journal (Institute publication #718), precision. The unit appears to offer potential demand. The reduction in the number of but still requires further rigorous confirmation. in speeding up the preparation of red grape agrochemical related enquiries in comparison The Industry Services’ team responded to 1,676 homogenates for scanning by NIR (up to to previous years illustrates the effectiveness enquiries during the year, and analysed 2,231 112 samples per 8 hour day) when configured of the Institute’s role in compiling and samples (an 81% increase in sample analysis to act as an automated homogeniser. distributing this publication. over the previous year). An evaluation of the feasibility of NIR A booklet entitled The A-Z of information on A record number of Roadshows were conducted, spectroscopy for detection of Botrytis and wine and health was developed and with ten seminar days and five workshops powdery mildew in grapes is continuing in published. The Commonwealth Department being held in four states. collaboration with the Fungal Pathology of Health and Aged Care favourably Laboratory of The Adelaide University and reviewed the booklet and contributed The flow of new information from the Institute's closure trial continued, with three publications CSIRO Plant Industry in Canberra. Distinct towards its publication and printing costs. during the year (two in Technical Review and spectral changes were observed in fruit that As of 30 June 2003, approximately 48,500 one conference paper [see Appendix 1]). had been visually classified into a number of booklets have been distributed. different levels of powdery mildew infection. Nineteen interviews were conducted with Staff of the John Fornachon Memorial Library media and trial results were used or referred Principal component analysis of the spectra responded to 3,608 requests for information to in a number of other articles published was used to detect and correctly classify, during 2002/2003. both in Australia and overseas. powdery mildew infection down to at least a 10% infection level. Similar work is also in Over 5,000 new records were added to the The Institute's Dekkera/Brettanomyces control progress with Botrytis-infected fruit. The web-accessible database of the Library seminar was presented on 19 occasions implication of this work is that it may be (available only to Australian winemakers and during the year to approximately 530 possible to discriminate fungal infected fruit grapegrowers) during the year, making a winemakers, and has been presented to at the weighbridge to provide a 'go/no-go' total of over 25,000 records available for approximately 1,550 winemakers since 1999. test to highlight suspect fruit for further searching, 24 hours per day, 7 days per week. An ongoing targeted survey of the composition detailed analysis to determine suitability Six issues of Technical Review published during of Cabernet Sauvignon wines from five for winemaking. the 2002/2003 financial year were Australian wine regions indicates an Stable isotope dilution analysis methods for 33 consolidated onto a searchable CD ROM apparent fall in the concentration of key and distributed to Australian winemakers Dekkera/Brettanomyces yeast derived important fermentation esters, acids and alcohols using their deuterium labelled and grapegrowers. The new website of The spoilage compounds in wines from the Australian Wine Research Institute was 2001 vintage, compared to the previous five analogues as internal standards have been fully validated. The methods have facilitated launched to industry. The new website is vintages. The basis and durability of this easier to navigate and faster to download pleasing trend is to be investigated further. accurate and rapid measurement of the composition of wines and model media and contains additional useful information. in conjunction with A trial conducted ferments and are are now being applied to General highlights grapegrower and winemaker associations a range of cross-disciplinary projects at the and wine companies into the effects of Institute including that of flavour scalping The Institute published 41 papers on Institute bushfire smoke on grapes and wines, by closures. activities in refereed and non-refereed elucidated an understanding of the nature publications. Stable isotope dilution analysis methods for wine of the problem and possible taint- Institute staff gave 174 oral presentations, minimisation strategies. taint compounds derived from petroleum products, including toluene, styrene, C2 alkyl conducted thirteen workshops and Efforts to develop a practical NIR method for benzenes (e.g. xylenes), C3 alkyl benzenes presented 20 posters. determining colour, total soluble solids (TSS) (e.g. trimethylbenzenes), C4 alkyl benzenes Institute staff presented 48 lectures and and pH in red grapes continue and have (e.g. tetramethylbenzenes), naphthalene, coordinated a six week subject to been outlined in an industry publication methylnaphthalenes, dimethylnaphthalenes undergraduate students. (Institute publication #730). During the 2003 and trimethylnaphthalenes have been fully Institute staff supervised/co-supervised 31 vintage, the total number of samples analysed validated. The methods enable us to more postgraduate students. by NIRS represented approximately 300,000 readily assess wines for possible contamination. tonnes of fruit, in excess of 20% of the total Institute staff recorded and responded to The Viticulturist responded to 415 enquiries, national harvest. Significant benefits have also 6,001 requests for information during the participated in nine Institute Roadshows and been demonstrated from the use of NIR in 2002/2003 year or, to put the statistics into five Research to PracticeTM workshops across the research environment. Using NIR to perspective, 24 people contacted the three states. predict red grape quality parameters (colour, Institute seeking information on every TSS and pH), researchers at CSIRO (Land & The Analytical Service laboratory underwent a working day of the year. This figure does Water's Precision Agriculture group) and The technical assessment by NATA to the new not include request for work through the University of Adelaide (Roseworthy campus) ISO/IEC 17025 standard. Two new test Analytical Service which conducted 54,000 have been able to significantly reduce the cost methods; namely an enzymatic acetic acid individual analyses during 2002/2003. of analysis and the necessity for wet chemistry assay and the HPLC multiresidue assay were Readers are strongly encouraged to read the in relation to their research projects. accredited and two additional staff members report in detail rather than relying on the have become NATA signatories. Preliminary results have indicated that neither points above for information. freezing of red grape samples prior to A new Analytical Service fee schedule has been homogenisation, nor the type of printed as a result of a review of the service homogeniser had any significant effect on delivery charges. In addition, an extensive the accuracy of laboratory determinations of market research report was commissioned colour and TSS. However, as one might to establish how we were meeting our client

Highlights of the year expect, there was an effect of freezing on pH. expectations. This report has been extremely 8 at TheUniversityofAdelaide. IndustryChairofOenology Australian Wine Professor Højisthecurrent holderofthe and theManagementCommittee. Agriculture andWine’s AdvisoryCommittee The UniversityofAdelaide’s Schoolof He isalsotheInstitute’s representative on Peter Højisamemberofthefollowing: Appendices are describedbelow. Activities inadditiontothosethe Appendices 1–4oftheAnnualReport. published istabulatedandcanbefoundin students supervised,andthepapers academic lectures delivered, graduate papers giventooutsideorganizations, Information onseminars,talksandposter support ofthe Australian wineindustry. performs alargenumberofexternalactivitiesin theInstitute projects describedinthisreport, In additiontoundertakingresearchandother research agencies) between Universitiesandpublicly-funded Review Committee(intocollaboration (Chair) Melbourne) Conference (24-29July2004, Twelfth IndustryTechnical AustralianWine Conference PlanningCommitteeof the des SciencesdelaVigne etduVin Editorial Board ofthe Research of GrapeandWine Publishing, publisherof Committee ofManagement,Viticultural Waite CampusManagementCommittee Horticultural SocietyofSouthAustralia) Committee(RoyalAgriculturaland Wine Viticulture II(CRCVII)Board Cooperative Research Centre for Provisor PtyLtdBoard Federation ofAustralia) Technical Committee(Winemakers’ Committee (AWBC) Compliance andTechnical Advisory Premier’s Council(SouthAustralia) Wine (South Australia) Premier’s ScienceandResearch Council Innovation Council Prime Minister’s Science,Engineeringand Journal International Australian Journal project 1.3oftheCRCVII. Sciences Wine of the Mark Sefton the CRCVII. Elizabeth Waters University ofAdelaide’s Children's Services. She isalsoaBoard MemberofThe Member, Waite CampusExecutive Committee CongressInternational andHealth Chile2002Wine Vinsalud Member, ScientificAdvisoryBoard, Group oftheOffice etduVin delaVigne Expert Vice-President, NutritionandWine etduVin de laVigne Member, AustraliandelegationtotheOffice Committee) (Sub-committee ofSAWBIA’s Environment Member, Eco-efficiency Working Group National Environment Committee Member, AWBC/WFA Industry Wine Review Committee Member, AWBC Legislation Technical Liaison) Technical AdvisoryCommittee(as Creina Stockley Technical Conference. Industry Manager oftheAustralianWine and istheTreasurer andConference Planning Committeeforthe12thAWITC Rae Blair Public Officer ofProvisor PtyLtd. Inc. HeisalsotheCompanySecretary and IndustryTechnicalAustralian Wine Conference Hans Muhlack Professor oftheUniversityStellenbosch. Planning Committee.HeisalsoanAffiliate Conference, andmemberoftheConference IndustryTechnicalAustralian Wine Program Sub-CommitteeoftheTwelfth Yeast. Microbiology, FEMSYeast Research Journal ofEnologyandViticulture, Annalsof Board ofthefollowingjournals: Sakkie Pretorius International JournalofVine and He isalsotheCo-Chairof is amemberoftheConference is ontheeditorialreview board is thePublicOfficer ofthe is aMember, Industry Wine is amemberoftheEditorial and istheproject leaderof is ManagerofProgram 1of Rachel Edwards, Rhonda Packer,Rachel Edwards, Emma-KateWhiteandJulieMcConnell American and Staff activitiesStaff Analysis Group Committee. Matthew Holdstock the fieldofchemicalanalysisfoodandwine. National AssociationofTesting Authoritiesin Don Buick group knownastheLegalMetrology Group. Australia’s (WGCA)recently formedworking (WFA) Growers' Councilof andWinegrape FederationofAustralia on theWinemakers' CRCVII, andistheInstitute'srepresentative Mark Gishen Conference. IndustryTechnicalAustralian Wine workshops tobeheldattheTwelfth Coordinator ofaprogram of~70 Sub-Committees andistheWorkshop AWITC Conference PlanningandProgram Peter Godden Health andSafetyCommittee. Sciences (Waite Campus)Occupational Eveline Bartowsky (Chair 2003/2004). (27-29November2002) Melbourne on Yeast: Products andDiscovery, a memberofthe2ndAustralianConference and MitteilungenKlosterneuburg. African JournalofEnology Research;South Journal ofGrapeandWine Board ofthefollowingjournals: Paul Henschke and FoodChemistry. Advisory Board ofthe Adelaide andheisamemberofthe Associate Professor atTheUniversityof 'Tannin project', oftheCRCVII,Affiliate Markus Herderich and Agriculture. of the Leigh Francis Journal oftheScienceFood is avoluntaryassessorforthe is leaderofproject 1.4ofthe is anEditorialBoard member is amemberofthe12th serves ontheEditorialReview is LeaderofProject 1.2,the

served ontheFacultyof serves ontheInterwinery Journal ofAgricultural

and Viticulture Australian He was 9 Staff activities 10 Visitors to the Institute Visitors to the Institute Weckert, McQuinn, Hutch Rank, September 2002) students, FlindersUniversity(27 Dr MichaelPerkins (24 September2002) Richard Gibson, (24 September2002) Paul Baggio, University ofAdelaide(13September2002) Paul Duldig, Cornish, James McWha, Improvement Association(19August2002) 35 members Chris Proud, Australia MP Adviser, Office ofSimonCrean, Martin Ferguson), Murray RadcliffeandDeniseSpinks Shadow MinisterforRegionalDevelopment, member Council ofSA, Resources, Murray DarlingDivision,DeptofWater Barbara Hardy, (12 December2002) Cornelis Versteeg, Dr DrKamaljitSingh Vikhu, Dr RaymondMawson, Services (5November2002 Programs Branch,DepartmentofHuman Keith Evans, 24 April2003) (Dr Brendan Nelson)(1November2002and Minister forEducationScienceandTraining Catherine Murphy, Foreign Affairs andTrade (22October2002) Administration andBusinessRelations, Portugal and Dr Janet Gardiner, Sales Consultant,DuPont(16October2002) Environment Minister, Mike Young, Hon JohnHillMP, (29 April2003) Bryn Haugaard, CRC forBioproducts (28April2003) Jane Evans, George Willcox, Trade (2April2003) Department ofBusinessManagementand Michael Shillabeer, Jodie Goode, Australia (20March 2003) Ian Sutton, University ofNewEngland(12March 2003) Biomedical andMolecularSciences Professor Ken Watson, February 2003) George Bowyer George Krackljack, Science Council(10February2003) Steve Morton, Reference Laboratory(7February2003) Dr LawrieBesley, Norm Cook, Hill Vineyard (30December2002) Mario Marson, (16 December2002) Hatton MP and ShadowAssistantTreasurer, – Tasmania, Deputy Vice-Chancellor, Research, District Manager, onOeih MartinFerguson John Olenich, Development Manager, – NSW(2May2003) Winemakers’ Federationof Winemakers’ Technology Transfer Manager, Simon Divecha, Director ofFinance,The Dupont (13January2003) Director, DrugStrategyand E&J Gallo(19July2002) Managing Director, CSIRO, of theClare Valley Vine Australian Winemakers, Australian Winemakers, Executive Officer, Premier’s David Haynes, Grapegrower Heathcote/Jasper AO and Vice Chancellor,Vice and Austwine ExportsPtyLtd Vaslin Bucher(24April2003) Scorpex Wine Services Scorpex Wine Senator Penny Won National Analytical Minister fortheEnvironment, KelvinMP Thompson Food ScienceAustralia Australian Ambassadorto Chief ofStaff, Office ofthe and 10medicinalchemistry ARRM (20February2003) Alex Gordon, Greg Byrne, Peter Hoey, School ofBiological, Bill Hardy David CoxMP CEO Conservation David Richards, Executive Officer SA Director, BioStart (26 Edwina Environment David Dick Adams (Staff of g andstaff Michael Nick , Shadow , SAMP MP, Keita Hori Meiji University, Japan (6September2002) Professor Takesako Japan SpA, Italy(24September2002) Manager, OenologicalProducts), Esseco Sadahiro Katunuma Jozo; Kishidaira, Nasazuni Dobashi, Yamagani Naoshi, Products) and Sam Benelli Italy Lazlo Romsics, Hungary Ampeooeniki, Greece (6February2003) Marinos,Dr Vassilios Greece Weinsberg, Germany(25July2002) fur Wein-und ObstbauWeinsberg Dr OliverSchmidt, Germany December 2003) France(12-13 etduVin, Vigne Dr Jean-ClaudeRuf, Francois Davaux, April 2003) Patricia Villavicencio, Equipe Polyphénols,INRA(10March 2003) Recherche Sciencespourl'Oenologie: Dr Vassela Atanasova, Equipe Polyphénols,INRA(9October2002) Recherche Sciencespourl'Oenologie: Cheynier,Dr Veronique France Maule Region,Chile(8July2003) Alejandro Soto, O’Higgins RegionalBranch;TodoChile, Veronica Godoy, Danilo SturizaJordan, Chile Denmark (29April2003) Laura West, (27 February2003) Ole Vestergaard, Denmark International June 2003) Willy Billiard, Australian ChemicalInstitute.(2June2003) Pidgeon, (8 May2003) Yoshiyuki Tateishi, February 2003) Hayakawa, Kenzo Akazawa, David Edmonds, Forestry Australia(16May2003) Department ofAgriculture, Fisheriesand John Power, Development Corporation(8May2003) Roger Hoare, and SA BrandPresident, Royal and Takeda Winery; Hayakawa Co.Japan,(18 (Sales Manager, Oenological De DanskeSpritfabrikker, Assistant Manager, Policy, Wine Grape and Wine Research and Grape andWine freelance Qld.(6 journalist, Kazuhiro Yamada, Hungary (10June2003) Ken Inose, Regional Director CORFO Dr GiulianoBoni Yamato Budoshu; National President; Investment Specialist, Aahus University, Denmark ITV, France(24April2003) Yumego BudoshuKenkyusho; Kirin Brewery CoLtd,Japan Kozo Tuchiya, Lehr- undVersuchsnstalt Chuo Budoshu; and fivestudentsfrom Office dela International Chemist-Oenologist, ADEPTA, France(24 Director Zonal;and Unité Mixtede Unité Mixtede Okutone Wine KK; Okutone Wine Hisayoshi Tsuiki, Kizan Yoshu; Hojo Wine; (Sales Support Noriko Mashami Ken (26 March 2003) E&JGallo,USA Winemaking; International Relations; Growing; Tom Smith, Terry Lee; Dr Mary Wagner, Foundation, USA(7November2002) Patrick Gleeson, October 2002) Leslie Norris, (11 October2002) Dr JeffreyMcCord, Dr NancyIrelan, USA (19 September2002) Oxon,UK Development, CABInternational, Peter Scott, (20 August2002) Sam Harrop, United Kingdom Charoen Charoenchai A group of30winemakers,organisedby Thailand Azorean University, Portugal(24March 2003) Lima, Associate ProfessorMaria-Theresa Ribeirode Portugal (26 February2003) Gerard Besamusca, University, NewZealand(25November2002) Chemistry &FoodScience,Auckland Paul Kilmartin, New Zealand Agricultural SciencesDepartmentof Greg Coleman, Chris Proud, Director Programme Marks andSpencer, London,UK FlavorSense, USA(11 Lecturer inAnalytical American Vineyard E&J Gallo(August2002) Chief Technology Officer; StaVin Inc,USA StaVin AgConsult, NewZealand , Thailand(17March 2003) Vice PresidentVice –Wine Director – Director –Grower Dr Dr Research Teams’ report

Investigations into the relationship between Dekkera/Brettanomyces yeast and red wine in Australia

Staff: Peter Godden, Professor Peter Høj, Professor Sakkie Pretorius, Jane McCarthy, Jennifer Bellon, Dr Paul Henschke, Dr Peter Costello, Dr Miguel de Barros Lopes, Dr Leigh Francis, Kate Lattey, Dr Mark Sefton, Dimitra Capone, Dr Alan Pollnitz, Mark Gishen, Adrian Coulter, Geoff Cowey, Ella Robinson, Greg Ruediger

Preliminary investigations have been Peter Godden & Mark Gishen conducted under a number of Institute projects during the current reporting period, as a prelude to a dedicated project that will be funded from the 2003/2004 financial year onwards. Approximately 60 isolates of Dekkera/Brettanomyces yeast have been obtained from various Australian wines by the Industry Services team. These isolates have been confirmed as Dekkera/Brettanomyces isovaleric acid is reported in the literature 1 first contacted Industry Services by Dr Miguel de Barros Lopes and Jenny to be an important spoilage compound regarding the same issue early in 2000. A Bellon of the Institute’s Molecular Biology formed by Dekkera/Brettanomyces yeast, seminar relating to Dekkera/Brettanomyces team, using Polymerase Chain Reaction (PCR) initial analytical results obtained using this control has been presented to approximately analysis. Initial amplified fragment length analytical method indicate no correlation 1500 winemakers in a number of forums, polymorphism (AFLP) analysis indicates that between the concentrations of 4-ethylphenol particularly Institute Roadshows, over a although approximately 50% of the isolates and isovaleric and 2-methyl butyric acids period of approximately four years. are genetically very similar, a considerable albeit in a small number of wines (Table 1). Additionally, the Trouble free winemaking degree of genetic diversity exists between However, it is possible that synergistic workshop conducted by the Institute, has the isolates. It is intended that the most sensory effects exist between these a strong emphasis on the control of genetically diverse isolates will subsequently compounds, which may result, for instance, microbiological instabilities including be used in small-scale fermentation in the presence of isovaleric acid Dekkera/Brettanomyces, and has been experiments. This aspect of the project will enhancing the apparent intensity of other presented on a total of 15 occasions since seek to investigate any differences between Dekkera/Brettanomyces-derived taints in it was first developed in 1999. strains in their propensity to form 4- wine. It is considered of the utmost ethylphenol, 4-ethylguaiacol and another importance that such sensory relationships The two wineries for which data are presented reported important spoilage compound, are elucidated and, therefore, sensory in Table 1 are noteworthy for the magnitude isovaleric (3-methyl butyric) acid, when investigations will be a major part of the of the reduction in 4-ethylphenol varying concentrations of precursor ongoing Dekkera/Brettanomyces project. concentrations that they have been able to achieve. However, an ongoing survey being Table 1. 4-Ethylphenol and 3-methyl butyric acid concentrations (µg/L) of Cabernet Sauvignon wines produced by conducted by Industry Services provides two Australian wineries 1996 – 2001 some evidence that a reduction in median 4-ethylphenol concentration occurred in Winery A Winery B Cabernet Sauvignon and Cabernet Sauvignon/Merlot wines produced in five Vintage 4-ethylphenol 3-methyl 4-ethylphenol 3-methyl regions during the 2001 vintage, compared µg/L butyric acid µg/L butyric acid with the previous five vintages. Wines from (isovaleric acid) (isovaleric acid) the regions Barossa Valley, Coonawarra, µg/L µg/L Hunter Valley, Margaret River and Yarra Valley have been surveyed in a 1996 NA NA 1500 1654 representative manner. One hundred and 1997 1420 973 2450 1095 seventy-seven wines from the vintages 1998 1730 1231 2130 1274 1996 to 2000, and 51 wines from the 2001 1999 393 1383 2180 1469 vintage have been analysed. There were 2000 603 1521 563 1489 no significant differences in the median or 2001 32 1484 333 1311 mean (data not shown) concentrations of 4-ethylphenol for the vintages 1996 to 2000. However, the mean concentration of 4- compounds and nutrients such as residual The data in Table 1 demonstrate that it is ethylphenol in wines from the 2001 vintage, sugars and nitrogen, are present. The possible for wineries to achieve substantial compared to the mean of the pre-2001 wines, effect of pH and SO2 concentration on the reductions in 4-ethylphenol concentrations fell by 46% (data not shown). It is also of ability of strains to form the reported by implementing winemaking strategies interest that the distribution of 4- spoilage compounds and their correlation to control the proliferation of Dekkera/ ethylphenol concentrations appears to with the intensity of the ‘Brett’ character Brettanomyces yeast. It is pleasing to have changed in 2001 compared to the will also be investigated. Industry Services staff that, for instance, previous four years, with the lower winery A in Table 1 was one of the first to approximately 62 percentile of The development of a GC-MS method for the contact the Institute regarding Dekkera/ concentrations in 2001 falling in the range quantification of isovaleric acid (3-methyl Brettanomyces control, late in 1998. During occupied by the lower 25 percentile of butyric acid) and 2-methyl butyric acid, is 1999 and subsequently, winery A and concentrations of the previous four years. reported under Analytical method Institute staff have had many discussions on Whilst these results are encouraging, care development and evaluation. Whilst the same issue. Similarly, winery B in Table should be taken when interpreting these report Teams’ Research 11 12 Research Teams’ report compounds in wine. With theappointment compounds inwine. With focussing ontanninsandotherphenolic element mouth-feel interactions,withastrong Patrik's research willconcentrateonflavour/ funding availablethrough theCRCV. the replacement with forDrStéphaneVidal in April2003asResearch Chemistandas closely withDrPatrikJones,whocommenced present inred wine.Paulwillcooperate feel andcolourproperties ofpolyphenols to determinethemolecularbasisformouth- tannins andpigmentedpolymers,aim focus onstructure elucidationofwine May 2003asResearch Chemist.Paulwill Dr PaulSmithcommencedemploymentin winemaking andageing. conditions favouringtheirformationduring red winecolourandmouth-feel,of identification ofpolyphenolsimportantto continued withafocusondetectionand tannin andcolourspecificationhas last year, ourresearch intowine grape composition, qualityandstyle.Duringthe compounds whichimpactonwine insight intoalltransformationsofgrape properties; andtodevelopadetailed that are relevant forspecificmouth-feel a molecularlevel;toidentifycompounds understand thebasisofred winecolourat The aimofthe'Tannin project' isto France: DrVeronique Cheynier Recherche Agronomique, Montpellier in August2002).InstitutNationaldela (appointed aspostdoctoralresearch fellow David Lee,RenataRistic,DrEwaldSwinny Lambert (PhDcompletedOctober2002), in July2002),DrGrahamJones,Stephanie resignation from TheUniversityofAdelaide Patrick Iland(continuesasconsultantafter TheUniversityofAdelaide:Dr Wine, Collaborators: Pardon, MangoParker(neeRanzijn). Mariola Kwiatkowski,KateLattey, Kevin part-time Technical Officer inJuly2002), Donnell, HolgerGockowiak(appointedas Gayle Baldock,MariadeSa,Heather Dr GeorgeSkouroumounis, Yoji Hayasaka, 2002), DrPatrikJones(from April2003), (untilNovember 2003), DrStéphaneVidal August 2002),DrPaulSmith(from May Dr LizWaters, DrZhongKuiPeng(until Staff: Wine grapetanninandcolourspecification which appearslaterinthisannualreport. physiologicalcharacterisolation preliminary Brettanomyces - with *See furtherreport ontheInstitute’s work regions) willhelpaddress thisissue. the muchcooler2002vintage(inmost regions in2001.Theinclusionofdatafrom Sauvignon/Merlot winesfrom thesefive SauvignonandCabernet in Cabernet fall intheconcentrationof4-ethylphenol winemaking mightaccountfortheapparent factors duringthegrowing seasonand data, asitispossiblethatclimaticorother Brettanomyces Dr MarkusHerderich, DrLeighFrancis, School ofAgriculture and isolation fromwineanda under Dekkera/ Paul Smith,MangoParkerandHolgerGockowiak we demonstrated thepotentialofNIR thiscollaborative study,calibrations. With temperatures were usedtodevelop NIR two different yeasts,andthree fermentation additional variablestwotypesoffermenters, and Shiraz),alsoincludingas Sauvignon two grapevarieties(Cabernet wine fermentationsovertwovintages using 495 samplesfrom 32commercial-scale red team. HPLCreference datafrom atotal of fermentations incollaborationwiththeNIR phenolic compoundsinred wine spectroscopy fortherapidmeasurement of evaluate thepotentialofnearinfrared (NIR) operations further, wecontinued to To improve theefficiency ofouranalytical Microbiology teams. NIR andWine trials, andresearch projects oftheTannin, samples from viticulturalandwinemaking handle more thanathousandresearch in infrastructure enabledustoefficiently methods forHPLCanalysis,thisinvestment Together withourimproved, rapidandrobust 'proanthocyanidin' or'condensedtannin'). this report 'tannin'refers exclusivelyto 'pigmented polymers'(notethatthroughout routine analysisofanthocyanins,tanninsand for analyticalmethoddevelopmentand now three fullyoperationalsystemsavailable installed andcommissioned,wehave A newgradientHPLCsystemhasbeen Analytical methoddevelopment Roadshow seminars(seeAppendix1). emerging knowledgewithIndustryduring conferences andefficiently discussedthe scientific communityatinternational the outcomesof'Tannin project' tothe and teammemberswere invitedtopresent which appeared duringtheyear(Appendix4), progress asevidentfrom itsmanypublications m team studies. Theremodelled and essentialstartingmaterials for sensory purification ofcrucialreference compounds anthocyanins andHolgerwillhelpuswiththe the preparative isolationoftanninsand Officer weare consolidatingourefforts in of HolgerGockowiakaspart-timeTechnical ade verygood a()c a()c a()(c) (a)(b) (a)(b)(c) (a)(b)(c) at pH3.6,(c)sameas(b)butinpresence ofSO (a) underacidicconditionsatpH1.5,(b)inmodelwine malvidin-3-glucoside andanthocyanin-derivedpigments. Figure 1.Colourproperties ofthegrapeanthocyanin could beenrichedintheaqueous the focusofstructure elucidationefforts, polar polymericpigments,whichare now important wastheobservationthatvery available grapemarc extracts.Equally anthocyanins from severalcommercially anthocyanins andcoumaroylated separations andoptimizedtheisolationof solvent step-gradientswithMLCCC (MLCCC). Forthefirsttime,weapplied layer coilcountercurrent chromatography continued toexplore thepotentialofmulti- anthocyanins and'pigmentedpolymers’,we For theisolationandpurificationof chemical specificityofthecalibrations. experiments are required todeterminethe sample preparation. However, more performing exhaustivepre-treatments or compounds inred fermentationswithout spectroscopy forrapidprediction ofphenolic these modulated mouth-feel properties. structural properties couldcontributeto concentration effects, molecularmassand polymers' andtoestablishwhether as reduced supposedly this area is required toconfirmthe in research futurecompounds, polymeric with them into lack of mouth-feel properties their carry might result couldsuggestthatanthocyanins tasted byourpanels.Whilethisveryinteresting grape seedandskinthathavebeen than thesmallestnativetanninsfrom apple, These pigmentedtanninswere lessastringent isolated byMLCCCforsensoryassessment. In addition,'pigmentedpolymers'havebeen stationary phaseuponMLCCCseparation. glucoside Maldivin-3- tringency of'pigmented anthocyanin-derived pigments 2 . pigments were almostinerttoSO demonstrated (Figure 1)thatthesewine extensive NMRexperiments.We also confirmed thestructuralassignmentby the synthesisedreference compoundswe thehelpof and oxidationreactions. With complex combinationofaddition,ringclosure the pigmentsfrom vinylphenolsinvolveda we coulddemonstratethattheformationof pigments havenowbeensynthesisedand For thefirsttime,thesemalvidin-derived in winesamples(Institutepublication#678). to detectnumerous potentialred pigments spectrometric experimentshadallowedus As reported previously, tandemmass 'pigmented polymers'andtannins Characterisation ofredwinepigments, ihD ae end (Or with DrJamesKennedy publications #660,686, spectrometry havebeenpublished(Institute of grapeseedtanninsbyelectrospray mass of ourstudyonthestructuralcharacterisation by massspectrometric analysis.Theoutcomes identity oftanninshasbeendemonstrated and inwinehavebeendeveloped, andthe and 'pigmentedpolymers'inseedextracts Methods toquantifythelevelsoftannins and productive research fortheTannin team. polymers' remained anarea ofhighpriority to characterisetanninsand'pigmented The developmentofmethodstomeasure and Analyt. Biochem.318,291-299). pKa ofmalvidin-3-glucosidebyelectrophoresis. M.E.; Jones,G.P. [2003]Chargeequilibria and wine pH(Asenstorfer, R.E.;Iland, P.G.; Tate, coloured isoformofmalvidin-3-glucosideat quinoidal isomercouldrepresent themajor and provided evidencethatanuncharged malvidin-3-glucoside atvariouspHvalues structures andcolourproperties of manuscript summarizingtheirresearch on – 46).Thesameauthorspublisheda Res.9,40 maturation. Aust.J.GrapeWine A duringred winefermentationand A.J.; Jones,G.P. [2003]Formationofvitisin (Asenstorfer, R.E.;Iland,R.G.;Markides, vinylphenols studiedbytheInstitute’s team malvidin-3-glucoside inpresence of required, similartothereaction of demonstrated thatanoxidationstepwas with fermentationtrials.Thisresearch Graham Jones(TheUniversityofAdelaide) project' membersRobertAsenstorfer and pigment vitisin-Awasstudiedby'Tannin formation oftheanthocyanin-derivedwine malvidin-3-glucoside withvinylphenols,the Similar totheabove-mentionedreaction of publication #725). during winemakingandageing(Institute most ofwhichare likelytobeformed stable andunbleachablered pigments, anthocyanins canbetransformedinto this research demonstratedhowgrape compared toanthocyanins.Insummary, coefficients inmodelwineatpH3.6when colour properties andenhancedextinction these winepigmentsshowedsuperior could beinstantlydecolorized,andthat anthocyanins suchasmalvidin-3-glucoside mediated bleachingwhilegrape 713). Incollaboration egon State 2 - tasted. Although the experimentswere mouth-feel characteristicswhena wine is components cancontributeparticular scarce knowledgeregarding what are veryvaluableadditionstoourpreviously la Recherche Agronomique inMontpellier Cheynier's group attheInstitutNationalde been carriedoutincollaborationwith Dr In summary, theinvestigations thathave rather thantheanthocyaninsthemselves. was duetominorimpuritiesofotherphenols perceived initiallywithlesspure fractions that thelowlevelofastringencyandfullness the solutionstheywere tastedin.Itislikely they influencethefullnessorviscosityof perceived asbitterorastringent,nordid purified anthocyaninfractionswere not study inMontpellier. Inbothstudiesthese ethanol aspartofapaired comparison undertaken attheInstituteandin5% part ofadescriptivesensoryanalysis hydrogen tartratesolutionatpH3.6)as (13% ethanolinsaturatedaqueouspotassium fractions havebeentastedinmodelwine were revisited andpurifiedanthocyanin The sensoryproperties ofanthocyanins than thetypeoflinkagebetweenunits. influence ontheirastringencyproperties subunits intanninscouldhaveabigger This result suggestedthatthenumber of linked through direct inter-flavan bonds. catechin oligomersofmDP3and9 astringency mid-waybetweenthatofnative degree ofpolymerisation5(mDP 5)had the syntheticcatechinoligomersofamean linkage through ethylbridges.When tasted, monomers inpresence ofacetaldehyde by oligomers were formedfrom catechin demonstrated thattannin-likecatechin other mouth-feelsensations.Itcouldbe quantifying astringency, and bitterness Ann Noble,time-intensitystudiesfor conducted, incollaborationwithProfessor in thepresence ofacetaldehydeand characterised catechinoligomersobtained publications #688,697and717,we anthocyanins assummarizedinInstitute sensory properties oftanninsand In additiontotheresults from studieson related polymers feel propertiesofcondensedtanninsand Understanding theastringencyandmouth- our knowledge. postulated, butneverbeenpublishedto anthocyanin andlargertanninshasbeen condensation products obtainedfrom Until now, theexistenceofsuchdirect tannins) amongstthe'pigmentedpolymers'. to eightcatechinsubunits(octameric rather largecondensationproducts withup we coulddemonstratethepresence of anthocyanins withtannins.Interestingly, of the products ofdirect condensation 'pigmented polymers'from PinotNoir were Analysis oftheMSdatarevealed thatthe and HPLC-MSexperimentsattheInstitute. were characterisedbyESI-MS,ESI-MS/MS samples prepared from PinotNoirwine University, USA)'pigmentedpolymer' choice ontannin and colourproperties was Jeff Eglinton),theinfluenceofyeaststrain (Dr PaulHenschke,DrEvelineBartowsky, In collaborationwithInstitutemicrobiologists astringency levelsingreater detail. tannin concentration,structure and will nowexplore thecorrelation between mouth-feel differences inthesewineswe To identifythebasisforperceived as methodstocharacterisetanninstructures. measurements oftanninsare asessential demonstrate thatprecise quantitative need tobeconfirmedbutnevertheless differences inastringency. Theseobservations fermenters at25ºC,whilelackingthe when compared towinesmadeinrotary featured enhancedextractionofphenolics made inPotterfermentersat25ºCalso at thisstageforthe‘Pottereffect’, aswines however, noconclusiveexplanationavailable by mechanicalstress onthesolids.There is, 2001 thatwere likelytoenhanceextraction centrifugal pumpsusedforpump-overin ‘Potter effect’ couldbelinkedtothe oversized properties. Onelikelyexplanationfor this types gavewineswithsimilarmouth-feel (25ºC) ferments,where thetwofermenter was notevidentforthehighertemperature fermented intherotary tanks.Thiseffect (drying, coarseandadhesive)thanthose the Potterfermenterwere more astringent Sauvignon,thewinesmadein Cabernet for allofthe18ºCfermentswithShirazor experiments hasbeencompleted.Notably, 2001 vintagecommercial-scale winemaking and descriptiveanalysisofwinesfrom the A sensorystudyinvolvingdifference testing properties ofwine andcoloursensory concentrations, Winemaking practicesthataffecttannin model systemwithtanninspresent. in theabsenceoftannins,butnot studied conferred thesensationoffullness the effect ofpolysaccharides,thetwotypes tannin-related astringency. regard With to tannins withanthocyaninscouldreduce structures, andwhethercondensationof incorporated intooligomericandpolymeric properties withthemwhentheyare anthocyanins carrytheirlackofmouth-feel in. Itremains tobeconfirmedwhether viscosity ofthesolutionstheywere tasted they didnotinfluencethefullnessor perceived asbitterorastringent.Inaddition, corresponding coumarates,were not anthocyanin glucosidesandtheir comparable size.Thegrapepigments, and astringentthanskintanninsof more as seed tanninswere perceived tannins and that smaller than concentration at agivenmass(ratherthanmolar) that largertanninswere more astringent most strongly, withthefindingsdemonstrating and concentrationinfluencedastringency As mightbeexpected,tannincomposition important advances. actual red wines,theresults pointtosome work isrequired toconfirmtheseeffects in carried outinmodelsystems,andfurther coarse 13 Research Teams’ report pursued. The wines from the joint 2002 members collaborated closely with the concentrate efforts on development of an commercial-scale winemaking trial comparing Institute’s Wine Microbiology and NIR NIR method for determining colour, total S. cerevisiae with S. bayanus have been teams to foster research synergies and to soluble solids (TSS) and pH in red grapes, analysed by HPLC and NIRS twice daily minimize trial-related costs. This approach that would be suitable for practical industry during fermentation until pressing, during has already been proven successful during use. Several reports and Roadshow MLF and after bottling. Statistical analysis the 2002 vintage, and it enabled us to presentations by the project team have of the HPLC data for malvidin-3-glucoside analyse all twelve fermenting tanks during introduced the potential use of NIR for red as the dominant grape-derived anthocyanin alcoholic fermentation on a daily basis. grape quality assessment and a detailed and 'pigmented polymers' formed during Comprehensive data were collected for article has been published in the 2003 fermentation has demonstrated that by phenolic composition by HPLC and NIR Australian & New Zealand Grapegrower & using rotary fermenters at 20ºC all wines spectroscopy. At present, all wines are Winemaker Technical Issue (Institute made with S. bayanus had less malvidin-3- undergoing malolactic fermentation and publication #730). Several large wine glucoside but more 'pigmented polymers', we will continue to analyze the samples at companies have now put this technology and also more tannins compared to wines bottling and after approximately twelve into practice. made with S. cerevisiae. Whether fermenting months with the sensory studies. with S. bayanus enhanced tannin extraction It has been proposed that transfer of the and/or resulted in enhanced formation of NIRS project: rapid instrumental techniques technology to the industry might best be tannin-like polymers remains to be clarified. achieved through direct commercialisation As these red wines have been made from Staff: Mark Gishen, Dr Bob Dambergs, Dr activities, and this is being carried out carefully randomized grapes under controlled Leigh Francis, Dr Wies Cynkar, Professor Peter under the responsibility of the Cooperative and replicated conditions, we are now Høj, Dr Elizabeth Waters, David Boehm, Les Research Centre for Viticulture's dedicated planning sensory studies with focus on the Janik, Dr Daniel Cozzolino commercialisation company, CRCV 'yeast-effect' on wine composition, wine tannin Technologies Ltd. CRCV Technologies Ltd., concentration and astringency properties. Collaborators: Chris Bevin, Audrey Lim with the assistance of a commercialisation (Hardy Wine Company), Russell Johnstone, consultant, have engaged in negotiations In addition to these commercial-scale Inca Lee (Orlano Wyndham), Dr Eric Wilkes with several instrument manufacturers in fermentations with S. bayanus, we studied (Beringer Blass), Dr Bruce Kambouris their management of trial commercial the chemical reactivity of anthocyanins (McGuigan Simeon) release of instruments provided with with grape-derived oenotannins in model calibrations developed by the Institute.

reactions in the presence of different SO2 Research into rapid instrumental methods and acetaldehyde concentrations. Addition has continued to concentrate on pursuing The consolidation of calibrations developed

of both SO2 and acetaldehyde to the promising technique of near infrared for the analysis of grape berry colour, total anthocyanin/tannin mixtures resulted in (NIR) spectroscopy. This analytical approach soluble solids and pH by NIR scanning has reduced formation of 'pigmented polymers' has been shown from earlier work to be continued, involving study of as wide a when compared to model reactions with capable of providing very fast, low cost range of samples as possible. The continued acetaldehyde alone, but still enhanced analyses of a range of parameters important cooperation of industry partners has meant 'pigmented polymer' formation when to commercial wine production. Spectroscopic that approximately 3000 berry samples from compared to the model reactions between techniques offer the potential to simplify the 1999, 2000, 2001, 2002 and 2003 anthocyanins and tannins in presence of and reduce analytical times for a range of seasons from a wide range of growing

SO2 alone. In summary, acetaldehyde was grape and wine analytes. It is this aspect, regions and representing several red identified as one likely yeast metabolite together with the ability to simultaneously varieties (but predominantly Shiraz and that could contribute to enhanced measure several analytes, which was the Cabernet Sauvignon) have been analysed formation of 'pigmented polymers' from impetus for developing NIR methods. by the conventional laboratory methods anthocyanins and tannins, and we are now and scanned with a research grade NIR aiming to measure acetaldehyde on a In response to the wine industry's need for instrument located at the Institute. The regular basis during red wine fermentations rapid analytical methods for the berry samples were scanned with no with the help of a GC-MS method that has determination of objective indicators of preparation other than homogenisation recently been validated. grape quality, a decision was made to using a high-speed laboratory homogeniser.

Our main objective with the on-going 2003 Tannin trial is to study the impact of tannin additions (200 ppm commercially available 'colourless', proanthocyanidin-based grape seed tannins) on red wine phenolic composition, tannin concentration, colour and mouth-feel properties. With the current trial we are aiming to demonstrate the effects of adding the oenotannins pre- or post-fermentation to Potter fermenters. We also expect to ascertain whether our existing analytical methods are specific enough to identify the molecular basis for the perceived colour and mouth-feel changes. The grapes for the 2003 large scale winemaking trials of the 'Tannin project' and the Institute’s microbiology team have been provided by Banrock Station and the excellent support from Ben Vagnarelli and his team is gratefully acknowledged. After crushing of the grapes in the Hickinbotham Roseworthy Wine Science Laboratory, the Tannin team Daniel Cozzolino Wies Cynkar Les Janik Bob Dambergs Research Teams’ report Teams’ Research 14 It has been found previously that, while the sub-sampling, weighing, addition of an several compact, fast, diode-array calibrations appeared to hold across extraction solvent, mixing during extraction, spectrometers including the Zeiss Corona. seasons and a calibration could be developed and finally centrifugation, and is multitasking. Since the 2001 vintage, the NIR team has that will give acceptable results for samples Following extensive evaluation at the Institute been collaborating with the Tannin Project from multiple regions or from several of the commercial system, the unit has been team in their investigations of process scale varieties, best accuracy seemed to be found to perform well in all tests, including fermentations. The main objective was to obtained from restricted sample sets (e.g. those with sets of grape berry samples with examine the potential of NIR spectroscopy same vintage, region or variety). It has challenging physical properties. In recent to predict the concentration of phenolic been shown that a regression method testing, 156 grape samples were processed compounds and monitor the extraction known as LOCAL could dramatically reduce and analysed. The data from the automated and evolution of these compounds during the prediction error for grape colour from system compared well to the results obtained red wine fermentation. If successful, NIR large, diverse data sets, but a simple from the manual method in terms of spectroscopy techniques may offer potential method may be to tailor standard partial accuracy and precision. No serious practical as a rapid, low cost and non-invasive tool least squares (PLS) calibrations for samples deficiencies were observed in the testing, for monitoring the fermentation process. of similar colour values. Table 2 compares with aspects such as ease of access to the The preliminary results from the 2001, 2002 calibration statistics for the full 1999 to parts of the unit, ease of cleaning and and 2003 winemaking trials conducted at 2001 dataset with subsets selected by control of the processing being satisfactory. the Hickinbotham Roseworthy Wine colour range, irrespective of vintage, grape The unit could be configured to act as an Science Laboratory showed that NIR variety or growing region. A mid-range automated homogeniser, with no further spectroscopy could predict the colour set (colour between 0.5 and 1.5 processing, to allow NIR scanning of the concentration of malvidin-3-glucoside (the mg/g) had a lower standard error of cross samples only (i.e. with no reference analyses), major anthocyanin in grapes), ‘pigmented validation (SECV) than the full data set and in which case the system could process polymers’ and tannins in Cabernet Sauvignon the equation developed with this mid- approximately 112 samples/8 hour day. For and Shiraz wines during fermentation. range set had an increased standard error full processing (including extraction and However, the specificity of the calibrations of prediction (SEP) when predicting the clarification) with one operator carrying out developed must be confirmed, as there are extreme range samples. Note also that an other analytical tasks on the samples, 48 many simultaneous changes occurring during equation developed with 20% of the mid- samples would be the likely maximum that fermentation. Results from a calibration for range set could predict the remaining 80% could be processed in an 8-hour period. malvidin-3-glucoside in red wines from 2001, with better accuracy than the full data set, 2002 and 2003 fermentation trials are thus with a large dataset, analytical workload The project has continued to investigate shown in Figure 2. can be reduced by using an NIR method. the effect of sample presentation in NIR Sample storage and processing are analysis, comparing homogenised grapes The authentication or identification of food important issues that require consideration with whole red grapes. Initial work by the is emerging as one of the most important when performing analysis on grape project team on the prediction of quality areas in order to comply with food industry samples. The effect of freezing and parameters in red grapes using NIR began regulations and standards and to address homogenisation of red grapes on the with the scanning of homogenized grape consumers' concerns. Since the early 1980s, determination of total anthocyanins, TSS and pH was determined recently. Three different commercial homogenisers were Table 2. Calibration statistics for NIRS measurement of colour (mg/g) in samples from multiple vintages, regions and varieties separated by colour range; midrange: 0.5–1.5 mg/g; extreme-range: >1.5, <0.5 mg/g used for sample preparation. The effects of freezing, homogeniser type and storage 2 Calibration set N Validation set SECV SEP R time on the accuracy and recovery of the laboratory determination and NIR All samples, 1999 - 2001 2287 - 0.13 - 0.92 prediction of colour, TSS and pH are being evaluated. Preliminary results have mid-range colour 1618 - 0.09 - 0.89 indicated that neither freezing of samples mid-range colour 1618 extreme-range colour - 0.16 0.92 prior to homogenisation, nor the type of homogeniser had any significant effect on 20% subset of mid range 324 0.09 - 0.91 the accuracy of laboratory determinations 20% subset of mid-range 324 80% subset of mid-range - 0.10 0.87 of colour and TSS. However, as one might expect, there was an effect of freezing on Notes: N is the number of samples, SECV is the standard error of cross validation, SEP is the standard error of prediction, pH. This study is continuing with further and R2 is the coefficient of determination. samples remaining in frozen storage for later analysis to investigate any long-term stability effects as empirical evidence samples using a research grade laboratory the development of multivariate statistical indicates long-term freezing at -20°C can NIR spectrometer, the FOSS NIRSystems techniques, personal computers, and new lead to colour loss. This study will provide 6500. However, the recent availability of instrumentation have resulted in the the basis for updating the protocols for the faster scanning Vis/NIR spectrometers with development of IR methods that offer the analysis of red grapes by both reference alternative optical configurations has possibility to analyse more samples than ever and NIR methods. provided the possibility of presenting the before and acquire large amounts of data. grape samples to the instrument without Spectroscopic techniques such as NIR or The Institute has been involved in the testing the need for homogenisation. Simplifying Fourier transform infrared (FT-IR) have proved and assisting in the development of an the sample presentation for NIR prediction to be a suitable tool for authentication automated grape sample preparation system of colour, TSS and pH could dramatically with demonstrated applications including ‘Bioprep5 Robotics’, which has been released increase sample throughput. Furthermore, discrimination between coffee varieties, commercially by ARRM, a local Adelaide this mode of presentation might even offer honey adulteration, and beef contamination company. The unit is designed primarily to the potential of scanning whole, single and adulteration. The project team has process grape samples for colour analysis, berries. Preliminary investigations for whole commenced work using NIR spectroscopy whether by NIR or by the reference laboratory grape berry presentation were promising, to discriminate between wine varieties and method, but it can be put to other uses. indicating that NIR may have potential for blends. Preliminary investigations have The benchtop-sized unit is fully automated, use at the weighbridge or for in-field analysis. been undertaken into the discrimination of carrying out grape sample homogenisation, These studies have been performed using commercial samples of two white wine Research Teams’ report Teams’ Research 15 16 Research Teams’ report The project teamhascontinuedwith particularly red grapes. with mechanicallyharvestedfruit, visual inspection,butthiscanbedifficult weighbridge wouldnormallybedone by assessment forfungalinfectionat the to determinesuitabilityforwinemaking. highlight suspectfruitforfurtherdetailedanalysis the weighbridgetoprovidea'go/no-go'test possible todiscriminatefungalinfectedfruitat The implicationofthisworkisthatitmaybe The UniversityofAdelaide. with theFungalPathologyLaboratoryof This workisbeingdoneincollaboration secondary infectionbyotherorganisms. removing theproblem ofconfusionwith control ofinfectionatvariouslevelsand fruit infectedinthelaboratory, allowing has theadvantagethatitcanbedoneon Botrytis- Similar workisalsoinprogress with fungal constituents. induced grapecompositionalchangesfrom now focusondifferentiating fungus- at leasta10%infectionlevel.Work will classify powderymildewinfectiondownto changes were usedtodetectandcorrectly of infection(Figure 4).Thesespectral classified intoanumberofdifferent levels changes couldalsobeobservedinfruit homogenised leaves.Distinctspectral demonstrated usingasmallsamplesetof principal componentanalysis(PCA)was control anderinoseinfectedleavesusing sporulating lesions,assessedvisually)from mildew leafinfection(earlylesionsand Discrimination oftwostagespowdery obtained from NIRscansofinfected samples. assay –thesecanbeusedtoverifyresults has mildew andCSIRO a has developed The F in Canberra. University ofAdelaideandCSIROPlantIndustry with theFungalPathologyLaboratoryofThe powdery mildewiscontinuingincollaboration spectroscopy fordetectionof An evaluationofthefeasibilityNIR during laboratoryfermentations. monitor ethanol,sugarsandothercompounds suitability ofIRspectroscopic techniquesto Microbiology teaminorderWine toassess the also beenplannedincollaborationwiththe strains hasbeenexamined.Furtherwork spectroscopy todiscriminateandidentifyyeast team, thepotentialofNIRandMIR In collaborationwiththeMolecularBiology this technique. red wineswillalsobeinvestigatedusing classification specificity. Itisproposed that the influenceofwinematrixon work iscontinuinginorder tounderstand two varieties,asshowninFigure 3.Further NIR spectrashowsdiscriminationbetweenthe Smyth. Principalcomponentanalysis(PCA)of flavour compounds,conductedbyHeather from aPhDproject onidentifyingwhitewine varieties (Chardonnay andRiesling),sourced infected fruit.The DNA assayforpowdery ungal PathologyLaboratory developed anELISA Botrytis Botrytis work Grape and

Intensity -0.1445 -0.1176 -0.0908 -0.0640 -0.0371 -0.0103 0.0185 0.0434 0.0702 0.0970 Malvidin-3-glucoside predicted by NIR (mgL-1) 2 5 8 1 4 8 1 4 7 0 4 7 0 3 6 0 3 6 9 10281060 996 964 932 900 868 836 804 772 740 708 676 644 612 580 548 516 484 452 420 100 150 200 250 300 350 400 50 PCA 4 0 (yellow), and61-100%(lightblue)powderymildew infectedChardonnay grapes Figure 4.Firstderivativespectraof uninfected (darkblue),1-10%(red), 11-60% 0101020203030400 350 300 250 200 150 100 50 0 Chardonnay (purple)wines spectra, showingdiscriminationbetweenRiesling(blue)and Figure 3.Principalcomponentscores forPC1andPC4ofNIR PLS: partialleastsquares) ferments (n:numberofsamples;SEC:standard error ofcalibration; analyisis (HPLC)valuesformalvidin-3-glucosideinred wine Figure 2.Therelationship betweenNIR-predicted andreference 3 PLSfactors R n=545 SEC=26.60 mgL 2 =0.92 Malvidin-3-glucoside byHPLC(mgL -1 Wavelength -1 ) PCA 1 investigations on the application of the MIR Grape composition and wine flavour been unable to identify the fifth. The sixth (mid-infrared) technique, used a rapid and component, which was also the most simple sample presentation method called Staff: Dr Mark Sefton, Professor Peter Høj, potent of the six in the GC/olfactometry attenuated total reflectance (ATR). With this Dr Leigh Francis, Dr Markus Herderich, assessments, was tentatively identified as technique, the grape homogenate sample Dr Elizabeth Waters, Dr George 2,3,6-trimethylphenylbutadiene (TPB) from is simply smeared onto the ATR crystal and Skouroumounis, Dr Alan Pollnitz, Dimitra its mass spectrum. An authentic sample of scanned, thereby eliminating many of the Capone, Mango Parker (nee Ranzijn), Kevin TPB was synthesised and this has problems associated with the use of sample Pardon, Kate Lattey, Heather Smyth, Dr confirmed the assignment. TPB has not cuvettes, which require a very small pathlength Gordon Elsey, Carolyn Puglisi, Aggie Janusz, been previously reported as a grape or in the MIR spectral range. Scanning of the Merran Daniel wine component. The threshold of TPB in homogenates of grape samples from the a white wine was determined to be 40 2002 vintage was conducted on both the This project has the long-term objective of ng/L and common descriptors used by our NIRSystems 6500 (NIR) and an FT-MIR understanding how the combination of panel for this compound was 'floral', spectrometer. Comparative calibration viticultural and winemaking practices 'geranium' and 'tobacco'. This makes it statistics for the two techniques suggest that determine the aroma and flavour one of the most potent wine flavour MIR may have superior predictive capability characteristics of wine and is divided into compounds known. An analytical method for pH and TSS, but is less accurate than three broad areas: the identification of for TPB using d8-naphthalene as internal NIR for colour. However, the best prediction grape-derived wine components that affect standard was developed and several wine for colour was achieved using a combination wine aroma and flavour; studies of the samples were analysed for TPB, which was of MIR and NIR. Work is continuing with formation and degradation of these wine observed in four out of the five white both of these techniques using samples flavour constituents including the wines examined at a concentration of 50- from the 2003 vintage. identification of their grape-derived 210 ng/L. It was not observed in the four precursors; and finally, studies on how red wines examined. A more sensitive and In late 2002, the project, together with the viticultural variables affect the production accurate analytical method for TPB using Institute’s Analytical Service, commenced of wine flavour compounds and their stable isotope dilution analysis is well evaluation of an FTIR-based spectrometer precursors in the grape. advanced – a deuterium labelled analogue (FOSS WineScan) for assessment of its has already been successfully synthesised in capability in the routine analysis of wines. Analysis of wine components and their precursors a multi-step pathway. Wine samples, and corresponding analytical data, were used for re-calibration of the Analytical methods for 33 important Formation of important wine flavour compounds instrument, and the resulting calibrations fermentation esters, acids and alcohols using from grape-derived precursors used to enhance the ‘global’ calibrations their deuterium labelled analogues as provided with the instrument. Although the internal standards have been fully validated. Riesling Acetal is a relatively weak-smelling calibrations provided by FOSS allow analysis Most of these compounds can be analysed grape derived norisoprenoid that is formed of up to 18 wine properties, the current in a single GC run. The methods are now along with the more potent 1,1,6- study is examining only nine of these viz. being applied to other projects under this trimethyl-1,2-dihydronaphthalene (TDN) alcohol, pH, density, glucose/fructose, total project Grape composition and wine flavour from grape derived precursors. In the short acidity, volatile acids, acetic acid, citric acid and Yeast flavour and fermentation activity. term, these two products are formed in and malic acid. Preliminary results for the competition, and we considered that first six months of operation have been very A sample of deuterium-labelled citronellol factors such as the position of glycosylation encouraging and were presented in New has been prepared, and an analytical on the polyfunctional precursors could Zealand in May 2002 at the FOSS Directions method for this monoterpene has been steer the conversion towards or away from 2003 conference. developed. The method is now being TDN formation. Recent hydrolytic studies applied to sensory study of Riesling and by us have shown that Riesling Acetal can The project team has also been working with Chardonnay wines (see below). in fact be completely converted to TDN at other groups in the application of NIR to wine pH, probably over a period of several predict red grape quality parameters (colour, Large scale syntheses of the potent wine years, and that this will contribute to a TSS and pH) in relation to their research flavour compound ‘wine lactone’, its gradual increase in TDN in bottled wines projects. The objective was to reduce the deuterated analogue, and the naturally over this period. As expected for acid cost of analysis and the necessity for wet occurring (optically active) form have catalysed transformations, the formation of chemistry. Such work is underway with recently been completed. These standards TDN from Riesling Acetal was approximately groups at CSIRO (Land & Water's Precision will be used to develop analytical methods twice as fast at pH 3.0 as at pH 3.2. Agriculture group) and The University of for wine lactone in wines and in model Adelaide (Roseworthy Campus). Strengthening media – the latter is needed for studies on Four isomeric allenic diols, all potential of relationships with wineries has resulted the formation of wine lactone from grape- damascenone precursors, have been from both the development of some training derived precursors. The optically active synthesised and purified. The availability of material in chemometrics and multivariate form will be used in the sensory studies of these authentic samples allowed us to analysis for wine industry personnel, as well Riesling and Chardonnay (below). prove that they were intermediates in the as exchanging experiences through interaction formation of the important wine aroma with other industry staff working in the area Identification of new wine flavour components compound damascenone, as had been of IR spectroscopy. postulated previously by us. Hydrolytic It is proposed that an 'IR spectroscopy users Hydrolysates of a grape-derived studies into the formation of damascenone group' be established bringing together the glucoconjugate purified by MLCCC have have been conducted for all four isomers. Institute’s NIR team, and other researchers been analysed by GC/olfactometry/mass At room temperature and pH 3.0, each and practitioners involved in the development spectrometry and these chromatograms isomer gave damascenone in high yield and application of IR spectroscopy in the compared with those of hydrolysates of the within a few days. Surprisingly, no trace of research and industrial communities. This will total glycoside pools isolated from grapes. 3-hydroxydamascone was formed from be an important step in further strengthening The former are much simpler than the these compounds. 3-Hydroxydamascone is the links between the Institute and industry latter, and it has been possible to assign normally found in grapes and wines at in the application of the IR technology in structures to five out of the six odorants much higher concentration than is the Australian wine industry. detected in the chromatogram. Four of damascenone. These findings have led us the odorants have previously been reported to reevaluate the mechanism by which

as wine components, and we have so far damascenone is formed in young wine. report Teams’ Research 17 18 Research Teams’ report dried rose character, andwines withstronger perfumed aroma compared tothosewitha samples witharelatively strong floral intensities ofspecificattributes,such as group there were wineswithvaried each broad and toastyattributes.Within like, andolderwineswithlime,kerosene those thatwere tropical fruit/passionfruit- dominated byfloralandlemonaromas, samples: thoseyoungwineswhichwere wines, there were three main groups of sensory dataindicatethat,fortheRiesling Chardonnay wineshasbeencompleted.The to two-yearoldcommercial unwooded of 20commercial Rieslingwinesand20one A sensorydescriptiveanalysisstudyonaset properties ofwine Relationship betweencompositionandsensory investigated inthenearfuture. determine itsconcentrationinwinewillbe a verylowaroma threshold. Amethodto investigation. Oneofthesecompoundshas synthesised, withafourthcurrently under present, three suchcompoundshavebeen contributors towinearoma andflavour. At see ifthesecompoundsare important is underway. Thisworkisbeingdoneto analogues ofcommongrapecomponents A studyintotheformationofsulfur these compoundsingrapesandwines. develop stableisotopedilutionassaysfor progress. Theseanalogueswillenableusto deuterium labelledanaloguesisalsoin these importantthiols).Asynthesisof conjugates are grape-derivedprecursors of flavour release duringfermentation(cysteine fermentation activity being usedinthe been synthesised.Thisconjugateisnow cysteine conjugateofonethesehasalso have beenprepared, andasampleof progress. Two ofthethree targetcompounds in manyred andwhitewinestylesisin thiols thatare keyflavourimpactcompounds The synthesisofagroup ofgrape-derived precursors discussedabove. damascenone formationfrom theallenic has beenidentifiedasaminorbyproduct of reactions were synthesised,andoneofthese wine. Theexpectedproducts ofthese concentration normallyseeninageing the more rapiddecrease indamascenone expected, andalonecannotaccountfor concentration inthesereactions waslessthan examined. Thedecrease indamascenone potentially reactive compoundswere as winesage)hasbeencompleted.Ten components (i.e.thelossofdamascenone of damascenonewithcommonwine A preliminary thereaction studyconcerning formation ofdamascenoneinnature. helped clarifyourunderstandingofthe within theprecursor molecule,andthishas the relative reactivity ofthefunctionalgroups studies hasgivenusavaluableinsightinto the intermediatesformedinhydrolytic Analysis bychiralgaschromatography of Yeast flavourand research program on of All fourpossibleisomersoftheglucosides Kevin Pardon Dimitra Capone,KerryWilkinson, Staff: wine composition The influenceofoakcooperageon data analysishasalready beenundertaken. in thenearfuture, andsomepreliminary all ofthesewineswillbelargelycompleted isoprenoids. Thecompositionalanalyses of the majormonoterpenesandnor- have already beenquantified,aswell as concentration ofesters,alcoholsandacids completed. FortheRieslingwines, once allchemicalanalyseshavebeen and Chardonnay wineflavourattributes compounds andtheintensityofRiesling concentration ofparticularvolatile made intotherelationships betweenthe The datasetshouldenableinsightstobe herbaceous characters. showed prominent passionfruitand A subsetoftheseyoungerwinesalso dominated byestery, citrusandfruitynotes. old wines,whiletheone-yearwineswere two-year the in descriptors were prominent honey, butterscotch,w honey-like aroma. FortheChardonnay wines, hydrolyse tooaklactoneduringwine would not,therefore, beexpectedto glucosides andthegalloylatedderivative slowly togiveoaklactone.Boththese wine pH,andevenat100˚C,reacted only The glucosideswere relatively stableat galloylated analogue. useful modelstostudythereactivity ofthe The simpleglucosides,therefore, serveas glucoside already identifiedinoakwood. structurally similartoagalloylated potential precursors ofoaklactoneandare These compoundsare considered tobe synthesised andobtainedinpure form. cis -and Dr MarkSefton,AlanPollnitz, trans -oak lactonehavebeen oody andspicy Gordon Elsey&HeatherSmyth Gordon and theshavingswere thenheatedto235˚C lactone were absorbedontooakshavings mixtures of two forms(i.e.theracemate),orevenon has beencarriedoutonmixtures ofthese sensory workon non-natural opticalisomer. Previous identical isomerof have beenabletoresolve thenature- of theglucosidesoaklactones,we As aresult ofourobtainingpure isomers have beenreported bymanyauthors. oak lactoneinasaresult ofheating lactone duringbarrel toasting.Increases in glucoside canactasprecursors tooak glucosides andprobably alsothegalloylated observed. Thisdemonstratesthatthesimple of oaklactonesintheshavingswere in anoven,substantialincreases intheamount nature-identical isomerof The sensorydetectionthreshold ofthe When theglucosidesof been separatedfrom theextract. hydrolytic means,oncetheshavingshave model wineextractsofoakshavingsby that cancontinuetobegeneratedin for theadditionalamountsofoaklactone wine. Suchcompoundscannotaccount glycosidase activity)were present inthe enzymes (ormicroorganisms with conservation, unlessactiveglycosidase different. We are nowinapositionto descriptors fortheseisomerswere also sensory panellistsindicatedthatthe aroma intensity, butinformalcommentsbythe these twoisomersdiffer intheirodour natural isomerof86µg/L.Notonly did threshold for thenon- detection a obtained below 10µg/L.Thesamesensorypanel to detectthenaturalisomerataconcentration mixture. Someindividualpanellistswere able of 90g/Lpreviously reported fortheracemic 23 µg/L.Thisismuchlowerthanthevalue trans (which istentimesmore potentthanthe form) inaneutraldrywhitewinewas cis- and cis cis- -oak lactoneinwine trans- oak lactonefrom its cis- oak lactone. cis- and oak lactone trans- oak carry out formal sensory descriptive analysis In the area of Brettanomyces research we yeast, that is, having a lower temperature on wines spiked with the nature-identical have fully validated SIDA methods for 4- range for growth and fermentation, our isomer of cis-oak lactone in order to determine ethylguaiacol, 4-vinylguaiacol, 4-vinylphenol research, and that of others, is showing its sensory properties more rigorously. and isovaleric acid. These methods supercede that S. bayanus has other properties of less reliable and more time-consuming oenological interest. These properties are A previous report by American researchers methods previously used. being investigated both in the laboratory hypothesised that cis- and trans-oak lactones and by collaboration with winemakers occur in equilibrium with their open chain Yeast flavour and fermentation activity working in the commercial environment. forms, and that conflicting data on oak The laboratory-based work has the aim to lactones in oakwood in the literature was a Staff: Dr Paul Henschke, Jeff Eglinton, Dr identify oenological differences with result of the failure to take such equilibria Eveline Bartowsky, Dr Peter Costello, Jane S. cerevisiae, the universal reference yeast, into account. We have, therefore, McCarthy, Kate Howell, Professor Peter and to gain an understanding of their determined the acidity constants (pKa) of Høj, Dr Mark Sefton, Dr Alan Pollnitz, physiological and biochemical basis so that the open chain forms, their rates of Tracey Siebert, Dimitra Capone, Gayle winemakers can benefit. By working closely lactonisation at different pH values and Baldock, Dr Diego Torrea with winemakers, we can develop a better ethanol concentrations, and the extent of understanding of the commercial potential lactonisation under these various conditions. This project has the long term aim to of various yeast properties, and to focus our The data show that, at wine pH (3.0 – 3.8), understand the role that yeast have in research effort more closely. Progress, which oak lactones exist almost entirely in the modulating wine sensory characteristics so has also benefitted greatly from collaboration lactone form at equilibrium, in contrast to that winemakers can more effectively with the Wine flavour, Polymers and the earlier hypothesis. At the highest pHs exploit the benefits of yeast for commercial Sensory teams in the Institute, has been studied, however, the open chain form of gain. During the past year, work has summarised in previous annual reports. the cis-isomer lactonised relatively slowly. focussed on model fermentation studies At pH 3.8, 50% lactonised after 16 days at related to understanding the role of yeast Our work is currently focussed on the room temperature, and at pH 5.0 in model in development of red wine colour, volatile and non-volatile metabolites of spirit, only a few per cent lactonised after commercial trials with experimental S. bayanus that have potential winemaking several days. The trans isomer lactonised Saccharomyces bayanus strains, and benefits. Higher amounts of glycerol and some 14 times faster than the cis isomer applying the stable isotope dilution assay lower amounts of acetic acid (a small under all conditions. As it is possible some suite to experimental Chardonnay wines to reduction in ethanol is also observed) oak lactone may originally be formed in wood understand better how fermentation together with increased succinic acid levels in open chain form, these data indicate the conditions affect aroma profile. are typically formed at usual winemaking need for suitable soaking times and low pHs Dr Diego Torrea, a recent PhD graduate temperatures (Institute publications #632 of wine or aqueous extracts when preparing from Professor Carmen Ancin’s laboratory, and 647). S. bayanus is typically competitive extracts of oak shavings for analysis. Navarro University, Pamplona, Spain, has and dominates fermentation (Institute been awarded a two-year fully funded publication #707) and although it may Chemical analysis of industry technical problems post-doctoral scholarship from the Spanish exhibit a slower rate of fermentation Government to work on aspects of the role towards the end of fermentation, the Staff: Dr Mark Sefton, Yoji Hayasaka, Gayle of yeast in development of wine aroma. wines generally have low residual sugar. Baldock, Dr Alan Pollnitz, Dr Markus Herderich, Diego already has considerable experience The prolonged fermentation times may be Tracey Siebert, Dimitra Capone, Kevin Pardon on chemical analysis of wine aroma and linked to improved palate structure and collaboration with Professor Ancin’s team weight noted by a high proportion of Problem solving work and research into the will greatly benefit this project. Dr Peter industry trials. Work is currently in progress formation of 4-ethylphenol and 4- Costello has resigned from the Institute to define better this important wine ethylguaiacol in wines is carried out by the (July 2003) to take up a position as the attribute and to establish its chemical basis. wine chemistry team in collaboration with, National Technical Manager with Lallemand and under the direction of, the Industry Australia. Peter brought a wealth of research In white wines, the fruity aroma of Services section. Accordingly, the results of and industry experience to this project and S. bayanus is often described as being this work are given in the report by Industry contributed strongly to chemical analysis as more complex than that achieved with Services. A main role of the project Chemical well as fermentation microbiology. As in S. cerevisiae (Institute publications #632, analysis of industry technical problems is to previous years, this project has benefitted from 647, 679 and 693). For example, Chardonnay provide analytical support to these areas. enthusiastic collaboration by a small group wines made with S. bayanus are typically of winemakers in trialling experimental rated higher in the attributes cooked Analytical methods for taint compounds in Institute yeast and providing wines and orange peel, botrytis/apricot, yeasty, wine continue to be developed. We have technical information. This close association savoury, nutty and aldehyde and lower in fully validated methods for compounds is important in maintaining a commercial the estery, pineapple, peach and citrus derived from petroleum products, including focus on project work and maximising the attributes compared with those wines made toluene, styrene, C2 alkyl benzenes (e.g. benefit of limited resources. with S. cerevisiae. Commercial Chardonnay xylenes), C3 alkyl benzenes (e.g. wines have shown some rose aromas trimethylbenzenes), C4 alkyl benzenes (e.g. Saccharomyces bayanus – a research tool for which are characteristic of the compounds, tetramethylbenzenes), naphthalene, understanding the role of yeast in modulating 2-phenylethanol and 2-phenylethyl acetate, methylnaphthalenes, dimethylnaphthalenes wine sensory characteristics and a fermentation with much less obvious aldehyde-like or and trimethylnaphthalenes. A method for yeast with commercial potential oxidised aromas. S. bayanus appears to 2-aminoacetophenone (reported to be produce a different profile of fermentation responsible for the so-called ‘untypical Saccharomyces bayanus is a species of the esters and alcohols, and work is in progress ageing’ taint encountered in some German Saccharomyces sensu stricto group to identify the key impact compounds white wines) using stable isotope dilution together with the globally favoured wine associated with S. bayanus wine aroma and analysis (SIDA) has also been validated and yeast, S. cerevisiae. The consequential to establish how winemaking conditions is being used to survey Australian wines. A sharing of many qualitatively similar affect them. This work is benefitting from series of deuterium-labelled chlorophenols physiological and biochemical attributes, the suite of stable isotope dilution assays have been synthesised and are being used facilitates deployment of S. bayanus in recently developed by the Institute’s Wine to develop methods for their unlabelled wineries with familiar S. cerevisiae flavour team led by Dr Mark Sefton (see analogues. These methods are well advanced. technology. In addition to being a cryotolerant previous annual report). Research Teams’ report Teams’ Research 19 20 Research Teams’ report ‘pigmented polymers.’ were requiredformaximalformationof yeast cells,activelyfermentingcells ‘pigmented polymers’intheabsenceof Despite some‘chemical’formationof being convertedinto‘pigmentedpolymers.’ suggesting thatsomeoftheanthocyaninswere in theconcentrationofanthocyanins, polymers’ wasaccompaniedbyadecrease tannins andyeast.Theformationof‘pigmented solutions containinganthocyaninsand/or ‘Pigmented polymers’wereformedinmodel has beenprepared todisseminatethis of red winepigments. Adraftmanuscript mediated reactions involvingtheformation the modelsystemforstudyof yeast- This studydemonstratedtheusefulness of of tannins,anthocyanins,andyeastcells. supplemented withdifferent combinations consisted ofasyntheticmediumthatwas in thefirstinstance.Themodelsystem system inthelaboratory, and investigated usingamodelfermentation polymers’ duringfermentationhasbeen basis forthedevelopmentof‘pigmented wines (Institutepublication#729).The of ‘pigmentedpolymers’inthe or Sauvignon winesmadewith major difference betweenCabernet previous annualreports. Forexample,a red winehavebeensummarisedin modify thecolourdensityand/orhueof yeast speciesandstrainshavetheabilityto The results ofstudiesshowingthat some red winecolour-modelfermentationstudies Probing theroleofyeastindevelopment reported below. yeast indeterminingwinecolourare Aspects ofthisworkrelated totherole of Tannin Microbiology teams. andWine a largeresearch effort bytheInstitute’s its basisinwinephenolics,isthesubjectof important winemakingattribute,whichhas (Institutepublication#729).This journal and recently publishedinanindustry 2003, ACSproceedings, inpreparation) conferenceinternational (Eglintonetal. annual report, presented atarecent measurements asreported intheprevious been supportedbyanalytical resulting from theuseof The observeddifference inred winecolour the isotope dilutionassaysasisbeingdonefor is beinginvestigatedwiththestable chemical basisforthesearoma differences more complexaromas andflavours.The showed leaner, slightlylighterfruitwith wine madewith and fresh fruitaromas andflavours, AWRI 838exhibitedbrighterpurplecolour Sauvignon winemadewith undertaken sofarsuggestthatCabernet aroma, palateandcolour. Thelimitedtrials red winemakingpotentialwithrespect to Saccharomyces bayanus S. bayanus S. bayanus was thehigherconcentration white wines. S. bayanus appears tohave S. bayanus S. cerevisiae AWRI 1375 S. cerevisiae S. cerevisiae S. bayanus has Diego Torrea, PaulHenschke&EvelineBartowsky expression ofvarietalcharacter. More recent notes whileretaining fruitinessandgood tendency to general, winearoma isdistinctivewitha complex aroma noteswere observed. In with of thefruityaromas typicallyassociated aldehyde, yeasty,botrytis/apricot, savoury, nutty notes, suchas typically notedagreater diversityofaroma In theearlywhitewinetrials,winemakers Sauvignon. Noir andCabernet included Semillon,SauvignonBlanc,Pinot predominantly onChardonnay, buthave blends. Thetrialshavefocussed led towinebeingusedincommercial extensive triallingbytwocompanieshas October 2001.Overrecent vintages, more IndustryTechnicalWine Conference in workshop conductedatthe11thAustralian withnon-conventionalyeasts Winemaking Institute Roadshowseminarsandthe andvia winemakers inindustryjournals these trialshavebeencommunicatedto since 1999.Findingsarisingfrom several of trials rangingfrom barrel to10hLferments and NewZealandhaveconductedwinery 1375. SometencompaniesinAustralia Institute’s winemakers intheuseoftwo Considerable interest isbeingshownby fermentation strains Winery experiencewithSaccharomycesbayanus Eglinton andDrMarkusHerderich. Tannin teamsunderthesupervisionofJeff Microbiology and between theWine This workcontinuesasacollaboration manuscript fortheconference proceedings. Herderich andincorporatedinthe USA –SeeAppendix1)byDrMarkus conference (AmericanChemicalSociety, study havebeenpresented ataninternational information toindustry. Someaspectsofthe S. cerevisiae S. bayanus in additiontoalowered impact okdfruits/oldrose/savory/nutty cooked cooked orangepeel, strains. Occasionally, more strains, AWRI 1176and and being addressed asreported below. culture availability. Thisissueiscurrently constrained bythelackofcommercial starter trialswithred varietieshavebeen Winery of buildingwinestyle. an additionalblendingoptionforthepurpose with thesetrialssee noted. Manyofthewinemakersinvolved trials, betteroakintegrationiscommonly comparison with creamier texture, impartedby palate fullnessandrichness,oftena consistentlyreport weight. Winemakers trials are focussingonpalatestructure and Institute’s from alaboratorysource. Currently, the time/resources required toculture them of suitablestartercultures andthe companies isbeingrestricted byavailability winemaking trialsplannedbyseveral are needed.However, themore extensive for whichonlysmallvolumestartercultures strains wasconfinedmainlytobarrel trials Initial interest intheInstitute’s culture development starter Saccharomyces bayanus useful form. availability ofthesestrainsinacommercially conclusion tothisproject willbetheready 1375. Theoutcomeofasuccessful produce ADWYfrom AWRI 1176and research, toinvestigatethe potential to a financialsupporterofselectedInstitute yeastmanufacturermajor international and collaborative project withLallemandInc,a (ADWY). TheInstitutehasbeguna were availableasactivedriedwineyeast industry couldbeexpandediftheseyeast commercial quantities.Useofthestrainsby facilities topropagate theseyeastto requires experiencedstaff and suitable available asagarslopecultures, and S. bayanus S. cerevisiae. S. bayanus strains are only S. bayanus S. bayanus In barrel as providing in future yeastisolates. result inmore efficient characterisationof adjunct tophenotypicstudiesthat would of thestrains,thenitcouldrepresent an can beusedtopredict, inpart,thephenotype time- andlabour-intensive. IfAFLPanalysis new investigations willbeconductedonthese AFLP analysis.Future phenotypic the yeaststrainsappeartobeseparatedby different from asensorypointofview, and produced usingthesestrainscanbequite well characterisedattheInstitute.Wines strains, AWRI 1176and1375,havebeen This result isinteresting becausethe included intheanalysisforcomparison. of the however, ifthesestrainsare representative isolated attheInstitute.Ithasbeenunknown, of 1999 havedependedlargelyontwostrains Australian wineproducing regions since trialling bycommercial wineriesacross 707 and729).Theseprojects andextensive (Institute publications#632,647,679,693, sensory profiles ofwhiteandred wine to modulatethephysical,chemicaland potential fortheuseof recent yearshavedemonstratedthe Numerous studiesattheInstituteover winemaking strains Isolation ofSaccharomycesbayanus such as Other diversity ofpotentiallyusefulphenotypes. the remaining strains (Figure 5).Interestingly, 1439 and1443;oneincludingthe Sb3 andSb4;oneincluding1375,1438, groups: oneincludingthestrains1176, The strainsappeartoclusterintothree (AFLP) analysis(Institutepublication#589). Amplified FragmentLengthPolymorphism of thesestrainshasbeendeterminedby and JennyBellon,thegeneticrelatedness Dr MigueldeBarros Lopes,Jeff Eglinton molecular techniques.Incollaborationwith identity ofthestrainswasconfirmedby different regions ofAustralia.Thetaxonomic isolated from samplescollectedfrom been collected,withmoststrainsbeing Twelve strains of for wineproduction. and AWRI 796,remain widelyusedglobally #55 and156);strainsAWRI 350,AWRI 729 1960s and1970s(Institutepublications by DrBryceRankineandcolleaguesinthe landmark workcarriedoutattheInstitute species wasnotwellappreciated untilthe extent ofwinemakingproperties ofthat with can befound.We candrawananalogy isolates withbetterwinemakingproperties the genus S. bayanus. bayanus uvarum closely related tothetypestrainof S. bayanus S. bayanus S. bayanus S. cerevisiae Saccharomyces sensustricto S. bayanus cerevisiae ) thantheyare tothetypestrainof (a yeastthatisnowclassifiedas S. bayanus This observationsuggeststhat , AWRI 1176and1375, isolates appeartobemore isolates, butthisanalysisis S. bayanus strains, inwhichthe species, orifother and may harbourawide paradoxus, S. bayanus have now are species, strains S. S. the lipidcontent(determinedby level and absenceofalipidsupplement, since medium havebeenfermentedinthe presence grape juice.Thejuiceandmodel as farpractical,thecomposition ofthe grape juicemediumformulatedto mimic, profile determinedandachemicallydefined grape juicehasbeenselected,thechemical experimental conditions.AChardonnay establishing alaboratorymethodand Dr DiegoTorrea hascommencedstudieson assist winemakersin‘steering’winestyle. from chemicaldata.Suchknowledgewould to predict betterthesensoryprofile ofwine compounds whichcanultimatelybeused allow identificationofkeyimpactaroma quantitative descriptivesensoryanalysiswill aroma. Associationofchemicaldatawith conditions onthedevelopmentofwine of yeastspecies/strainandfermentation precisely establishtherelative importance Sefton’s teamprovides theopportunityto 30 yeastvolatilecompoundsbyDrMark isotope dilutionassaysforquantifyingsome The recent developmentofasuite stable rely heavilyonempirically-deriveddetail. these factors,andconsequentlywinemakers available ontherelative importance of published, littlesystematicinformationis Surprisingly, althoughmuchhasbeen temperature, agitation,solids,andso on. lipids, andphysico-chemicalconditions,pH, nutrients, sugar, aminoacids,oxygenand and strainofyeast,theconcentration volatile compounds,andincludethespecies to influencethequantitativeprofile ofyeast acids metabolism.Manyfactorsare known fatty acids,are derivedfrom sugarandamino comprise esters,alcohols,carbonylsand The majoryeastvolatilemetabolites,which referred toasthe‘fermentationbouquet’. the vinouscharacterofwine,andisoften products ofyeastmetabolismwhich provide responsible forvarietaldefinition;andthe secondary metaboliteswhichare largely on thebasisoftheirorigin:grapeberry can bedividedintoessentiallytwogroups The compoundsthatdefinewinearoma yeast derivedvolatilesofwine and fermentationconditionsonmodulatingthe Systematic studiestoestablishtheroleofyeast Saccharomyces Figure 5.Phylogenetictree (onestatisticallypossibleoutcome)showingtherelatedness ofdifferent Sb3, Sb4and1437-1444are isolatesof S. kudriavzevii ; BAY, species and S. bayanus 0.1 S. bayanus ; PAR, S. paradoxus isolates. Type strains:CER, S. bayanus ; CAR, . relative importanceoftheseconditions. style. Thisworkwillshedlightonthe will haveanimportantimpactonwine suggesting thatthechoiceofconditions compounds responds inadifferent manner fatty acids.Eachofthesegroups of aroma withrespectpattern toesters,alcoholsand a Chardonnay winehasrevealed acomplex oxygen andlipidsonthevolatilesprofile of thepresenceconcerning orabsenceof Preliminary analysisoftheresults and theformationofundesirablesulfide. effect onfermentationrate,residual sugar juices andmustshasaconsiderable found tobehighlyvariableinAustralian profile, sincethisnutrientsource hasbeen are importantindeterminingthearoma focus ondeterminingwhichaminoacids terms ofthevolatilesprofile, work will oxygen parametershavebeendefinedin published. Oncetheimportantlipidand impact onwinearoma profile hasbeen researchers butlittleinformationregarding fermentation hasbeenpromoted by some aroma profile. Theuseofoxygenduring yeast fermentationperformanceandwine another criticalyeastnutrientthataffects the presence andabsenceofoxygen, wine. Thefermentswere alsoconductedin that lipidsalsoaffect thearoma profile of performance andavailableevidencesuggests profoundly affect yeastfermentation of solidsingrapejuice)isknownto S. cariocanus S. cerevisiae ; UVA, S. uvarum ; MIK, S. mikatae . 1176,1375, ; KUD, CAR PAR 1444 1441 1442 1440 1437 1443 1438 1439 1375 Sb4 Sb3 1176 UVA BAY KUD MIK CER 21 Research Teams’ report 22 Research Teams’ report content ofaminoacidsandother investigate whetherchangesinthe current studyaimedtosystematically malolactic bacteria.Inthisregard, the on thegrowth andmetabolismof fermentation mayhavesignificant impact mannoproteins, duringandafteralcoholic compounds, includingpeptidesand complex yeast-derivednitrogenous amino acidsbyyeast,andrelease ofmore perspective, theuptakeandrelease of publication). From anutritional Research, Grape andWine Henschke 2003, malolactic bacteria(Gockowiakand metabolites thatare bioactivetowards nutrients, andtheproduction of to arisefrom competitionforessential of malolacticbacteria,andare considered shown toinhibitorstimulatethegrowth physiological interactionshavebeen submitted forpublication).Such Alexandre, Costello,etal.2003– of winemalolacticbacteria(reviewed by can haveonthegrowth andmetabolism the potentialinteractiveeffects wine yeast there isgrowing awareness of of wine, andnutritionalproperties physico-chemical white wines.Inadditiontoimportant wines andaproportion ofnon-aromatic process whichisappliedtomostred fermentation (MLF)remains anunreliable induction andcompletionofmalolactic selection andstarterculture preparation, Despite recent developmentsinstrain acids inaffectingcompatibility and malolacticbacteria–theroleofamino Physiological interactionbetweenwineyeast bacteria interactionsasitimpactsontheMLF. importance andnature ofyeastand N-heterocycles andestablishingthe bacterial originofthemousyoff-flavour Recent contributionshavecentred on the presence, inwine. compound, diacetyl,andcontrolling its role the oftheflavour defining ecologyofmalolacticbacteria,and the using moleculartoolsforunderstanding physiological compatibility, developingand on wineyeastandmalolacticbacteria Studies overthepastyearhavefocussed enhance commercial benefit. wine exploit MLFforguiding winemakers tomore efficiently control and outcomes ofthisresearch willallow secondary fermentationprocess. The the basisforunreliable inductionofthis wine. Asecondaryaimistoinvestigate modulating thesensorycharacteristicsof malolactic fermentationhason defining andunderstandingtherole that This project hasthelongtermaimof McCarthy, MelissaFettke Bartowsky, DrPeterCostello,Jane Staff: Malolactic fermentationandwineflavour Dr PaulHenschke,Eveline Australian Journalof accepted for style to negligible SO conditions (pH3.5,10%v/vethanol, were thentestedunderstandardised with regular stirring.Theresultant wines contact withyeastleesfortwoweeks immediately afterfermentation,or test winewere thenclarifiedeither were suppliedtoexcess.Sub-lotsofeach concentration, inparticularallaminoacids yeast nutrientsatnon-growth limiting was formulatedtocontainalltheprincipal Institute’s 2002AnnualReport.Themedium bacteria interactionsreported inthe the results ofanextensivesurveyyeast- wine yeaststrains,selectedonthebasisof commercial was fermentedtodrynesswiththree the studyofyeast-bacteriainteractions, juice medium,previously formulatedfor In thisstudy, achemically-definedgrape recently (Institutepublication#732). system.The methodologyhasbeenpublished bacteria inachemically-definedmodel thegrowthinfluence response ofmalolactic after extendedyeastleescontactcan and nutrients duringalcoholicfermentation autolytic release ofcellularproducts, and associated withayeaststrain-dependent in winemadebycontactwithyeast leesis is thatenhancedproliferation ofbacteria One possibleinterpretation oftheseresults peptides greatly increased. contact period,andtheconcentrationof to significantlydeclineduringthelees fermentation, yeastviabilitywasobserved production ofthelattertestwinesby amino acidcontent.Rather, during the was notassociatedwithanincrease in induced stimulationofbacterialgrowth had received yeastleescontact.Thislees- was greatly stimulatedbytestwinesthat strains, theoverallproliferation ofbacteria However, fortwoofthethree yeast depleted toagrowth-limiting concentration. would occurwhennonutrientshavebeen bacteria interaction,typicalofthatwhich corresponded toa‘compatible’yeast- bacterial growth response obtained chemically definedmedium.Thelevelof other nutrientsthatcomposethe mediated depletionofaminoacidsand/or generally notlimitedbytheyeast MLF infermentedtestwineswas results showthat bacterial growth and Under theconditionsofthisstudy, the five-week period. growth response wasmonitored overa strain-specific growth responses. Bacterial strains of were theninoculatedwithfourdifferent wine. Thetreated andreference testwines no supplementsservedasthereference thatreceived grape juicemedium.Wine selected from thechemically-defined with aminoacidsand/orothernutrients the wineswere treated bysupplementation nutrients neededforMLbacterialgrowth, whether theyeasthaddepletedessential growth response. Inorder todetermine Oenococcus oeni Saccharomyces cerevisiae 2 content) forbacterial to testfor during the2002vintagein Sauvignonwines conducted onCabernet (spontaneous) MLF, andfrom ayeasttrial from winesundergoinguninoculated Australian wineryisolateswere obtained are alsodepictedinFigure 6.The commercial cultures. These comparisons isolates andcompared thesewiththe similarity betweenAustralian In addition,wealsoexaminedthegenetic improved winemakingproperties. assist inthedevelopmentofstrainswith which, ifproved tobethecase,could RAPD markersandphysiologicalproperties, be neededtotestforassociationbetween wine conditions.Rigorous studieswould be especiallytolerantofseveraladverse most dissimilar, isclaimedbyLallemand to Interestingly, Lalvin41,whichisgenetically cultures are geneticallydistinct. shown inFigure 6,thefourcommercial examined usingtheRAPDtechnique.As amongst severalofthesecultures was European wineries.Geneticsimilarity wineries havebeenisolatedmainlyfrom in red andwhitewinesbyAustralian are widelyusedfortheinductionofMLF Commercial preparations of recently (Institute publication #733). and wineoriginhasbeenpublished differentiate methodology tounequivocally bacterium, successfully appliedtothemalolactic (Institute publication#718),was oxidative spoilageofbottledred wine associated withrandommicrobial Acetobacter pasteurianus (RAPD), whichwaspreviously applied to randomly amplifiedpolymorphicDNA Report. Thefingerprintingtechnique, summarised intheInstitute’s 2002Annual chain reaction (PCR)technology, as genome usingconventionalpolymerase generate asimpleDNAfingerprintofeach applied level,itisusuallysufficient to basis forstraindifferentiation. Atthe genetic similarityofstrains,andprovides a to determineintraspecificvariationor bacterial strainsprovides theinformation Comparative analysisofthegenome bacterium Oenococcusoeni commercial andwineisolatesofthemalolactic Genetic similarityandstraindifferentiationof Rosemount intheMcLaren Vale. undertook vintageexperiencewith Henschke supervisedthisproject. Melissa Costello, EvelineBartowskyandPaul first classhonoursdegree. DrsPeter Adelaide, forwhichshewasawarded a for herBAgScdegree atTheUniversityof Fettke attheInstituteduringhonoursstudies This workwasperformedbyMsMelissa testing toestablishitsvalidity. hypothesis, however, willrequire rigorous stimulate growth ofbacteria.Sucha offers thepossibilitythatpeptidesmight George Skouroumounis George Oenococcus oeni. O. oeni strains ofcommercial strains thatwere O. oeni O. oeni The RAPD that Hickinbotham Roseworthy Wine Science lees contact and timing of wine stabilisation common spoilage yeast. Readers are Laboratory (WSL). A preliminary analysis of with sulfur dioxide. A review article on referred to progress reports for projects the genetic relatedness of strains was given diacetyl and wine flavour has been prepared Evaluation of new analytical techniques in the Institute’s 2002 Annual Report. for publication, and signals the completion and of processing aids for winemaking and phase of this topic as a precursor study for Selection and improvement of wine yeasts The winery isolates are mostly genetically developing techniques to study the by application of molecular biology for distinct and the intraspecific variability relationship between bacterial metabolism additional reports on current progress. across Australian wineries is at least as and wine flavour. great as that represented by the four A review of the published literature on commercial European strains. Interestingly, Microbiological analysis of industry Dekkera/Brettanomyces shows that its several isolates from three wineries technical problems classical physiological properties of (Wineries I, V and IV, respectively) are relevance to alcoholic beverages (nutrient closely related to several commercial Staff: Dr Paul Henschke, Dr Eveline requirements, tolerance to pH, temperature, cultures (Lalvin EQ54, Viniflora oenos and Bartowsky, Dr Peter Costello, Jane alcohol, sulfite, etc.) have been, in broad Lalvin 31) suggesting that these isolates McCarthy, Peter Graves terms, described. However, little attention may be derived from commercial cultures, has been given to the strain variability of several of which have been used in This project undertakes to investigate specific these properties. The molecular characterisation Australia for five to ten years. The genetic winemaking microbiological problems of of Australian isolates in progress at the variability of O. oeni in Australian wineries importance to the wider industry. Recent Institute (see progress report for project suggests that these indigenous strains topics have included mousy-off flavour Selection and improvement of wine yeasts possess a wide range of physiological and occurrence and formation in wine, by application of molecular biology) biochemical properties which could be Dekkera/Brettanomyces spoilage of wine, suggests a wide genetic diversity of strains, expected to reflect the wide diversity of and wine microbiological spoilage and therefore the possibility of high wine environments. workshop as summarised in the previous variability in physiological and biochemical annual report. Work has continued on the properties of relevance to wine production. Malolactic fermentation and wine flavour – Dekkera/Brettanomyces spoilage project Therefore, our focus will be on the diacetyl and the ‘buttery’ attribute, a and on a case study concerning microbial characterisation of Australian isolates as precursor investigation mediated oxidative spoilage of bottled red opposed to culture collection strains that wine. Small studies concerning application form the basis of much of the published One of the main volatile flavour components of lysozyme in winemaking, yeast spoilage work. For example, strain variability with attributed to MLF activity is diacetyl (2, 3- of sweet wine and alcohol yield of respect to important wine parameters butandione). Diacetyl confers a ‘buttery’ attribute to the wine, which, depending Table 3. Suggested guidelines to manipulate the diacetyl content and ‘buttery’ flavour of wine using malolactic fermentation upon the concentration and wine style, is perceived as desirable or an off-flavour. Through a survey of 93 commercial bottled Diacetyl content Winemaking manipulation wines, we have recently shown that the perception of diacetyl is not only dependent High Bacterial strain with high potential for diacetyl production upon its concentration, but also on the Lower inoculation level presence of other wine aromatic components, Increase redox potential including fruitiness, aged aromas and Avoid contact with yeast lees dominant oak-like aromas (Institute Stabilise wine immediately after malic and citric acid degradation publications #689 and 709). Figure 7 illustrates this point for Australian Low Bacterial strain with low potential for diacetyl production Chardonnay and Shiraz wines by Higher inoculation level comparing diacetyl concentration with the Prolonged contact with yeast and bacteria lees before stabilising wine ‘buttery’ aroma score as determined by a trained sensory panel. In this survey, the diacetyl concentration was close to its fermentation yeast were also undertaken. seems to be underestimated. In common published sensory threshold value for Project AWR 5 also provides resources for with Saccharomyces yeast, Dekkera/ Chardonnay wines and below the sensory maintaining the Institute's culture collection Brettanomyces yeast appear to have very threshold value for Shiraz wines, which provides yeast and bacteria for simple nutrient requirements suggesting that suggesting that Australian winemakers teaching and research, performing this feature could, under some conditions, currently favour a wine style having a low advanced wine microbiological analysis, offer a means for control. For example, it has ‘buttery’ impact. Diacetyl concentration undertaking dried yeast quality testing, and been established by Chatonnet’s group in was also observed to decline with bottle providing microbiological consultation to Bordeaux, that lowered residual sugar content age for the set of red wines studied, which industry personnel. of wine can be used to limit growth and would further reduce the ‘buttery’ character. volatile phenol formation. Comparative Dekkera/Brettanomyces – Isolation from wine physiological characterisation of Australian Because of the biological origin and and a preliminary physiological characterisation isolates can be expected to point to chemically reactive nature of diacetyl, additional control measures. especially its ability to form an adduct with As forecast in the Institute’s 2002 Annual sulfite and its susceptibility to bio-reduction Report, several of the aims of the Institute’s The origin of Dekkera/Brettanomyces yeast in the to the diol, there are numerous winemaking multidisciplinary collaborative Dekkera/ winery is still unclear, and may include infected practices that can be used to modulate the Brettanomyces project, coordinated by Peter wine, poor sanitation of equipment and the desired concentration and sensory impact Godden through the GWRDC-funded common fruit fly. A recent publication (Connell et of diacetyl in wine. These practices, which project Evaluation of new analytical al. 2002 AJEV 53:322-324; 2002) suggests that have been published (Institute publications techniques and of processing aids for this yeast can be spread through a winery by air #637 and 689) are summarised in Table 3. winemaking, was to acquire a knowledge currents. The physical environment of a winery Important factors include bacterial strain, base of published work, isolate and may also prove to be a risk factor in inoculation rate, wine chemical composition characterise strains from Australian wines, Dekkera/Brettanomyces contamination and (in particular, concentration of citric acid and ultimately to provide information to control. The molecular characterisation of which is the bacterial substrate for diacetyl), winemakers for practical control of this Australian isolates in progress at the Institute will report Teams’ Research 23 24 Research Teams’ report .005 .008 1.00 0.85 0.70 0.55 0.40 The panelwasscreened fortheirability todetectdiacetylinwineandwere involvedinnumerous training sessionswhere their marking adiacetylconcentrationof0.2mg/L for Chardonnay and2.8mg/LforShirazwines indicatetherespective reported sens Figure 7.Thediacetylconcentration and‘buttery’sensoryperception of36AustralianbottledChardonnay and29Shirazwines. ScienceLaboratory(WSL)during2002vintage. yeast trialconductedintheHickinbothamRoseworthyWine Figure 6.Clusteranalysisof

Diacetyl (mg/L) 0 1 2 3 0 1 2 3 eoocsoeni Oenococcus strains isolatedfrom different Australianwineries(ItoVII),commercial cultures Sauvignon andaCabernet Chardonnay Shiraz performancewasassessed. Thehorizontallines ory thresholds fordiacetyl. 0 1 2 3 4 5 0 1 2 3 4 5

‘buttery’ aroma score WSL rotary WSL rotary WSL rotary WSL rotary Lalvin 41 V Winery II Winery IV Winery III Winery WSL static12 WSL static11 IV Winery VI Winery VII Winery WSL rotary WSL rotary WSL rotary WSL static7 IV Winery WInery IV Lalvin 31 WSL static10 WSL static9 WSL static8 V Winery oenes Viniflora I Winery Lalvin EQ54 also contribute to this question (see progress Microbial spoilage of bottled red wine – preliminary cork closure. This hypothesis, which still report for project Selection and improvement of evidence for the role of the cork closure requires careful experimental verification, wine yeasts by application of molecular biology). has been recently published (Institute In collaboration with a large wine company publication #718). Dekkera/Brettanomyces yeast generally proliferate we previously investigated the microbiology only slowly in wine and cell numbers are often of a series of batches of red wine which had Although the incidence of this type of wine very low, necessitating efficient recovery. become oxidatively spoiled post bottling spoilage is unknown and likely to be relatively Classical methods rely on the use of elective (Technical Review issue #127). The bottles low, a second occurrence of a large batch media to target Dekkera/Brettanomyces yeast had been stored in an upright position for of bottled Shiraz wine recently has provided followed by simple test procedures that provide various periods of time within a year as us with the opportunity to test our hypothesis. a presumptive identification. They have opposed to storage in a horizontal position The spoiled wines show similar characteristics morphological, physiological and biochemical or being sealed with a ROTE closure. A low to those of the previous study, and again a characteristics that assist their selective isolation proportion of bottles, at random, in various closely related group of acetic acid bacteria from wine: ogival and profusely elongate cells, wine batches exhibited various degrees of were found. The degree of visible spoilage slow colony growth, acid production, tolerance oxidative spoilage, ranging from loss of varied amongst bottles and examples are to cycloheximide and growth on L-lysine and fruit to overt oxidation (acetaldehyde, ethyl shown in Figure 8. Measurement of the ethanol. A survey of various media has confirmed acetate and acetic acid), which was dissolved oxygen concentration of visibly that the recently developed Differential Dekkera/ confirmed by chemical analysis of spoiled spoiled and unspoiled bottled wines showed Brettanomyces Medium published by Louriero and non-spoiled wines. a difference of up to ten-fold between the and colleagues (J. Appl. Microbiol. 90:588-599; two sets of wines. Sensory analysis of the 2001) is more effective than conventional media Spoiled bottles were identified by a wines correlated well with the higher supplemented with the fungicide cycloheximide, characteristic deposit on the inner surface acetaldehyde, ethyl acetate and acetic acid although this medium still supports the growth of the neck of upright stored bottles, concentrations of severely spoiled wines. A of several other yeast species. Identification is giving a ring-like appearance when the small but significant decrease in alcohol confirmed by a DNA (RFLP) method (see progress bottle was tilted (Figure 8). Bacteria could content was also noted in heavily spoiled report for project Selection and improvement of be isolated from both spoiled and non- wines, consistent with the oxidation of wine yeasts by application of molecular biology). spoiled wines, and were found to belong ethanol. The presence of bacteria and to a closely related group of Acetobacter concentrations of acetaldehyde, ethyl acetate A related issue seems to be the difficulty of pasteurianus strains, based on molecular and acetic acid correlated well with the recovering Dekkera/Brettanomyces from some identification techniques developed for the degree of wine spoilage, as noted by the wine samples despite high 4-ethylphenol project (Institute publication #718). However, thickness of the ring-shaped deposit on the concentrations. The Industry Services Laboratory oxidative metabolism of ethanol only neck of the bottle. has identified a number of such wines during occurred in a small proportion of bottles, their survey of Australian wines obtained as apparently distributed at random within a This study further links the natural variability bottles and barrel samples. The phenomenon bottling run. As similar bacteria were of cork closures to oxygen permeability to ‘viable but non-culturable state’ is a well-known present in spoiled and non-spoiled bottles, differential changes in wine composition, problem in the food industry whereby processed we hypothesize that the upright bottle at least in conjunction with prolonged foods are often spoiled by ‘cryptic’ spoilage storage position created a heterogeneous upright storage of bottles. Direct microorganisms which are difficult to cultivate. environment which allowed the growth of measurement of oxygen permeation by the We encountered a situation with this bacteria in only those bottles sealed with cork closures used in this trial could be characteristic when attempting to isolate acetic cork closures that had an upper limit for expected to strengthen our hypothesis. acid bacteria from bottled red wine (Institute the natural permeability to oxygen. Such These two unconnected cases of bottled publication #718). To test for this phenomenon, an heterogeneous environment would not wine spoilage highlight the importance of Dr Peter Costello, assisted by Peter Graves, exist in horizontally stored bottles since the good winery practice with respect to the commenced a series of tests designed to larger volume of wine adjacent to the cork preparation of wine for bottling and the resuscitate viable but non-culturable Dekkera/ would strongly compete with the bacteria minimisation of storage time of bottled red Brettanomyces yeast. To date, we have evidence for the oxygen as it diffuses through the wine in a vertical upright position. that this is the case for the one wine studied which contains over 800 ppb 4-ethylphenol. A genetic fingerprint of the isolates will be made to negate the possibility that a variant form or a different strain/species of yeast has been recovered. These are early indications and need to be verified for other wine samples.

none fine

medium heavy

Figure 8. Visible deposit of acetic acid bacteria on the neck of bottles of spoiled red wine held in a tilted position Research Teams’ report Teams’ Research 25 Lysozyme and winemaking S. cerevisiae strains for effect on wine colour capacity rotary fermentors. Alcoholic and density and phenolic concentration of a malolactic fermentations proceeded Lysozyme, a protein that is present in various young Shiraz wine was summarised in the efficiently with each of the three yeast strains. bodily fluids and that is an important previous annual report. Analysis of the results The Shiraz wines are currently being bottled component of the eukaryotic immune system, show that the yeast strains could be divided for sensory analysis later this year. From initial is a natural bacteriolytic enzyme. It has into three statistically significant groups, that colour and phenolic analysis, the low and several potential beneficial applications in is, yeast that gave low, medium or high wine high colour yeast strains produced wines wine production for which the Australian colour density. We further confirmed these that were significantly different in wine wine industry is applying to permit its use. results with Shiraz fruit obtained from other colour density and pigmented polymers For example, lysozyme can be used to inhibit viticultural regions, and with frozen versus (Figure 9), corroborating our earlier findings certain bacterial species or delay the onset fresh fruit. These results suggested that there with the micro-vinification system. of malolactic fermentation, lowering the are intrinsic differences between some yeast The success of the pilot scale vinfication trials

need for SO2 addition in such applications. Lysozyme is only effective against Gram Table 4. Factors affecting lysozyme activity and wine stability positive bacteria (including lactic acid bacteria) and not Gram negative bacteria Wine variable Effect on lysozyme activity or wine stability (including acetic acid bacteria, such as Acetobacter) or yeast, such as Saccharomyces and Dekkera/Brettanomyces. Our survey of Wine type Effective in both red and white wines the susceptibility of wine related species of Species/strain of lactic acid bacteria Variation in susceptibility between genera, malolactic bacteria, Lactobacillus (three species and strains of bacteria species) and Pediococcus (three species) Lactic acid bacteria initial concentration Effective against most species and strains and four strains of O. oeni to lysozyme in of lactic acid bacteria up to 106 cfu/mL white and red wine did, however, indicate Wine pH Activity increases as the wine pH increases significant intragenic variability, with O. oeni Wine alcohol content No influence on lysozyme activity strains being especially susceptible. Phenolic compounds Wine phenolics may precipitate with lysozyme and decrease its activity Research conducted in collaboration with Fordras A/S, a European manufacturer of Protein stability (haze) May induce haze formation in white wines lysozyme, has highlighted the potential use Bentonite Decrease in antimicrobial effect if used in of lysozyme to control bacterial growth in conjunction with bentonite both white and red wine, and also identified Other fining agents These can bind lysozyme and reduce its activity areas where caution is needed. The (carbon, silica sol, oak chips and tannin) antibacterial action of lysozyme is relatively Sparkling wine foaming properties In some cases may slightly increase foam stability high at wine pH (~3-4) and unlike sulfur Other wine additives A heavy haze may form when used in dioxide, lysozyme is also effective at higher conjunction with metatartaric acid wine pH. The use of lysozyme in winemaking Organoleptic properties of the wine Minimal impact on aroma and palate has can only replace the antimicrobial activity been reported and not the antioxidant activity of sulfur dioxide. The factors that affect lysozyme activity in wine, highlighted in Table 4 and strains, confirming the observations by conducted in the WSL has depended on recently published (Institute publication various winemakers and Lallemand. several important factors. First, enormous #728), include initial bacterial cell numbers, commitment and cooperation of the species and strain, wine pH, phenolics content Comparison of the micro-vinification collaborating winery not only for providing and the use of bentonite. Our research has methology, based on 1 kg of fruit, with a large quantity of fruit to the Institute’s demonstrated that there is minimal impact pilot scale winery fermentation using 600 kg specification, but also for organising grape of lysozyme on the sensory properties of fruit has shown that under certain conditions harvest such that 10 tonnes of grapes was both red and white wine, an observation wines of comparable composition can be divided across multiple grape bins to also noted by other research groups. obtained, as given in the previous annual achieve a more uniform distribution of report. The trial was conducted with 2002 must composition in the winery. The Hardy Yeast interaction with grape phenolics and effect Cabernet Sauvignon fruit harvested from Wine Company Management (specifically, on wine sensory properties the Orlando Wyndham Padthaway vineyard Peter Dawson and Angus Kennedy) is in the Hickinbotham Roseworthy Wine gratefully acknowledged for generously Staff: Dr Eveline Bartowsky, Simon Dillon, Science Laboratory (WSL) in conjunction providing the fruit as are Ben Vagnarelli and Professor Peter Høj, Dr Paul Henschke with the Institute’s Tannin team. Mark Zeppel who organised the logistics of Nevertheless, it is necessary to confirm the fruit harvest and delivery. The University of A collaboration between the Institute and magnitude of effect that yeast strains have Adelaide’s winemaker Stephen Clarke and Lallemand has continued to investigate the on colour density and phenolic composition winery assistant Chris Day are thanked for interactions between yeast and grape phenolics when conducted on a winery scale, and processing the must and producing bottled during red wine fermentations. Lallemand especially to determine the sensory wine to specification in the WSL. Close is contributing a substantial sum of money properties of wines made with these yeast cooperation of the AWRI-Lallemand project for our investigations, for which we are most under controlled conditions. and Wine Microbiology teams, Simon Dillon, appreciative. The donation of Shiraz grapes Eveline Bartowsky, Jane McCarthy and Paul by the Hardy Wine Company and Orlando This vintage, a pilot-scale vinification trial was Henschke, and the Institute’s Tannin, NIR Wyndham for these studies is also appreciated. again conducted in the WSL under the and Sensory teams, Markus Herderich, Leigh supervision of winemaker Stephen Clarke Francis, Mango Parker, Mariola Kwiatkowski, The background to this project, development with assistance from Chris Day in conjunction Daniel Cozzolino, Bob Dambergs and Kate of a micro-vinification methodology and with the Tannin team supervised by Dr Markus Lattey was essential. This trial has again comparison of its performance with pilot Herderich. Shiraz grapes from Banrock highlighted the need for careful planning scale winery fermentation, and survey of Station, Berri/Riverland, were generously and a large team of highly motivated commercial Lallemand yeast strains for impact provided by the Hardy Wine Company. individuals in order to successfully on wine colour and phenolics content has Three yeast strains, identified from previous undertake a large-scale project. been described in previous annual reports. trials, were used to produce the wines with 26 report Teams’ Research The findings of a survey of 17 Lallemand replicated batches of must processed in 900L genes forwineyeastimprovement. More flavours towine,andtheidentificationof interspecific hybridsthatimpartdiverse low ethanolwineyeast,theproduction of from previous years:thegenerationofa Research iscontinuingonthree projects wines withimproved sensoryqualities. emphasis ongeneratingyeastthatproduce providing improved winestrains,withan The molecularbiologyteamisfocusingon Analysis Facility Derek vanDyk,AustralianProteome Carolyn Leach,UniversityofAdelaide,Dr Jelle Lanstein,UniversityofAdelaide,Dr Vladimir Jiranek,UniversityofAdelaide,Mr Fleet, UniversityofNewSouthWales, Dr University ofTechnology, Professor Graham Collaborators: Paul Henschke,SakkiePretorius, PeterHøj Pollnitz, Tracey Seibert,MarkSefton, Daniel Cozzolino,Gordon Elsey, Alan Eveline Bartowsky, DimitraCapone, Guzinski, AnthonyHeinrich,KateHowell, Jeff Eglinton,Lorelie Flood,Jaromir Staff: application ofmolecularbiology Selection andimprovementofwineyeastsby Peak area units Absorbance units 1000 1500 2000 2500 500 Miguel deBarros Lopes,JennyBellon, 5.0 5.5 6.0 6.5 7.0 7.5 8.0 0 with low, medium andhighcolourdensity Saccharomyces cerevisiae Shiraz grapemust(Berri/Riverland,2003)byfermentation in900Lcapacityrotary fermentorswith Figure 9.Comparisonofwinecolour densityand‘pigmentedpolymers’ofyoungwinesmadefrom Dr PaulChambers,Victoria low low strains previously identifiedthrough small-scalefermentationstogivewine Wine colourdensity Wine Pigmented polymer eimhigh medium eimhigh medium dramatic effect onthebiosynthesis of a metabolism hasbeenshowntohavea Altering theyeast’s glycerol andaceticacid for reducing aceticacidtoacceptablelevels. dehydrogenase, provided aneffective remedy ( in wine.Thedeletionofasecondgene concentration thatisgenerallyunacceptable acid concentrationinwineabove1g/L,a strains, however, alsoincrease theacetic strain duringfermentation.Thesemodified produce lessethanolthananon-modified gene ( Strains overexpressing theglycerol synthesis Construction ofawinestrainproducinglessethanol of Adelaide. with DrVladimirJiranekfrom TheUniversity Productive collaborationsare alsocontinuing South Wales andDrEvelineBartowsky. Graham Fleetfrom TheUniversityofNew Kate Howellisco-supervisedbyProfessor South Wales whorecently joinedtheteam. PhD studentfrom TheUniversityofNew yeast isbeingperformedbyKateHowell,a the mechanismofvolatilethiolrelease by the Institute.Anewproject, tounderstand can befoundinprevious AnnualReportsof background informationontheseprojects ald6 ∆ GPD2 ), whichencodesanaldehyde ) accumulatemore glycerol and cerevisiae commercial wineyeast( Interspecific non-GMOhybridsbetween Hybrid wineyeasts these mutants. and whatothermetabolicchangesoccurin strains are lowethanolproducing strains It needstobedeterminedwhetherthese improved growth underselectiveconditions. have beenisolated,andthesemutantsshow commercial wineyeaststrainswithmutations be usedimmediatelybytheindustry. Two low ethanolwinestrains,whichcanthen now takinganon-GMapproach togenerate volatile acidity. Lorelie Floodis,therefore, necessarily havingaparallelincrease in at theexpenseofethanolproduction without of carbohydratetowards glycerol production yeast are abletodivertconsiderableamounts a moleculargeneticapproach indicatesthat The informationgainedabovebymeansof (Institute publication#684). number ofotherpotentialflavourmetabolites analysed inthesewines. aroma compoundsare currently being wine yeast.Thequantificationofkey ferments asefficiently asthecommercial results indicatethatthehybridyeast properties. Preliminary winefermentation appears tohaveinteresting organoleptic less robust thancommercial winestrains, yeast hasbeenwellstudied,andalthough winemaking properties ofthe AWRI 1375hasbeensuccessful.The commercial wineyeastand The generationofhybridsbetweena with novelwinemakingcharacteristics. an effective strategyforacquiringyeasts demonstrate thatinterspecifichybridsoffer to oneoftheparents. Theseresults concentration oracomparable showed eitheranintermediate than thewineyeast.Thehybrid respectively inthe much lower(onesixth)concentration produced atamuchhigher(20-fold) anda 2-phenyl ethylacetate(honeyaroma) are compound withablackcurrantaroma, and example, dihydro-2-methyl-thiophenone, a ferments ofthetwoparental strains.For a largevariationinconcentrationbetween these twostrains.Somemetabolitesshowed S. paradoxus commercial wineyeast(AWRI 838),a flavour metabolitesinfermentsmadebya the concentrationsofseveralpotential Jeff EglintonandAlanPollnitzhave compared chromatography/mass spectrometry (GC/MS) considered tobemore complex.Usinggas hybrid winesbeinggenerallypreferred and using acommercial wineyeast,with the were notablydifferent tothewinesmade demonstrates thatthehybrid-madewines and commercial trials.Sensoryanalysis juice efficiently inbothsmall-scalewinemaking strains havebeenshowntofermentgrape (Institute publication#685).Thesehybrid species havebeenmadebyJennyBellon ) andotherrelated yeast orahybridbetween S. paradoxus Saccharomyces S. bayanus Saccharomyces S. bayanus parent strain 27 Research Teams’ report 28 Research Teams’ report 160 240 320 400 480 560 640 720 800 880 960 laboratory strainsismarkedlydifferent. that theprotein expression ofwineand electrophoresis hasbeenusedtodemonstrate generally unknown.Two dimensionalgel the desirableattributesofwinestrains are The genesandproteins thatcontributeto consistent withwinearoma andflavour. for thesynthesisofmetabolitesthatare commercial winestrainshavebeenchosen fermentation respectively. Furthermore, and ethanolatthebeginningendof including thehighconcentrationsofsugar a numberofstresses, to withstand and wineyeastneedtobeable environment strains. Thewinehabitatisademanding differences whencompared tolaboratory wine strainshaveimportantphysiological Although representing thesamespecies, knowledge onitsbiologyisavailable. for thisreason anextensiveamount of model organismforscientificresearch, and Saccharomyces cerevisiae Identifying genesforwineyeastimprovement S. kudriavzevii commercial wineyeastpossessing hybrid wineyeast.Thisisthefirsttimea S. cerevisiaeXSaccharomyceskudriavzevii and JennyBellonhaveidentifieda studies performedbyAnthonyHeinrich and yeast S6Uisahybridbetween demonstrated thatthecommercial wine length polymorphism(AFLP)have previous studiesusingamplifiedfragment Natural interspecifichybridsalsoexist.Our be enhanced. flavours ofthenon- genome sothatthenovelaromas and may offer ameanstoalterthehybrid strain. Theseresults are importantasthey more efficiently thantheparental hybrid the parents. Thishybridappearstoferment that haslostachromosome from oneof Genetics, ischaracterisingahybridisolate The UniversityofAdelaide,Department An honoursstudent,Jaromir Guzinski,from 80 0 S. bayanus 3 4 5 6 170 160 150 140 130 combination of the the of combination Figure 10.AFLPofhybridwineyeast.Fingerprintthecommercial wine DNA hasbeenidentified. (Figure 10).RecentAFLP S. cerevisiae S. cerevisiae is animportant S. cerevisiae (shown inblue)andthe parent can when cellsrespond tohighconcentrations is onlyexpressed inalaboratorystrain although highlyexpressed inawineyeast, metabolism andathird protein that, protein, aprotein involvedinonecarbon have beenchosenforfurtherstudy, aglycolytic proteins havebeenidentified,three proteins identified usingMS/MS.Althoughmany and presently otherproteins are being with Derek vanDykatAPAF iscontinuing, proteins havebeenidentified.Acollaboration spectrometry (MALDI-TOF)anumber of laser ionisation/desorptiontime-of-flight University inSydney. Usingmatrix-assisted Analysis Facility(APAF), locatedatMacquarie spent sixweeksattheAustralianProteomics student from TheUniversityofAdelaide, this research AnthonyHeinrich,aPhD for and interest istechnicallydemanding fermentation. Identifyingproteins of relation toissuessuchasstuck performance canbefurtherenhancedin wine strainsmightthuspointtohowstrain quantity betweenlaboratorystrainsand Insights intowhatproteins are differing in S. bayanus

Sakkie Pretorius (shown ingreen) fingerprint. yeast S6U(showninred) isa

Paul Chambers Anthony hasbeeninvaluable. data obtainedlaboratory totheproteome by the transcriptomedata from yeast acquire ethanoltolerance.Comparing been interested inunderstandinghow last fewyears,Paulandhiscolleagueshave laboratory foranextendedstay. Overthe of Technology visitingthe inMelbourne Paul Chambersfrom University TheVictoria We havealsobeenfortunatetoDr proteins supporttheprediction. strains lackingthedifferentially expressed Cozzolino andtheNIRteam,indicatethat Spectroscopy (NIR),performedwithDaniel Preliminary results usingNearInfra-Red contribute towineflavourandaroma. during fermentationandbydoingso, metabolites produced bythewineyeast protein expression mayalsoaffect the It isexpectedthattheobservedchangesin and Western analysis. PCR are beinganalysedusingRealTime these proteins theexpression ofthesegenes of ethanol.To further explore therole of Paul’s

Miguel de Barros Lopes Volatile thiol formation by yeast with a model protein, BSA, as part of the during fermentation C method optimization experiments, has suggested that absorption is pseudo The important role of yeast in modifying the irreversible. This supports work by others chemical, mouth-feel and flavour complexity demonstrating that removal of absorbed of wine is gaining increased recognition. A protein from bentonite lees is difficult class of yeast metabolites that have an and that bentonite regeneration is not a important influence on wine aroma and 4mmp conc. BDsimple process. A flavour are the sulfur containing volatile thiols. These compounds have been found in a We plan to study the absorption onto number of red and white wines, and have bentonite of the two main unstable wine been shown to contribute significantly to Figure 11. 4MMP release by different proteins, grape thaumatin-like proteins and varietal aromas of Sauvignon Blanc wines. S. cerevisiae strains chitinases. These proteins are not One of the most potent thiol compounds is commercially available so purification of 4-mercapto-4-methylpentan-2-one (4MMP), Studies on unstable wine proteins involved in these proteins from a Muscat Gordo juice which has a perception threshold of 0.1 ng/L haze formation concentrate has commenced. and 3 ng/L in water and wine respectively. Depending on the concentration, this Staff: Dr Elizabeth Waters, Dr Miguel de Barros Use of haze protective mannoproteins as an compound has an aroma of blackcurrant or Lopes, Professor Peter Høj, Ken Pocock, alternative to bentonite fining cat urine/box tree. 3-Mercaptohexan-1-ol Shauna Brown, Yoji Hayasaka, Gayle (3MH), having a passion-fruit aroma, is Baldock, Richard Muhlack, Jean McIntyre The putative haze protective mannoprotein another important thiol that has been found genes from yeast (HPF1, HPF1’ and HPF2) at concentrations approximately 100-fold Collaborators: Dr Filomena Pettollino (CRC have been cloned and all three have been higher than its threshold concentration. for Bioproducts), Dr Chris Colby, Dr Brian successfully deleted individually and in O’Neil (School of Chemical Engineering, combination in haploid and diploid laboratory The thiols are predominantly, if not exclusively, The University of Adelaide), Audrey Lim yeast strains. Analysis of the haze protective present in grape juice as non-volatile cysteine (Hardy Wine Company), Dr Eileen Scott, Dr activity of the material from over expression conjugates. Predicting the concentration of Belinda Stummer (School of Agriculture and deletion strains has supported our the flavour precursors in grapes is not possible and Wine, The University of Adelaide), identification of these genes as coding for at present. However, in Sauvignon Blanc grapes Natalie Fryar (Yalumba Wines) haze protective factors. By using a it has been reported that the concentration hexahistidine labelled form of Hpf2p (6xhis of 3MH precursor is approximately 10 times Visual clarity is important to white wine Hpf2p) we have also obtained evidence higher in the grape skin than in the juice. quality. Sediments and hazes can form after that we have successfully identified a gene Taking into account the respective proportions bottling due to the denaturation of grape coding for a haze protective factor. of juice and skin, this means that approximately protein. Winemakers usually remove protein 60% of the compound is in the skin. The from wine by fining with bentonite clay Phenotypic analyses have suggested that 4MMP precursor is found mostly in the juice and although this process is effective it has the haze protective mannoproteins may have as it is present in similar concentrations in several drawbacks, including losses of wine some impact on the ability of yeast to tolerate skin and juice. The conversion of the non- in bentonite sediment, problems of disposal cold and ethanol. This is an unexpected volatile grape precursors to volatile thiols of bentonite waste, and possible effects on bonus from the HPF project and illustrates requires enzymic action. Although the wine flavour. Work on improving bentonite the serendipitous nature of research. extraction of the cysteine conjugated efficiency has, therefore, been initiated this Further investigation of the deletion strains precursors into the juice appears to be year. In addition, we have continued work has suggested that Hpf1p and Hpf1’p are correlated to the final concentrations of the on the use of haze protective mannoproteins also involved in yeast mating. volatile thiols present in wine, only a small and from yeasts as alternatives to bentonite varying proportion of the precursor is converted fining. Further progress has also been made A heat unstable Sauvignon Blanc wine to the active aroma compound during on putting the ‘nuisance’ grape proteins to containing a hexahistidine labelled form of fermentation. This conversion appears to be work as markers of grape variety. Hpf2p (6xhis Hpf2p, 200 mg/L), invertase yeast strain dependent. (200 mg/L) or no addition (‘control’) has been Improving the efficiency of bentonite fining prepared and stored at 25˚C. After three A project has been initiated to provide through chemical engineering and six months’ storage, both 6xhis Hpf2p winemakers with the ability to regulate the and invertase appeared to be as active as amounts of the volatile thiols released As the first step in improving bentonite they were when the trial was set up, indicating during fermentation. Together with Gordon efficiency, studies with our collaborators at that the short term stability of these haze Elsey, Mark Sefton and Tracey Siebert, the School of Chemical Engineering, have protective mannoproteins in wine is good, sensitive methods using the GC/AED (Gas been undertaken on how proteins absorb even at slightly elevated temperatures. Chromatography/Atomic Emission Detector) onto bentonite. Data from initial studies have been developed to accurately measure the concentrations of 4MMP and 3MH in model fermentations. With these methods, it has been shown that commercial strains differ greatly in their ability to release 4MMP (Figure 11), confirming observations of Dubordieu et al. in Bordeaux. The mechanisms of release of the volatile thiols from the cysteine bound conjugates are being studied. It is expected that this will provide winemakers with the knowledge to select specific commercial yeast strains and to use various winemaking practices that will produce wines with the desired concentration of these important aroma compounds. Research Teams’ report Teams’ Research 29 30 Research Teams’ report was furtherconfirmed thisseason.Five the varietyofgrapesfrom analysingwine The abilityofthemethodtodifferentiate Use ofmassspectrometrytodifferentiate varieties protective material. allow non-GMOproduction ofhaze is responsible forhazeprotection. Thiscould works andwhichpartoftheHpf2pmolecule techniques. Theaimsare tofindouthowit chemistries, enzymicdigestion,etc.) and biological(protein andcarbohydrate microscopy, GPCMALLS,etc),chemical physical (laserlightscattering,atomicforce to studythemechanismofactionusing team atCRCforBioproducts are continuing which thisoccursisnotunderstoodbutthe and finalparticlesize.Themechanismby haziness byeffecting protein aggregation protective mannoproteins, reduces visible and indicatethatHpf2p,likeotherhaze factors (Institutepublications#439,639) with lesspure samplesofhazeprotective trends are similartothoseobservedpreviously when compared withthecontrol. These concentration bothbefore andafter heating in particlesizewithincreasing 6xhis Hpf2p Hpf2p showedthatthere wasadecrease and heatedwinesamplescontaining6xhis Analysis ofthesizeparticlesinunheated The mechanismofhazeprotectiveactivity on theirspecifications. individual companiesandwoulddepend that needstobemadeaftertrialsby forcommercial release isadecision enough stability wouldbeconsideredheat stable yeast. Whetherthiswine,withimproved release ofhazeprotective factorsby fermentation maybeduetoadditional heat stabilityofwinesfollowingsecondary These results suggestthatimprovement in tirage thanatthebeginningoftrial. appeared tobegreater atfourweekspost protective mannoproteins inthiswine proteins wasunchanged.Thelevelsofhaze though theamountofunstablewine of thiswineimproved followingtirageeven the experiment.However, theheatstability basis ofan80ºCsixhourheattest,throughout and thislotremained heatunstable,onthe was notstableduringsecondaryfermentation, fermentation andageing.Theunfinedlot bottled lotsremained soduringsecondary bentonite were heatstable,andthese The basewinestreated withbothratesof aged onleesundercommercial conditions. put through secondaryfermentationand different levelsofbentonite.Theselotswere base winewastreated witheithernoneortwo toexplore thisphenomenon.One Wines, collaboration withNatalieFryar, Yalumba A trialwasinitiatedinDecember2001 immediately followingsecondaryfermentation. protein unstablebasewinesbecomestable Industry practitionershaveobservedthat secondary fermentation Haze protectivemannoproteinsduring grapes toeffect theabilityof theelectrospray reason forpowderymildewinfectionof maintained. We, therefore, currently seeno characteristic qualitativeprotein profile was powdery mildewinfectionofgrapes, the in protein levelsinjuiceandwinedueto a heattest.Despitethequantitative changes the levelofhazinessinwinefollowing infected), thishasasignificantimpacton levels ofinfection(>30%bunches in Chardonnay juiceandwine.Athigher levels ofonethethaumatin-likeproteins powdery mildewinfectionincreases the CRCV Project 1.5.2,hasshownthat thr TheUniversityofAdelaide Wine, Stummer attheSchoolofAgriculture and in collaborationwithEileenScottandBelinda Teresa Girbau,andfollowed upthisvintage student from theUniversityofBarcelona, Work performedlastvintagebyavisiting publications #663and734). varieties inawine(seealsoInstitute used toexcludethepresence ofcertain for varietaldifferentiation butcouldbe proteins aloneare probably notsufficient differences intheMrofthaumatin-like variable group ofwineproteins. The levels ofchitinases,themostvarietally proteins butdonotcontaindetectable detectable levelsofthethaumatin-like stabilised withbentonitecontainlowbut challenge tothismethod.Whitewines Bentonite finingpresents aparticular the varietalidentification. spectrometry withoutpriorknowledgeof for allofthenon-finedwhitewinesbymass the grapevarietyforMerlotwineand spectrometry operatortocorrectly identify of protein, itwaspossibleforthemass from red grapes.Despitethisverylowlevel proteins couldbedetectedinthejuice detectable proteins, althoughlowlevelsof red wineswehaveanalysedcontained By HPLC,apartfrom Merlot,noneofthe grapes ofdifferent knownvarietalorigin. white andfivered wineswere vinifiedfrom Pic of Mariola Kwatkowski, Liz Waters & Patrik Jones Patrik Waters& Liz Kwatkowski, Mariola of Pic ough within abatch. wine inbottlescanbeseeminglyrandom this variability, theresulting oxidationof of oxygenpermeationtheyallow. Dueto closure/glass interfacevaryintheamount #666) alsosuggestthatclosures and/orthe Institute’s Closure Trial (Institutepublication publications #534and580).Data from the and/or closure/glass interface(Institute into bottledwinesthrough theclosure for browning isthatofoxygenpermeation additional andmore recently identifiedroute Godden, during theseoperations(Robinsonand problem forwinemakersworldwideand Oxidative degradationofwhitewinesisa Collaborator: Leigh Francis,KateLattey, DrMarkSefton Skouroumounis, MariolaKwiatkowski,Dr Staff: Studies onrandomoxidationofbottledwines differentiate grapes. mass spectrometric methodtovarietally SO operations andbarrel storage.Adequate This canoccurduringnormaltransfer oxidation iscontactofthewinewithoxygen. After fermentation,themaincauseof citrus/lime anddevelopedagoldencolour. age characterssuchashoney, toasty, cooked and citrusnotesbutacquired typical bottle whereas agedRieslingalsolostthe floral became aldehydicandbrown incolour, Riesling lostitsfloralandcitrusnotes been foundinarecent studythatoxidised by oxygen.Asanexampleofthis,ithas being primarilycausedbyreactions promoted result ofvariouschemical reactions, by It differs from ageing,whichislargelythe notes, andthedevelopmentofbrown colour. young wines,namelytheirfloralandfruity results inlossofcharacteristic aromas of 2 levels shouldprevent excessoxidation Dr ElizabethWaters, DrGeorge Mariola Kwiatkowski,PatrikJones&ElizabethWaters Technical Review Andrew Kleinig(SouthcorpWines) 145). One type alsoaffected thefree andtotalSO acid wasnotaddedatbottling.Closure marginally lowerinwinestowhichascorbic at bottling–there wasnoeffect ifascorbic levels dependedonascorbicacidaddition the effect ofstoragepositionontotalSO was lowerinallwinesstored uprightbut acid levelsinwines.Theleveloffree SO was noeffect ofstoragepositiononascorbic were inlinewiththesensorydata.There closures havingthehighest.Theseresults and theROTE(roll-on tamper-evident) having thelowestlevelsofthesecompounds ascorbate levels,withthesyntheticclosure total SO For theChardonnay wines,thefree and Semillon wineintheInstitute’s Closure trial. and Riesling,aresult alsoseenwiththe fresh fruitcharactersinbothChardonnay character were alsonegativelycorrelated with ascorbic acid.Theratingsforoxidised in oxidisedcharacterthanthewineswith ascorbic acidwere ingeneralrated higher more oxidisedaroma. Thewineswithout showed thatthevisuallybrowner wines have were closetothree yearsold.Thisstudy undertaken inAugust2002whenthewines aroma characteristicsofthesewines was A secondlargesensoryinvestigationonthe than thenaturalbarkclosures. synthetic closuremuchlessaffectedbyposition with the colour dependedontheclosuretype, extent oftheeffectstoragepositiononwine The highest ratingsforbrownandorangecolour. the with gavewines general, in ideal conditions, upright storagefortwoandahalfyears The absenceofascorbicacidatbottlingand between winecolourandA420nmvalues. wine, agoodcorrelation wasobserved acid. However, whencomparingthesame comparing wineswithandwithoutascorbic valid asameasure ofbrown colourwhen that A420nmmeasurements were not after twoandahalfyears’storageconfirmed A descriptiveanalysisstudyofwinecolour publication #731). spectr modified throughout theirstorageperiodusinga for eachtreatment have A 420nmonatleast30replicate bottles different suppliers.Measurements of two Reference 2naturalcorksfrom ROTE (roll-on tamperevident)oroneof sealed witheitherasyntheticclosure, a were Bottles on theextentofoxidation. ascorbic acid,bottlepositionandwinetype of toevaluatetheeffect Southcorp Wines from experiment undertakenwithassistance were bottledinAugust1999for an A RieslingandawoodedChardonnay wine Oxygen duringbottlingandstorageofwhitewines influence itsoxygenpermeability. closure andclosure/glass interfacethat contact occurs,andtheparametersof contact withwineanditseffects ifoxygen and storageprocedures tominimiseoxygen The aimsofthisproject are toidentifybottling 2 levels afterthree years’storagewere ophotometer (Institute been taken under 2 and 2 2 The SO weeks afterbottlinginlateJanuary 2003. levels reduced. Afurther checkonSO A checkontheSO Laboratory (WSL). Science Hickinbotham RoseworthyWine The winesare beingstored uprightatthe bottling wastypicalforwinesofthisstyle. respectively. Thecompositionofthewineat insertion andanullageof6mL5.4mL, and asyntheticclosure usingvacuum and sealedwithnaturalcork(Reference 2) a comparison,thesamewinewasbottled with headspacesof4,16and64mLair. As 16 December2002withascrew cap(Auscap) Andrew Kleinig’s assistance,wasbottledon andwithDr provided bySouthcorpWines, A commercial Sauvignonwine Cabernet under thescrew capeffects thesedifferences. and toevaluatewhetherheadspacevolume than undernaturalcorkorsyntheticclosures aroma, altered colourandsensoryproperties under screw capdevelopsmore reduced gather informationaboutwhetherared wine A preliminary experimentwasinitiated to Oxygen duringbottlingandstorageofredwines (Institute AnnualReport2000). was littledifference betweenthewines analysed aftersixmonths’storage,there ascorbic acidwasaddedatbottling.When for twoandahalftothree yearswhen oxidised afterstorageunderidealconditions data allindicatethatthewineswere less conditions, aroma, colourandcompositional under commercial conditions.Under these Chardonnay winebottledonalarge scale ascorbic acidadditiontoaRieslingand has allowedustoexaminetheeffect of study, through collaborationwithawinery, aligned tocommercial practice.Thecurrent the experimentswere notalwaysclosely previous experimentsandtheconditionsof however, hasrarely beenconductedinthese was added.Formalsensoryassessment, and modelsolutionstowhichascorbicacid enhanced depletionofsulfurdioxideinwine publication #595)andothersindicatedan scale workfrom theInstitute(Institute for debateintheliterature. Previous laboratory oxidation ofwinecontinuestobeamatter The effect ofascorbicacidadditionon Research. Journal ofGrapeandWine written upforpublicationinthe Roadshows (seeAppendix1)and will be information waspresented atthe2002 ascorbic acidwasaddedatbottling.This for twoandahalftothree yearswhen oxidised afterstorageunderidealconditions closures examined,thesewineswere less from thistrialclearlyindicatethatforallfour In summary, thechemicalandsensorydata wines. results were observedfortheRiesling acid hadbeenaddedatbottling.Similar increased underthescrew cap,theSO very similar. Astheheadspacevolume the screw capwitha4mLheadspacewere bark closure, thesyntheticclosure and with 2 levels inwinessealedwiththenatural 2 levels occurred six Australian 2 2 SO 4 mLheadspace,intermediateandsimilar the winesealedwithscrew capwitha stage, thehighestSO months afterbottling.Atthisveryearly levels wasundertakenatfiveandahalf Research Australian JournalofGrapeandWine this methodhasbeenpublishedinthe wine isinthebottle.Apaperdescribing the result ofoxygenpermeation,whilstthe has beenmodifiedtomeasure wine colour, As reported previously, aspectrophotometer The inherentpermeabilityofcorkstooxygen wines willbeassessedagainatalaterdate. at thisearlystageofdevelopment.These wine sealedunderthenaturalbarkclosure judged bythepaneltobedifferent from the wine sealedwiththesyntheticclosure were wines sealedunderthescrew capor the with theothertreatments. Noneofthe with naturalbarkclosure were compared these observationsandthewinessealed testing foraroma wasundertakentoconfirm to alltasters.Formalduo-triodifference reduced aromas were notclearlyapparent the treatments maynotbeobviousand again suggestedthatdifferences among bottling inmidJune2003,informalassessment treatments. Fiveandahalfmonthsafter difficult toseedifferences amongthe mL headspaceunderscrew cap,anditwas in anywines,includingthewineswith4 early stage,reduced aromas were notobvious assessed thewinesaftersixweeks.Atthis A tastingpanelattheInstituteinformally screw capwitha64mLheadspace. lowest levelsinthewinesealedwith the screw capwitha16mLheadspaceand with naturalcork,syntheticclosures and from ENSBANA,Dijon,France. of ChristopheGuirado,avisitingstudent the experimentalconditionswithhelp made ondevelopingtheassayandoptimising oxygen. Goodprogress thisyearhasbeen acts asawater-soluble trapofsinglet develop anassayusingacompoundthat our current andlongstandingeffort isto available orsimpletodevelop.Thebasisfor closures inwinebottlesare notreadily low levelsofoxygenpermeatingthrough Chemical assaystoquantifytherelatively 2 levels were seeninthewinessealed (Institute publication#731). 2 levels were seenin 31 Research Teams’ report 32 Research Teams’ report covered closures forthesyntheticclosures. shown bywinesinbottleswiththefully for thenaturalclosures andhigherthanthat the bottleswithfullycovered closures with the‘windows’wassimilartothatin browning ofwinesinbottleswithclosures interface?). Afterninemonths’storage,the (through theclosure ortheclosure/glass route ofoxygenpermeationintowinebottles of thisexperimentwastoinvestigatethe closure forfourreplicate bottles.Theaim ‘window’ remained inthecentre ofthe or topartiallycovertheclosure sothata cover theclosures oftworeplicate bottles, glass andtheclosure toeithercompletely was appliedtothetopsofbottlesover well knownforitsoxygenimpermeability, As previously reported, araldite,amaterial that relate directly tocommercial use. be assessedinarangeofstorageenvironments performance ofclosures inwinebottlescan being inagaseousenvironment. Thus the contact withawine-likesolutionratherthan available systemsisthattheclosure isin of thismethodoverothercommercially within fourtoeightweeks.Theadvantage us toestimateoxygenpermeationrates indicates thatthisassaymayindeedallow up inFebruary2003.Datacollectedtodate bottles sealedwithasyntheticcorkwasset to monitoroxygenpermeabilityintowine A preliminary experimentusingthe method covered. ItislikelythatSO colour toberelated tothearea oftheclosure however, asexpected,thetrend isforwine all thetreatments havebecomeevident, differences inA420measurements among So far, afterninemonths’storage, noclear are nowbeingmadeonceeverysixweeks. then fortnightlyuntilthefifthmonthand were madeeveryweekinthefirstmonth colourmeasurements (A420nm, Wine Institute. Thewinesare stored uprightat25˚C. araldite treatmentsand appliedatthe then the with thehelpofAndrew Kleinig Southcorp at bottles.Thewineswere bottled replicate (no aralditecoverage)hasatleasts araldite treatments includingthecontrol nine the of for thisexperimentandeach used been has cork orasyntheticclosure) sealed withoneoftwoclosures (a natural wine One 50% oftheclosure asasemicircle. or asaringaround theedges,orto cover closure, either asacircle inthecentre ofthe 50% or20%ofthesurfaceclosure been usedtoeitherfullycover, cover80%, on theseearlierobservations.Aralditehas has beensubsequentlysetuptobuildfurther A more extensiveexperimentusing araldite differences becomelarge. be depletedsufficiently before colour 2 levels needto even in situ Gayle Baldock&Yoji Hayasaka ) service provider tograpeandwineresearch, collaborate withProvisor, anew technical problem solvingissues.Ourfuture abilityto contributions tootherprojects andarising current projects andtoprepare forfuture necessary toenhancetheprogress of the The additionalelectrospray capabilityis electrospray capability. the MSfacilityneedstohaveadditional the increase indemandforthisapplication, tool forproblem solving.Inresponse to and tanninprojects andhasbecomeauseful indispensable toolfortheInstitute’s protein reporting period.Thistechniqueisan technique hascontinuedtoincrease inthe The applicationoftheelectrospray Mass spectrometryfacilityusagetrends including publicinstitutions. and expertisetothescientificcommunity advanced massspectrometric techniques (iv) asaprovider oftheversatileand activities involvingmassspectrometry and Adelaide intheresearch andteaching (iii) asacollaboratorwithTheUniversityof makers, usingmassspectrometric techniques; facing thewineindustryandindividual (ii) asaninvestigatortosolvetheproblems spectrometry tograpeandwineresearch; act; (i)asaleaderintheapplicationofmass Campus MassSpectrometry Facilityare to The fourimportantroles oftheWaite Staff: Waite CampusMassSpectrometryFacility Yoji HayasakaandGayleBaldock will give us access to new equipment Table 6. ‘Problem’ investigations conducted in the period from July 2002 to June 2003 including electrospray mass spectrometers. It represents an opportunity for us to Type of investigation Number of samples analysed overcome the current and growing problem of limited mass spectrometric resources. Brine contamination (Alcohol based) 35 The Facility’s TSQ GC-MS/MS and API LC- Protein analysis for varietal identification 30 MS/MS are used for various purposes and Brine contamination (Glycol based) 9 appropriate financial arrangements for all users are in place to recover the running Unknown taint 8 cost of the Facility. The usage of the TSQ Closure problem 6 GC-MS/MS and API LC-MS/MS for the year Diesel analysis 4 is detailed in Table 5. Paint/varnish analysis 3 Collaboration with the Industry and Hydraulic oil analysis 3 Analytical Services Kerosene analysis 1 During the reporting period, the Facility Sorbic acid analysis 1 conducted mass spectrometric analyses on 100 non-research samples, in collaboration with staff of the Industry and Analytical Total 100 Services. The analyses conducted during the reporting period are detailed in Table 6. Brine contamination quantification limit of propylene glycol Juice protein analysis for a varietal identification using this method is 10 mg/L. Therefore Of all ‘problem investigation’ analyses the method is capable of detecting It is highlighted that this new technology conducted during the year, the juice and wine approximately 0.001% glycol in wine (10 delivered from our research has been adopted samples suspected to be contaminated with mL of glycol in a tonne of wine). for use by Industry Services in consultation brine accounted for 44% of the total analyses with winemakers who submitted 30 samples conducted by the Facility. In response to an Collaboration with research groups to the Institute for protein analysis during increasing demand on the analysis of brine, this financial year. This new technology is the Facility developed two reliable methods The contribution of the MS facility to the Institute’s research projects is detailed in the reports under Wine grape tannin and colour Table 5. Usage of the TSQ GC-MS/MS and API LC-MS/MS for 1 July 2002 to 30 June 2003 specification and Studies on unstable wine proteins involved in haze formation. TSQ GC-MS/MS API LC-MS/MS Briefly, the following outcomes are highlighted: The Australian Wine Research Institute 91.0% 77.3% In collaboration with Dr James Kennedy The University of Adelaide 9.0% 20.7% (Oregon State University, USA), the direct CSIRO 0.0% 1.4% condensation product (polymer) of an anthocyanin and proanthocyanidin was Flinders University 0.0% 0.6% characterised using chromatographic and mass spectrometric techniques. The degree of polymerization was found to be as high used for varietal differentiation of juice for the detection of alcohol-based and as an octamer composed of an anthocyanin based on the composition of pathogenesis glycol-based brine, respectively. combined with a proanthocyanidin heptamer. related-proteins (PR-proteins) (Institute This is the first mass spectrometric evidence publications #663 and 734). This study (1) Alcohol-based brine confirming the existence of the direct described that PR-proteins in untreated The red pigment, rhodamine is one of the condensation products of an anthocyanin juice were detected using electrospray mass components in alcohol-based brine. It is with proanthocyanidins since Somers spectrometry combined with a protein trap present at a concentration of 10 mg/L but this proposed the formation of ‘pigmented cartridge (Trap-MS method) and the resulting concentration may vary among manufacturers. polymer’ in aged wine in 1971. protein composition in the juice was Contamination with alcohol-based brine was consistent among samples representing the evaluated by the ‘presence’ or ‘absence’ of In collaboration with Dr Stéphane Vidal and same variety but differed among varieties. rhodamine in juice or wine and then the Dr George Skouroumounis, the direct The method was validated using juices from degree of contamination was estimated by condensation products of anthocyanins were berries from 20 different varieties (Vitis its concentration. Rhodamine was detected characterised using MLCCC and mass vinifera cv.) harvested in at least two different using electrospray LC-MS in selected ion spectrometric techniques. The presence of the seasons and from different vineyards. reaction mode after the extraction and direct condensation products of anthocyanins enrichment of rhodamine using a C18 in either grape skins or wine has not been The analytical results of the submitted cartridge. This method is capable of detecting reported. This is the first mass spectrometric samples demonstrated that the technology 0.001% brine in juice or wine, which is evidence to propose the existence of could be used as an additional tool for equivalent to 10 mL of brine in a tonne of anthocyanin oligomers in grape skins and wine. winemakers to ensure the varietal origin of juice or wine. juice. The availability of this technology is The switchability of the mass spectrometric expected to place Australian winemakers in (2) Glycol-based brine technique for varietal differentiation based a stronger position than their overseas A GC-MS method was developed mainly on PR-proteins of both white and red grape competitors to back up claims of integrity for the detection and quantification of 1,2- juices has been further confirmed. The extent from vineyard to bottle. propylene glycol in red and white wine. to which the discriminating power can be Propylene glycol was reliably quantifiable in extended to white wine is limited if

a range of 10 to 250 mg/L. The bentonite fining has occurred. report Teams’ Research 33 34 Research Teams’ report > Committee includes: The activityoftheProvisor Building Building Committee. Provisor asmembersoftheProvisor participated intheestablishmentof Yoji HayasakaandDrAlanPollnitzhave Contribution totheestablishmentofProvisor > > > > To investigatethemeritsof computer To discussmattersinrelation toanew To specifyandassesstheexisting To recommend thespaceandservice To identifyandprocure theinstruments building forProvisor. can berelocated toanewbuilding. instruments oftheshareholders which for labsandoffices. (gas, water, electricityetc)requirements service activities. which are required fortheProvisor AWRI andCSIRO. Provisor, between network integration and TA. although there were somedifferences in pH such asalcoholwasfoundtobevery similar composition ofeachsampleformeasurements had somevariationinTSSatharvest. The comparable, asthesetprevious year the gross compositionofthewineswas addition, great care wastakentoensure that harvested fruitinacommercial situation.In imposed tomimictransportofmachine but, inthiscase,aperiodofskincontactwas ashadbeendonepreviously mildew infection different four levelsofpowdery into sorted wine wasmadefrom fruitwhichhadbeen is partofCRCVProject 1.5.2.Chardonnay with winefrom the2002vintage.Thestudy The UniversityofAdelaidewascarriedout with BelindaStummerandEileenScottof Institute Publication#714),asensorystudy Research and Wine published inthe Following from previous work,recently Eileen Scott,TheUniversityofAdelaide Collaborators: Staff: Powdery mildewstudy Dr LeighFrancisandKateLattey Kate Lattey &LeighFrancisKate Lattey Dr BelindaStummerand Australian JournalofGrape (2003, 9,28-39, lowered fruitattributes. these attributes,withconcomitantly grapes beingratedthemostpronounced in made from themostseverely affected compared tothecontrol, withthewine rated ashigherinfungalandearthyaroma addition, the‘infected’treatments were was asfoundintheprevious study. In measures inthewines.Thisobservation attribute wascorrelated withphenolic control, uninfectedtreatment, andthis viscosity/oily attributecompared to the perceived tobesignificantlyhigher in a even atthelowestlevelofinfection,were made from powderymildew-affected fruit, The sensorydataconfirmedthatthewines Industry Services Teams’ report Technical problem solving and consulting as educational, seeking to disseminate years is presented in Table 7. The number information in a variety of ways in order to of investigations conducted decreased by Staff: Peter Godden, Adrian Coulter, Mark foster industry wide understanding of the 8% from the previous year. However, the Gishen, Ella Robinson, Peter Høj, Geoff causes of many common winemaking total number of samples analysed increased Cowey, Yoji Hayasaka, Trudy Wallis, Greg problems, in order to prevent their frequent by 89%. Twenty per cent of this increase Ruediger and Peter Valente recurrence. The Industry Services team also is accounted for by samples analysed as provides technical support to the Institute's part of the investigations into the effects of The Industry Services team provides technical Analytical Service, particularly in the bushfire smoke on grapes and wines, which development and support services to the maintenance and auditing of the quality are described below. Notwithstanding the Australian wine industry, primarily in the form management system, and the interpretation bushfire smoke investigations, the of an advisory service that disseminates a of analytical results. The Analytical Service previously reported trend of a greater wide range of technical information, and a also supplies chemical analysis on problem number of samples being analysed for each problem solving and analysis service, which solving and research samples to the Industry investigation has continued. This trend has collectively represent a significant proportion Services team, on a contractual basis. been driven by a number of factors, including of the team's workload. The team continues an apparent increase in the use of Industry to manage a large research trial that is The Institute's investigative and advisory Services reports to support insurance claims, examining the technical performance of services are provided according to and in legal proceedings. More recently, it various types of wine closure, and a project strict Terms and Conditions, and client has become a National Association of that has developed a web-based technical confidentiality is an important aspect of the Testing Authorities (NATA) requirement of reference manual for the Australian wine provision of the services. This facilitates a the Analytical Service laboratory that industry, with associated workshops that frank exchange of information between measurements of the uncertainty of the are presented in conjunction with Institute the Institute and its clients, which in turn results of analyses be reported with each Roadshow seminars. In addition, the team allows the maximisation of the knowledge result. It is considered good practice for this coordinates an Institute-wide project that is gained from the provision of these services. policy to also be adopted by the Industry investigating the relationships between the Services laboratory, and in its reporting, in potential spoilage yeast Dekkera/Brettanomyces A proportion of the investigations conducted order to strengthen the statistical certainty and wine in Australia. by the team relate to disputes arising between of conclusions that may be drawn in these levy-payers or between levy-payers and reports. Consequently, due to the inherent The Industry Services Laboratory analysed suppliers of either materials or contract degree of uncertainty of many of the in excess of 2000 samples during the year, services. Consequently, and somewhat analytical methods used, it is clear that a using a wide range of routine or novel reluctantly, Industry Services staff members, large number of samples need to be assessed analytical techniques. An increased reliance with the involvement of the Director and in order to obtain the desired level of on advanced analytical methods provided Company Secretary, often find themselves statistical significance. by the Waite Campus Mass Spectrometry in a mediation role in these disputes, and Facility, the Institute’s commercial Analytical spend a considerable amount of time Approximately 40% of investigations Service and our sensory panels led by Kate providing technical information to legal conducted during the current year related Lattey and Leigh Francis is acknowledged. professionals representing grapegrowers to the combined problems of hazes and The majority of samples analysed are wine; and wine companies. It is, therefore, clear deposits and microbiological instabilities. the analysis of which is supplemented by that the support offered to the Australian Whilst the number of wines exhibiting these detailed sensory evaluation by a panel of wine industry by the provision of these types of problems as a proportion of all experienced tasters. The remaining services is increasing in scope, and is of wines made in Australia appears to have samples predominantly consist of wine great benefit to clients, as the majority of fallen in recent years, the overall number additives, closures, or compounds that are disputes are settled before formal court of wines affected should remain a concern suspected to have caused taints and or proceedings are instigated. for the Australian wine industry as a whole. deposits in wine. The capability of the Issues related to such instability problems Industry Services team was enhanced during A summary of the number and type of continue to be addressed by GWRDC- the year by the recruitment of Geoff Cowey investigations conducted by the Industry funded Project 99-1 Targeted training of (formerly employed in a large wine company Services team over the past three financial wine industry personnel: Compilation of a as Chemist/Microbiologist) in the position of Chemist – Industry Services, and by the Table 7. Summary of the number and type of problem solving investigations conducted, and numbers of samples analysed team gaining access to a new GWRDC by Industry Services during the past three years funded GC-MS instrument. Investigations conducted and samples analysed The primary aim of the investigative and 2000/01 2001/02 2002/03 advisory service is to offer preventative and remedial advice based on the cumulative problem solving experience of the staff, and Identification of hazes and deposits 135 85 112 the practical winemaking experience of the Microbiological investigations 50 89 95 team Manager, Oenologist and other staff members, rather than providing simple Sensory assessments 51 98 89 diagnoses of the causes of various problems. Taint problems 66 43 72 It is clear that quality loss during wine processing and packaging represents a Other investigative analyses 128 209 113 major cost to the Australian wine industry, Closure-related investigations 24 22 20 and consequently all the activities of the Industry Services team, in terms of problem solving, extension and information transfer, Total number of investigations 454 546 501 and research, can increasingly be seen to address this issue in a targeted manner, such as exemplified by the closure trial and Total number of samples analysed 1048 1233 2231 more recently our ‘Brett’ project. To this end, staff members increasingly regard their role Industry Services Teams’ report Teams’ Services Industry 35 36 Industry Services Teams’ report > > effectiveness ofSO practical informationonoptimisingthe Services staff produced anarticleproviding has increased. Consequently, Industry sulfur dioxide(SO insufficient concentrationofthepreservative in whichtheapparent primarycauseisan the proportion oftheproblems investigated in more detaillaterinthisreport. Additionally, of associatedworkshops technical referencemanualanddevelopment investigated thisyear. interesting andunusualcaseshavebeen extremely varied,andsomeparticularly 7 as'otherinvestigativeanalyses'are The typesofinvestigationsrecorded inTable in thisarea. by theInstitute'severincreasing capabilities for thistypeofworkcanbepartlyexplained previous year, andthecontinueddemand close totherecord highlevelrecorded inthe conducted during2002/2003hasremained The numberofsensoryassessments issue 145(August2003). wine, forpublicationin A brown granulardepositisolatedfrom a Following apublicationinrefereed by thefactthatitwasfoundtobe soluble metal inthedepositwasalsosuggested the presence ofcalcium. produced anorange-red flame, s however, aflametestonthematerial in theinfra-red spectrumoftheisolate, characteristic absorbanceswere observed amorphous material.Nosignificantor revealed lightbrown crystalswithsome Microscopic bottle ofbrandywasinvestigated. purported tobe. were 100%ofthevarietythattheywere the protein analyseswasthatthejuices In bothcases,theconclusiondrawnfrom from theleesthathadbeendespatched. juice-lees filtrationwasinfactthefiltrate returned toawineryfollowingcontract been had another, whetherjuicethat fact ‘lowripenessSem Blanc juicethatithadpurchased was in whethersomeSauvignon questioning assessment ofjuice,awinerywas sensory publication #663).Inonecase,basedon Hayasaka et.al.2001(Institute Peter Høj,according tothemethodof Yoji Hayasaka,andtheDirector, Professor Institute’s MassSpectrometry Manager, the of samplesthatwere analysedby process. TheInstitutereceived a number winemaking has occurred duringthe other,fearswinemaker a a rare mix-up particularly usefulif,foronereason orthe measures intheindustry. Thiscanbe measures tostrengthen labelintegrity winemakers keentoimplementfurther has beenengagedbyanumberof descriptors forgrapejuice,theInstitute the feasibilityofauthenticatingvarietal synopsis inour (Institutepublication#663)anda journal examination ofthematerial Technical Review 2 ), particularlyatbottling, 2 additions madeto illon’, andin Technical Review , whichisdiscussed The presence of uggesting regarding > As reported inprevious AnnualReports, GC-MS andLC-MSmethodsbythe other The developmentofthisandseveral commercially availablerefrigerant brines. rhodamine, whichisusedasadyeinseveral GC-MSanalysisforthecompound of a been greatly aidedbythedevelopment has and theinvestigationofthesesamples having occurred inrelatively largewineries, current year, duetobrineleaksapparently refrigerant brinecontaminationduring the samples wasinvestigatedforpossible heeded. Ahigherthanusualnumberof presented intheseworkshopshadbeen could havebeenavoidediftheadvice of thecasesinvestigated,contamination Roadshows, anditisapparent thatinmany the workshopspresented duringInstitute contamination continuetobeaddressed in minimisation oftherisksthistype care toconsumers.Issuesrelated tothe an integralstepintheindustry’s dutyof occurred accidentally. Suchprecaution is unintended introduction ofsubstanceshas one reason orother, itissuspectedan wine totheInstituteasaprecaution if,for type ofproblem isduetowineriesreferring aware thattheInstitute’s contactwiththis brine andhydraulicoil.Readersshouldbe contamination continuetoberefrigerant andthemostcommoncausesof concern, not permittedadditivestowineisof contaminated withsubstancesthatare wines thatare foundtobeaccidentally washed filterpads(Institutepublication#58). insufficiently for dilution,andleachingfrom are water used sources ofcalciuminbrandy #58). The main publication (Institute mg/L concentration ofcalciumisgreater than5 t as theamorphous beenknowntoprecipitate brandy and has elements causinginstabilityin important years. Calciumisoneofthemost 14 least at instability investigatedattheInstitutefor the firstdocumentedcaseofthistype of concludedtobecalcium tannate, therefore, condensed t red colourindicativeofthepresence of for 30minutes,whichproduced abright and hydrochloric acidandheatedat80°C was dissolvedinamixture ofmethanol 28% calcium.Aportionofthematerial spectrometry indicatedthatitcontained material usingatomicabsorption in 10%hydrochloric acid.Analysisofthe annins. Theisolatedmaterialwas, annate whenthe Geoff Cowey, AdrianCoulter&EllaRobinson > > An investigationwasconductedintoared One winesampleanalysedwasfoundto ftewn suggestedthatthecombination of the wine oftheresults ofchemicalanalysis Inspection bacteria, possibly possibly have beeneitherlacticacidbacteria, may sp., aswellrod-shaped bacteria which Saccharomyces viable yeastcells,suspectedtobeof deposit thatwasfoundtoconsistofnon- occurred. Thewinealsocontained a week period,aswasreported tohave concentration ofVA inawineovertwo increase ofapproximately 0.85g/Linthe for however, itwouldbeconsidered unusual of LABiscapablecausingspoilageinwine, suspected tobe it containedviablelacticacidbacteria(LAB), Microbiological analysisofthewine indicated increase inVA inthiswine. discounted asacauseofthedramatic oxidation ofethanoltoaceticacidwas chemical the concentration ofacetaldehyde, excessive brown colouror an el the winesuchasdevelopmentof of othermarkerschemicaloxidation risen to1.39g/L.Intheapparent absence weeks latertheconcentrationofVA had was 0.52g/L.However, approximately three period theconcentrationofVA inthewine further eightweeks.Attheendofthis then subjectedtomicro-oxygenation fora sulfur dioxidewasaddedandthewine (MLF). AfterthewinehadcompletedMLF, while undergoingmalolacticfermentation subjected tomicro-oxygenation for14days been had in ashorttimeperiod.Thewine (VA) hadreportedly increased dramatically acids wine inwhichtheconcentrationofvolatile closure trialwasestablishedtobe1ng/L. Semillon wineaspartoftheInstitute’s the experiencedtasterswhoassessedawhite recognition threshold forthiscompound AnalyticalService.Thesensory Institute’s concentration everrecorded bythe contain 1860ng/LofTCA,thehighest Their workisappreciatively acknowledged. Services teamduringthereporting period. Baldock, hasgreatly aidedtheIndustry Yoji Hayasaka,andtechnicalofficer, Gayle Institute’s MassSpectrometry FacilityManager, Lactobacillus Lactobacillus Lactobacillus sp. and sp. bacteriatocausean Acetobacter sp., oraceticacid Brettanomyces sp. Thisspecies sp. evated of moderate SO2 concentration (the variable microorganisms in the form of a The Industry Services team considers that

winemaker had made an SO2 addition deposit. Following microbiological analysis this issue was the single largest problem when the increase in VA was discovered, of this first sample, an informal sensory dealt with since the inception of the team, and prior to sample being submitted to assessment of the wine was conducted and both in terms of its value and the numbers the Institute), and low pH, may have been it was considered obvious that the wine of wine companies and grapegrowers responsible for the lack of viability of the contained a high concentration of volatile affected. The investigations began with microorganisms found in the deposit. acids (high VA). This sample contained a ‘tide approaches from technical staff of several line’ in the form of a ring of dark coloured companies in Victoria and South Australia, Brettanomyces yeast are capable of causing material adhering to the inside surface of the who considered that there was a problem spoilage in wine and may have been neck of the bottle, which was found to with ‘smoke taint’ in fruit that they had responsible for the observed increase in the consist of rod-shaped bacteria, suspected either processed or had contracted to concentration of VA if the yeast had been to be Acetobacter sp., coccoid bacteria, purchase. Over subsequent days the number viable at the time that the increase occurred. suspected to be Oenococcus sp., and yeast of approaches to the Institute became If viable Brettanomyces yeast were present cells, suspected to be Saccharomyces sp. The overwhelming, and it was clear that a in the wine before MLF, then it is possible wine was found to contain a concentration of dedicated trial was required to properly that the population of the yeast may have volatile acidity of 1.9 g/L, and the results understand the nature and extent of the increased during MLF, aided by the presence of microbiological analysis of the wine problem, rather than a ‘scatter gun of oxygen due to micro-oxygenation. indicated that it contained viable rod-shaped approach’ of working with several Subsequently, following further micro- bacteria, suspected to be Acetobacter sp., companies concurrently. The Institute, oxygenation, the population of the yeast as well as viable lactic acid bacteria and therefore, supported a move by the Alpine may have reached a level where they were yeast (see also Institute publication #718). Valley Winemakers and Grapegrowers able to produce the observed increase in Association to fund a researcher to conduct the concentration of VA in a short period Observations of the linings of the various trials. The Industry Services team shaped bacteria observed in the deposit screwcaps showed perforated rings helped to design targeted trials, much of were acetic acid bacteria, and that these around the area of contact with the finish of this work necessarily being performed over bacteria were responsible for the increase in the bottles, with some ‘puckering’ of the a very short time frame. It is considered the concentration of VA, if it is assumed liner material. Although the linings inside that the approach taken by the association’s that they were viable following MLF. As with the caps of all of the bottles appeared to be nominees and other affected parties was the former scenario, micro-oxygenation of damaged to some extent, ‘tide lines’ existed extremely thorough and professional, such the wine may have facilitated an increase in in only some of the bottles. These bottles that well controlled and executed replicated the population of acetic acid bacteria if they were found to exhibit higher levels of experiments were conducted, and well

were present and viable during the process. volatile acidity, little or no free SO2, and prepared samples delivered to the Institute exhibited high turbidities compared to for analysis. The fact that the affected It is difficult to determine which samples with no ‘tide lines’. The wine in regions involved declared phylloxera zones microorganism or organisms were six bottles that did not exhibit ‘tide lines’ complicated the preparation and freighting responsible for the increase in VA in this contained concentrations of volatile acidity of samples to Adelaide, and at all times wine, although it is possible that the process of between 0.53 and 0.56 g/L. The wine in phylloxera control protocols were followed. of micro-oxygenation may have contributed five other bottles that did exhibit ‘tide lnes’ The Institute also acknowledges the valuable to the problem. It is the standard advice of ranged from 0.86 to 1.21 g/L. These input and assistance provided by John Industry Services staff that wines undergoing observations suggest that some of the Whiting of the Victorian Department of micro-oxygenation should be sensorially closures had apparently been damaged to Primary Industries, Jill Kuchel of Vignoble assessed regularly, preferably on a daily basis. such an extent that they had allowed air to Monitoring Services, Wendy Cameron and The results of this investigation suggest that enter those bottles. This had resulted in Terry Barnett of Brown Brothers Milawa

this advice should be extended to include the the loss of SO2, thus allowing the growth of Vineyards, and Shayne Cunningham of weeks immediately following the cessation of spoilage microorganisms that had produced Gapsted Wines. As a result of various micro-oxygenation. We are not concluding the elevated concentrations of acetic acid, trials, it is considered that a high level of that the application of microoxygenation and caused the increase in turbidity and understanding of the problem has been in a correct fashion leads to dramatic creation of the ‘tide lines’ in those bottles. achieved, but unfortunately in spite of increases in VA, however, we believe it which, the problem has proved to be quite important that practitioners are informed A workshop to be staged at the 12th intractable in most cases. of this incident. Australian Wine Industry Technical Conference, which will be held in Melbourne in July The main conclusions of the initial > During the year the Institute became 2004, will examine factors relating to the investigations were: aware of a number of problems resulting correct application, and quality control of from the mis-use of roll-on tamper-evident screwcap closures. > Various wines and juices submitted to the (ROTE) screwcap closures. The problems Institute were indeed considered to ranged from the use of closure and bottle Investigations conducted into the nature and exhibit characters variously described by the combinations that were incompatible, a fact amelioration of taints in grapes and wine, Institute’s sensory panel as smoky, burnt, that was readily identifiable by inspecting the caused by smoke resulting from bushfires ash, ashtray, salami, smoked salmon etc. specifications of the closures and bottles, to three instances where the force used to During the 2003 vintage, Industry Services > It was established that guaiacol and 4- apply the closures appeared to have been staff spent a large amount of time dealing methylguaiacol were the most important too high. In one case, this resulted in the with the issue of ‘smoky’ taint in grapes and compounds contributing to the sensory tamper-evident perforations on the closures wine resulting from the bushfires that occurred taint. Guaiacol and 4-methylguaiacol are being broken on some bottles, or damage in Victoria and southern New South Wales in compounds that commonly occur in to the internal lining of the closures in the January and February 2003. The involvement wines that have been matured in contact two other cases. In one of these cases a of the Institute’s Analytical Service in with toasted oak products, and are formed dozen bottles of a Pinot Noir wine were expediting the analysis of samples as part of during the toasting process from the submitted for investigation of an apparent these investigations is acknowledged, as is degradation of lignin. Importantly, a back- sporadic oxidation problem. One of the the input of other Institute staff particularly palate excessively drying character and a bottles was opened, and was initially Dr Leigh Francis, Kate Lattey, Dr Mark Sefton, lingering retro-nasal ash character appeared subjected to microbiological analysis Dr Alan Pollnitz, Dr Paul Henschke and to be more pronounced in smoke-affected because the sample appeared to contain(BELGIUM)Creina Stockley. samples than in juices or wines spiked report Teams’ Services Industry 37 with similar concentrations of guaiacol. A trial was conducted to ascertain if it was the ability of various fining agents to 'scalp' It is therefore considered likely that other possible to reduce the guaiacol concentration or remove various flavour compounds from compounds resulting from the smoke were in grapes and wine by applying various wine, and a preliminary summary of the present in the juices and wines at albeit very ‘vineyard-washing’ treatments. The treatments results by Institute Research Chemist, Dr Alan low concentrations. However, it was not examined were cold water, cold water plus Pollnitz, was published in Technical Review possible with the resources available to identify wetting agent, warm water (approximately issue 142 (February 2003), and is also discussed the presence of such compounds, their possible 25°C when contacting the fruit), cold water elsewhere in this report. Guaiacol was one concentrations, or their possible sensory or plus 5% ethanol, and milk. None of these of the flavour compounds examined. Of the other impacts. As such, guaiacol and 4- treatments reduced the guaiacol fining agents examined, only activated methylguaiacol cannot be considered as solely concentration in either free run juice or carbon was found to remove any guaiacol, responsible for the identified taint. crushed grapes that were macerated with the concentration being reduced by skins for either one hour or 24 hours. approximately 5% with the addition of 300 > Samples of a number of reference juices Guaiacol concentrations were found to mg/L of carbon. The effect of colloidal silica obtained from The University of Adelaide’s increase with increasing maceration time, was not examined in the Pollnitz study, and Hickinbotham Roseworthy Wine Science and samples macerated with leaves consequently the action of this fining agent Laboratory (WSL), which had been sourced contained higher guaiacol concentrations. This was tested on a smoke affected Sauvignon from various vineyards around Adelaide finding supported empirical observations made Blanc juice that had been found to contain where there had not been bushfires during by winemakers, that machine harvested fruit was 18 µg/L of guaiacol. Additions of 0.5 and the growing season, were found to contain more badly affected than similar fruit that had 1 g/L colloidal silica were added using Bakesol no detectable guaiacol or 4 methylguaiacol. been hand picked, and further empirical 30, and the samples were shaken for two The assistance of Mr Stephen Clarke in observations that the free-run juice from whole minutes, and then allowed to stand for one providing these samples is acknowledged. bunch pressing was less badly affected than the hour before being filtered and analysed for Mr Phil Spillman (pers. comm.) informed pressings, or juice from similar fruit that had guaiacol concentration. The concentration the Team Manager that in his trials been machine harvested. Samples of the of guaiacol was not affected by either fining previously conducted at the Institute using liquids from each of the washing rate. The intensity of smoky or ashy taint was a Cabernet Sauvignon wine sourced from treatments were also collected and not sensorially assessed in these samples. the 1993 vintage in Coonawarra, a control analysed. While it was obvious that some wine that was stored in stainless steel for of them contained particulate matter, little It was considered that an explanation was 93 weeks contained 5 µg/L of guaiacol, or no guaiacol was detected in them. The required as to why the vineyard washing which was attributed to the hydrolysis of Ovens Research Station (Department of treatments had no effect on guaiacol and fruit-derived precursors over time. Primary Industries [DPI] Victoria) examined 4-methylguaiacol, compounds that are bunches and leaves from the trial and reported to be very soluble in water. It was > The concentrations of guaiacol and 4- ascertained that approximately 90% of ash considered important to understand the methylguaiacol in various sets of juice and and particulate matter had been washed location of the guaiacol within or on the wine samples were strongly correlated with off the grapes by the water washing grape, in order to assist the minimisation the overall sensory panel rating of the treatments. As it is possible that this ash of its extraction during processing. intensity of the taint. It is important to note, contained compounds with an undesirable Therefore, various samples of grapes were however, that this does not imply that guaiacol sensory or oenological impact, it was peeled, and the skins and pulp were and 4-methylguaiacol are solely responsible for deemed preferable to remove it before analysed separately. The results of this the taint, and no work was performed to harvesting by applying a water spray in the experiment were as follows: investigate any other compound that might be vineyard. present in affected juices or wines. > Guaiacol and 4-methylguaiacol were These results from this initial vineyard detected in all of the skin samples, but > Little information is available in the literature washing trial were available within four days were not detected in any of the pulp concerning the sensory thresholds of of the instigation of the trial, and based on samples. guaiacol in juices and wines. The sensory them the following advice was developed: difference threshold for guaiacol in white > In subsequent experiments, washing bunches juice was established to be 6 µg/L or less, > Leaf plucking, followed by a high-volume, in 95% ethanol for 30 seconds had no using a sensory panel comprised of tasters high-pressure cold water wash in the vineyard, effect on the concentration of guaiacol in who had previously been exposed to albeit followed by hand picking and whole bunch crushed grape samples that were macerated comparatively small numbers of affected pressing with the separation of juice into with skins for 24 hours. The ethanol that samples. It is possible that tasters with more several press fractions, was most likely to had been used for washing subsequently experience in the identification of the taint minimise the taint and allow maximum contained a very low concentration of would have demonstrated a lower threshold. value to be salvaged. guaiacol equivalent to approximately 4 µg per bunch. Similar results were obtained Using the same tasting panel, the sensory > In the event that hand picking was not an from an experiment using hexane as the difference threshold in a red wine that option, then leaf plucking followed by a solvent, rather than ethanol. contained a background level of 37 µg/L of high-volume, high-pressure cold water guaiacol was established to be between wash in the vineyard, and minimisation of One purpose of this experiment was to approximately 15 and 25 µg/L. It is possible leaf matter entering the fruit during ascertain if the guaiacol was located within that in a wine with a lower background harvesting should be employed. the wax bloom on the grapes, or whether concentration of guaiacol the sensory it was partitioned in the skin. Based on the threshold would also be lower. Some of the This advice was actively disseminated to assumption that the ethanol and hexane did bushfire smoke affected red wines analysed growers and wine companies via various remove the wax bloom from the grapes, contained in excess of 70 µg/L of guaiacol. grower and winemaker associations, DPI the results suggest that the guaiacol had The Institute’s Analytical Service database Victoria, and many individual contacts made permeated the grape skin, but that it had contains results of several hundred guaiacol with the Institute by growers and media not passed through the skins into the grape analyses. This sample-set must be considered outlets (see Table 13). pulp. It is suggested that this experiment as potentially heavily skewed towards should be repeated using chloroform, as wooded wines and wines that formed part Investigations were also conducted in order previous workers have used this solvent in of various commercial cooperage trials. to ascertain if it was possible to reduce or experiments to elucidate the composition Approximately 60% of these samples ameliorate the taint during winemaking. A of the wax cuticle on grape skins (Martin contain guaiacol concentrations below 20 µg/L. current Institute research project is examining 1960, and Radler 1965). 38 report Teams’ Services Industry grape growing areas, there is a possibility Winemaking consultation Based on the knowledge that the guaiacol that smoke taint might become a sporadic appeared to be located in the grape skin, it but more common occurrence in the future. The Industry Services team provides a was considered reasonable to predict that It is evident that few vineyards were actually winemaking consultancy service principally the maximum extraction of the compound damaged by the 2003 fires, but damage through the Manager, Peter Godden, a would occur during red wine maceration in caused by smoke taint was widespread. It qualified and experienced winemaker; Adrian the presence of ethanol. is, therefore, apparent that in this situation Coulter, a Graduate in the Diploma in insurance cover for smoke damage would Oenology from The University of Adelaide; A number of batches of red grapes were have been of greater benefit than for fire and Mark Gishen, a Chemical Engineering analysed for guaiacol concentration pre- damage. However, while it appears that graduate from The University of Melbourne crushing, using the 24-hour maceration few growers had insurance cover for smoke with considerable experience in the Hunter treatment that had been used in earlier damage, many do maintain insurance Valley industry. Greg Ruediger, the Trace experiments. Additionally, samples were against contamination. Analysis Laboratory Supervisor, works with homogenized in a 10% ethanol solution in the Industry Services team on a 0.25 basis, order to ascertain if this treatment could be In spite of the fact that this project has and also holds a Graduate Diploma in used to predict, pre-processing, the total greatly increased the understanding of the Oenology from The University of Adelaide. amount of guaiacol that would subsequently nature of this problem, solutions remain Both Adrian Coulter and Greg Ruediger have be extracted during fermentation. elusive. Late in the reporting period, the gained vintage experience with local wineries Fermentations were sampled daily and at Institute analysed a number of commercial during recent years. Further technical advice pressing, where free run and light and heavy samples that were purported to originate is provided by Ella Robinson, who has a BA press fractions were sampled. Samples of from a reverse osmosis treatment of red wines, and BSc from The University of Adelaide, and marc ex-pressing were also analysed. The which was being applied with a view to Geoff Cowey, who has a BSc from The following results were obtained from these reducing the intensity of the taint, and the University of Adelaide. various red wine fermentation experiments: concentration of guaiacol in the wines. The results demonstrated an apparent reduction Most queries received are technical in nature > The concentration of guaiacol increased in in guaiacol concentration of approximately and arise predominantly from Australian a near linear manner for the first three to one third, in the two wines tested. Sensory winemakers. However, many queries are four days of fermentation, and increased analysis was conduced on pre- and post- also received from wine industry suppliers and only slightly thereafter. treatment samples of one of the wines, with Government bodies, as well as a relatively the Institute’s sensory panel rating samples small number from the general public and > Free run, light and heavy press fractions that contained lower guaiacol concentrations secondary and tertiary students. The number all contained the same concentration of as lower in smoke taint. It is possible that of calls received from journalists has increased guaiacol and 4-methylguaiacol. the process may also remove compounds substantially over a number of years (see other than guaiacol that may contribute to Table 13 for list of media enquiries handled > Approximately 25-33% of the total guaiacol the perception of the taint. However, there during the year), and this trend is considered present in grapes was apparently extracted were no significant differences between the to reflect both increasing public awareness when grapes were crushed and macerated panel’s ratings of preference for the samples and interest in the wine industry, and also with skins for approximately 24 hours pre- or post-treatment. While the results of the nature of the projects being pursued by before guaiacol analysis was conducted. this trial are encouraging and there could the Industry Services team, particularly the be merit in follow-up trials, the Institute advises ongoing investigation into the technical > Approximately 75% of the total guaiacol people who are considering the use of this performance of wine closures, and more present was apparently extracted from technology commercially to conduct their recently the issues relating to the potential grape skins when they were homogenised own trials, and to undertake rigorous sensory spoilage yeast Dekkera/Brettanomyces. in a 10% ethanol solution. evaluation on samples pre- and post-treatment, Investigations into the effect of bushfire and to have the samples independently smoke on grapes and wine also resulted in > A small amount of guaiacol and 4- analysed for guaiacol concentration. a number of media enquiries. The majority of methylguaiacol remained in the marc queries are answered either by telephone, after pressing. References email or facsimile, and Industry Services staff supply approximately 500 technical papers Conclusions Martin, J.T. (1960) Determination of the or other pieces of relevant literature to callers components of plant cuticles. J. Sci. Food each year, via the John Fornachon Memorial The Institute had previously been asked to Agr. 11, 635-640. Library. Increasingly, Industry Services staff consider if smoke resulting from controlled are also able to direct callers to web-based burning of bushland might result in a taint Radler, F. (1965) The main constituents of information, both on the Institute's own, occurring in grapes in nearby vineyards, the surface waxes of varieties and species and other web sites. Furthermore, the support and had provided advice that there was a of the genus Vitis. Am. J. Enol. Vitic. 16, facilities provided by other Institute possibility that such a taint could arise. 159-167. research and library staff are important in The results of this investigation provide conclusive evidence that tainting of grapes Table 8. Enquiries received by Industry Services advisory staff during the past three years* and wine by bushfire smoke can occur, and potentially have a major economic impact. 2000/01 2001/02 2002/03 Many affected white juices, wines and especially sparkling base wines, were deemed to be ‘unfit for purpose’ and were Wineries 1155 1008 1184 consequently severely downgraded in terms Government organisations 117 101 97 of value. It remains to be seen what the commercial impact of the elevated guaiacol Other 359 469 368 levels in many red wines will be. Students 26 31 27

As Australian viticulture continues to spread Total 1657 1609 1676 into locations that might be considered as more bushfire-prone than many established

*This does not include the calls to other senior Institute staff, e.g. the Viticulturist responds to over 400 enquiries and the Health and Regulatory Information Manager responds to over 300 enquiries each year. 39 40 Industry Services Teams’ report years. However, itisevidentthatthenumber and assuchthegrowth hasslowedinrecent occurred inthewiderAustralianwineindustry, the increase reflects theexpansionthat has year isshowninFigure 12.To somedegree, advisory servicebetween1992andthe current The numberofenquirieshandled by the (see Table 13),asdiscussedabove. technical informationreceived from journalists 2002, asubstantialnumberofrequests for from industrysuppliers,andasreported in 'other' categoryinTable 8are received largely and wines.Therequests recorded in the to theeffects ofbushfire smokeongrapes first halfof2003,including43callsrelating record of980callswasrecorded duringthe highest everrecorded at71%.Asix-monthly requests received from wineries wasthe to theprevious year. Theproportion of the numberofenquiriesreceived compared Table 8.Thefigures showa4.2%increase in Ruediger during2002/2003ispresented in Ella Robinson,Geoff CoweyandGreg Peter Godden,AdrianCoulter, MarkGishen, A summaryoftheenquiriesreceived by is avaluableresource tothewiderindustry. MemorialLibrary)andinformal, Fornachon store ofinformation,bothformal(intheJohn wine research andpoliticalbodies.Thevast having linkstoAustralianandinternational wine industrysuppliersandjournalists, service, increasingly bodies, forGovernment The Institutealsooftenactsasareferral of investigation. technical references relating tothearea are oftenaccompaniedbyanumberof to prevent themre-occurring. Thereports encountered, andprovide adviceonhow the underlyingcausesofproblems written inawaywhichseekstoexplain the problem beinginvestigatedandare amount oftechnicalinformationrelating to for theclient.Thesereports containalarge result inafullwrittenreport beingprepared 7 Most oftheinvestigationsrecorded inTable in responding tomanyoftheseenquiries. Services Laboratoryplaysanimportantrole theanalyticalcapacityofIndustry and supplying relevant informationtocallers,

Number of enquiries 1000 1200 1400 1600 1800 200 400 600 800 0 9219 9419 9619 9819 0020 022003 2002 2001 2000 1999 1998 1997 1996 1995 1994 1993 1992 Figure 12.Enquiriesreceived byadvisorystaff, 1992to2003 conventional yeasts. stuck fermentation,andtheuseof non- vineyard effects andeffect onwine, phenolics, salinitybothintermsof soiland microbiological stabilityissues,red wine Dekkera/Brettanomyces discussion onrecent Roadshowshavebeen 8. Themostcommonareas ofinformal the numberofenquiriesrecorded inTable would accountforasubstantialincrease in this contactwere formallyrecorded thenit informal advice,anditisconsidered thatif an importantvehicleforthedeliveryof presentations, Roadshowsare alsoconsidered audience. Inadditiontotheformal tailored totheinterests andneedsofthe activity, inorder thatseminarsare closely approximately 40areas ofcurrent Institute be madeateachseminarfrom alistof are askedtoselectthepresentations to Regional winemakers’andgrowers’ associations (see Appendix1). winemaking and tenseminarsfive andTasmania),South Australia,Victoria states were visited(Western Australia, During thecurrent reporting periodfour seminars were presented in18regions. end ofthe2002Roadshow‘season’, day seminar. Duringthe24monthsto making twelvepresentations ineach full- with sixseniorInstitutestaff members from alloftheInstitute’s research teams, these events,theyrely heavilyoninput are responsible fortheorganisationof Whilst theIndustryServicesstaff members in thesecondhalfofcalendaryear. local vignerons’ associations,andare held generally organisedinconjunctionwith wine growing regions. Roadshowsare which are madeonarotating basisto24 Roadshow seminarandworkshoptours, Advisory Servicesare supportedby The ConsultationandInvestigative important source oftechnicalinformation. wine industryregard theInstituteasan a largenumberofpersonnelintheAustralian an historicallyhighlevel,andindicatesthat of enquirieshasstabilisedtosomeextentat workshops were presented and other Trouble free Institute’s members are alsoregular contributorstothe for theConference. IndustryServices team the stagingofapproximately 70workshops has primaryresponsibility forcoordinating Planning andProgram Committees, and inJuly2004,servingonboththe Melbourne Technical Conference, whichwillbeheldin Industry planning ofthe12thAustralianWine Manager hadsubstantialinvolvementinthe During thereporting period,theteam showsare encouraged. other wine Similar approaches Course. from the in successful recent most participants the from Showandothershowswere selected Wine some AssociateJudgesforthe2002Adelaide participants andguestjudges.Asinthepast, the coursewasreceived from anumberof Extremely positivewrittenfeedbackregarding judging aspartofawineshowpanel. simulate thedynamicsassociatedwith discussion exercises thatwere intendedto included anumberofnewtastingand enhanced, andthesixteenthcourse The coursecontentiscontinuallybeing was changedfrom theprevious three days. increased substantiallysincetheformat and demandforplacesinthecoursehas course hasproved tobeverysuccessful, presentation ofthecourse.Thefour-day andwinemakersassistedinthe journalists four days,and14leadingwineshowjudges, includes over40hoursofactivitiesthe presented usingafour-day format,which performance. Thiswasthefourthcourse develop andtesttheirsensoryevaluation thirty participantstheopportunityto Course washeldinJuly2002,givinganother Assessment The sixteenthAdvancedWine of itsfunctionswill beincorporatedinto project ceasedon30June2003,andmany companies andindustrysuppliers. This on afeeforservicebasisvarious wine protocols whichrelate totrials beingconducted Analytical Serviceonthedevelopment of staff alsoprovide advicetotheInstitute's marketed totheindustry. Industry Services products andtheevaluationofnewmaterials relative performanceofcommercially available The evaluationstakeoneoftwoforms:the winemaking processing aidsandadditives. wine analysis,andtheevaluationof development andevaluationofmethods GWRDC-funded project fortheimprovement, The IndustryServicesLaboratorymaintainsa Peter Valente Cowey, Greg Ruediger, Trudy Wallis and Dr AlanPollnitz,DimitraCapone,Geoff Gishen, EllaRobinson,DrMarkSefton, Staff: processing aidsforwinemaking Evaluation ofnewanalyticaltechniquesand 2), duringthecurrent reporting year. University’s Wodonga Campus(seeAppendix other lectures tostudentsfrom LaTrobe students attheUniversityofAdelaide,and provided elevenhoursoflectures toOenology and conferences (seeAppendix1),and also provide presentations seminars forexternal Peter Godden,AdrianCoulter, Mark Technical Review , periodically a new project Application of research manufactured from synthetic materials, five closures tested. In particular, wine and development to industry problems three of which are produced by extrusion sealed with the ROTE closure continues to and opportunities. The first subject to processes (ECORC, Nomacorc and NuKorc) retain the greatest concentration of SO2, be addressed by the new project will be and six by moulding (Aegis, Auscork, but the concentration was not significantly Investigations into the relationship between Betacorque, Supremecorq, Californian different to that in wine sealed with the Dekkera/Brettanomyces yeast and wine ‘Tage’1 and Integra). Various aspects of the Altec closure. Wine sealed with the Altec spoilage, thus during the current reporting performance of the closures are being closure demonstrated a slightly lower level year, many of the resources of this project examined, including those relating to each of brown colour development than wine were directed toward preliminary research closure's physical characteristics and extraction sealed with the ROTE closure at the four- and methods development associated with from the bottle, chemical analysis of the year post-bottling time point, as measured the Dekkera/Brettanomyces investigations. wine in order to examine apparent sealing by the conventional cuvette method, but performance, and sensory analysis. this was reversed in readings made using As previously reported, during the preceding the in-bottle test. However, it is clear that three years the vast majority of resources The retention of SO2 in wine sealed with each differences among readings made using allocated to this project were used for the of the closures continues to be a pivotal the two methods are minimal, and in

Closure Trial, a wide-ranging trial that is measurement in this investigation. Loss of SO2, general the measurement of brown colour examining the technical performance of 14 presumably due to the degree of oxygen by the non-destructive test (Figure 13) different closure types. Testing in this ingress allowed by the closure, has been matches the cuvette measurements project continues, albeit at a lower level of intensity than in previous years, and results Table 9. Mean SO2 concentration and optical density at 420 nanometers (OD420) following 48 months of storage in an inverted position of the trial, particularly those associated c with the measurement of sulfur dioxide (SO2) 48 months OD420 (cuvette measurements) (au) concentration and wine browning as measured inverted storage (n = 20)a a by optical density at 420 nanometers (OD420), (n = 20) and sensory evaluation of wine sealed with each of the closures, continue to be b Altec Free 14 (2) 0.156 (0.003) published periodically. During the current Total 79 (3) year results were published in Technical Review issue #1391 (August 2002), and the One plus One Free 9 (2) 0.184 (0.007) Proceedings of the Romeo Bragato Total 70 (3) Conference1, Christchurch, New Zealand, September 2002. The trial continues to Reference 2, 44 mm cork Free 6 (4) 0.194 (0.018) attract interest from both industry and Total 62 (11) mainstream media (see Table 13), and in a Reference 3, 38 mm cork Free 5 (4) 0.205 (0.016) letter to the Institute from a representative Total 56 (12) of a leading UK supermarket chain the trial was described as a "catalyst for the changes ROTE Free 15 (4) 0.163 (0.011) in closure use that are now apparent". Total 81 (5)

Note: The ‘closure trial’ was not designed an = 18 for ROTE, bfigures in parentheses are standard deviations, cau = absorbance units to be a survey of closures available in the marketplace, and not all of the closures found to correlate well with the development extremely closely for the five closures available at the time the trial commenced of brown colour in the wine, and also the tested at the four-year post bottling time were included. Readers should also note that rating of oxidised character during sensory point (for further documentation of this the currently available stocks of some of the evaluation, at all testing intervals. The measurement, see Institute publication #731). closures examined might differ from those concentrations of free SO2 recorded in It is equally apparent that the separation of available when the trial was implemented, wine sealed with each of the closures at six closures on the basis of brown colour in and that the results reported represent the months, were also found to be excellent the wine as measured by both tests, is performance of each closure when used to predictors of free SO2 concentration and closely related to the concentration of SO2 seal only one wine, under the conditions browning and rating of oxidised aroma in retained in wine sealed with each of the defined. Care should therefore be exercised the wine after 24 months, and to a lesser closures at 48-months post bottling. in relating the results to other wine types, extent after 36 months. The fourth Additionally, it is clear that small or to wines bottled and stored under different anniversary of bottling the trial wine differences in SO2 concentration identified conditions. It should also be noted that occurred late in the reporting period, and as early as the six-month post bottling time some closures may be suitable for use in at this point wine sealed with only five point, relate strongly to increasingly large products with a short shelf life, and may closures: the ‘ROTE’ roll-on tamper-evident differences in brown colour in wine sealed perform well over a particular time span, ‘screw cap’, the Altec and One plus One with all of the closures, as the trial progresses. but less well over a longer term. Winemakers technical corks, and the reference 2 and therefore need to assess the anticipated reference 3 corks, were tested for SO2 shelf life of the wine to be bottled when concentration, OD420 using conventional making decisions on closure use. cuvette measurements (Table 9), and sensory analysis for aroma only. Additionally, wine The closures that are being examined in this sealed with all 14 closures was tested for study comprise a roll-on tamper-evident OD420 using a non-destructive in-bottle (ROTE) screw-cap closure, two grades of measurement developed by Skouroumounis et. natural conventional cork (reference 2, 44 al. (Figure 13) (Institute publication #731). mm length and reference 3, 38 mm length), two ‘technical cork’ closures (cork-based Data presented in Table 9 indicate that the closures that also contain a synthetic important trends observed during earlier component: Sabaté ‘Altec’ and Amorim testing have continued up to the four-year One plus One ‘Twintop’), and nine closures post-bottling time point, with regard to the

1 The Institute wishes to make it clear that any reference to ‘Tage’ closures in these publications, and also in this publication, are references to closures that were supplied by Esvin report Teams’ Services Industry Wine Resources, Auckland, New Zealand in May 1999, and not to closures supplied by Novembal. The Institute has been informed that the closures being examined were made by a Californian manufacturer, and not by Novembal, and Novembal has informed the Institute that these closures are not the same as Tage closures now being made by Novembal. The Institute takes no position on Esvin’s right, or that of the Californian manufacturer, to make or supply closures under the name ‘Tage’. 41 42 Industry Services Teams’ report loss byabsorption. degradation couldbedistinguished from that thelossofwinecomponentsby chemical in all-glasscontainers(sealedampoules), so control samplesstored withandwithout air study. Inthiscaseitwasalsoessentialtohave practical andwastheapproach takeninthis technical reasons, thelatteroptionisfarmore compounds inwineduringstorage.For the decrease inconcentrationofthese compounds, or, indirectly, bydetermining and analysingtheextractsforabsorbed directly byextractingtheclosures themselves determined inoneoftwoways—either Absorption offlavourcompoundscanbe 144, June2003). this wasgivenin synthetic closures (apreliminary report on 2 and3),atechnicalcorkclosure, andseven two gradesofnaturalbarkcork(Reference a roll-on tamper-evident screw cap(ROTE), The closures usedinthis‘scalping’trial were publications #666,671,675,676,and677). oxidation andsensoryproperties (Institute May 1999)whichfocussedonwhitewine Institute’s earlierclosure trial(bottlingin were from thesamebatchesusedin closures. Theclosures usedinthistrial (flavour scalping)ofvariouswinebottle others) tostudytheabsorptivecapacity (by DrMarkSefton,DimitraCaponeand A longtermtrialwassetupseveralyearsago publication #702). box’ packagingmaterials(Institute wine wasalsodemonstratedfor‘bagin to absorbaroma activecompounds from improvement inwinequality. Thiscapacity phenomenon clearlyresulted inan #616). Inthisinstance,theabsorption during bottlestorage(Institutepublication closures absorbingtheTeCA from thewine ‘decontaminated’ asaresult ofthecork during barrel maturation,were effectively compound 2,3,4,6-tetrachloroanisole (TeCA) contaminated withthemusty-smelling observed acasewhere red wines, closures duringbottlestorage.Recentlywe therefore, alsotakeplacewithwinebottle as ‘flavourscalping’.Suchabsorptioncould, the product, aprocess sometimesknown quality byabsorbingaroma compoundsfrom other foodproducts canalsomodifyproduct well knownthatpackagingcomponentsin in termsoftheseprocesses. However, itis aroma andflavourisgenerallythoughtof The impactofwinebottleclosures onwine on winearoma andflavour. be extractedintowinewithvariouseffects that otherconstituentsofclosures canalso imparted towine.There issomeevidence whereby anunwantedmustyaroma canbe closures intowineiswellknownasameans 2,4,6-trichloroanisole (TCA)from corkbark of oxygen.Theoccasionalextraction by actingasapartialbarriertothetransmission changes duringbottlestorage,principally closures playarole inthewaywineflavour It haslongbeenrecognised thatwinebottle Flavour scalpingsub-project Technical Review issue also affected bytheclosures (otherthan one winetoanother. Someesterswere and theircorresponding acids)varies from (relative startingconcentrationsofesters because thefactorsaffecting suchchanges be extrapolatedfrom thiswinetoothers, others decreased. Thesechangescannot esters increased inconcentrationwhile esterification reactions inthewine. Some a result ofcompetinghydrolysis and esters changedinthecontrol winesas The concentrationofthesimplefermentation and are notaffected bybottleclosures. (including control winesstored underair) therefore chemicallystableinwine year period.Allofthesecompoundsare the controls orbottledwinesoverthetwo changed concentrationsignificantlyineither lactone, 4-ethylphenoland4-ethylguaiacol methylguaiacol, vanillin, oak-related componentsguaiacol,4- Neither isobutylmethoxypyrazinenorthe control samplessealedinglassampoules. decreases inconcentrationwasobservedfor decreased duringthestorageperiod,similar Where theconcentrationofcompoundshad from thewinesealedunderROTEclosure. absorption ofanytheflavourcompounds In nocasewasthere anyevidenceof was sealed. which tookplaceregardless ofhowthewine the closures, otherstochemicalmodification changes canbeattributedtoabsorptionby wine changedsignificantly. Someofthese all oftheflavourcompoundsaddedto storage, theconcentrationofsome,butnot storage facility. Aftertwoyearsofbottle inverted positioninaconstanttemperature the closures listedaboveandkeptinan was thenbottled,thebottlessealedwith compounds wewishedtostudy. Thewine wine wasspikedwitheachofthevolatile a rangeofflavourcompounds,Semillon To observetheeffects ofbottleclosures on

OD420 (au, 10 mm cuvette equivalent) for ref 3,andOneplus Oneandn=8forref 2from 42months onwards) inverted position,asmeasured byopticaldensityat420nm (OD Figure 13.In-bottle, meandevelopmentofbrown colourinwinesealedwitheachclosure type,stored inan 0.10 0.12 0.14 0.16 0.18 0.20 0.22 0.24 0.26 0.28 0.30 21 43 64 8061 82 03 248 42 36 30 24 18 12 6 0 48 42 36 30 24 18 12 6 0 Nomacorc Supremecorq Ref3,38mm Ref2,44mm Oneplus NuKorc Integra Auscork Aegis Nomacorc Supremecorq mm Ref 3,38 mm Ref 2,44 One Plus NuKorc Integra Auscork Aegis cis- and trans-

oak Storage time(months) 15% to45%. closures only. Thisabsorptionrangedfrom was partiallyabsorbedbythesynthetic ‘lychee’ charactertosomewhitewines, The monoterpenerose oxide,whichgivesa did winesstored undernitrogen. stored underairhad30%lesslinalool than some effect ofheadspace.Control wines two years.Inthecontrol samplesthere was 10% oftheaddedlinaloolremained after 5% oftheaddednerol andgeraniol and terpin hydrate.Inthesetrialwines,lessthan relatively weak-smelling alphaterpineoland convertedtothe to linaloolwhichisinturn pH, nerol andgeraniolare convertedmainly wines waschemicaldegradation.Atwine concentration ofthesecompoundsinthe The overwhelmingfactoraffecting the monoterpenes linalool,geraniolandnerol. closure typeontheconcentrationof There appeared tobelittleeffect ofthe to 70%(oneclosure). ethyl decanoatefrom 50%(twoclosures) (two closures) to50%(oneclosure), and Ethyl octanoateabsorptionvariedfrom 30% three closures) to15%(intwoclosures). absorption wasslight,rangingfrom 5%(in these esters.Forethylhexanoatethe showed significantlygreater absorptionof and 35%respectively). Thesyntheticclosures more oftheselasttwocompounds(10% The technicalclosure absorbedslightly ethyl octanoateand20%ofdecanoate. corks) absorbednoethylhexanoate,5%of Both naturalclosure types(Reference 2and3 increasing esterchainlength. amount ofabsorptionincreased with the closures, andforeachclosure type,the decanoate were allpartiallyabsorbedby Ethyl hexanoate,ethyloctanoateand isovalerate were unaffected bytheclosures. ethyl isobutyrate,butyrateand ROTE) tovaryingdegrees. Theshortchain 420

). (n=12forallclosures to36months,n=9 ROTE Altec Californian'Tage' ECORC Betacorque ROTE Altec ‘Tage’Californian ECORC Betacorque The taint compound naphthalene (added to fining agents examined were bentonite, PVPP, worked with Dimitra Capone under project give an idea of the absorption of structurally egg albumin, casein, gelatin, isinglass, AWR 6, to develop a GC-MS method for the similar compounds) was substantially absorbed, activated carbon (charcoal) and copper sulfate. determination of the concentration of both particularly by the synthetic closures. The 3 methylbutyric, and 2-methylbutyric acids least absorptive of these removed 75%, Even the mildest fining removed more than in wine. while 90% was removed by the most 20% of every ester analysed. Generally, the absorptive. The natural and technical corks shorter chain, more polar, more volatile Also in response to the requirements of absorbed less of this compound (30% and esters were more affected by the fining agents, the Dekkera/Brettanomyces investigations, 40%) respectively. presumably due to their stronger hydrogen development of HPLC methods for the bonding, a mode of action attributed to some quantification of coumaric and ferulic acids The grape derived hydrocarbon known as of these fining agents. This trend is the opposite in both red and white wines and grape TDN can give a kerosene-like flavour to some to that observed in the 'flavour scalping' trial homogenates has continued. Coumaric white wines. It can sometimes be found in reported above where it is likely that the and ferulic acids are precursors for the relatively high concentrations in Riesling hydrophobicity of the longer chain esters makes production of the spoilage compounds 4- wines, particularly those that have been in them more prone to adsorption out of the wine. ethylphenol and 4-ethylguaicol by Dekkera/ bottle for two years or more. This compound Of the monoterpene alcohols, linalool, nerol, Brettanomyces yeast, and these methods was the most strongly affected by the geraniol and alpha-terpineol, only linalool are considered crucial for the progression of closures. All of the synthetic closures had was significantly affected by fining by agents the Dekkera/Brettanomyces investigations, removed between 96% and 98% of the other than carbon, which removed approx. allowing the development and subsequent TDN after two years. The natural corks and 10% of all four compounds. The loss of degradation of the precursors to be technical closures removed 50% and 70% linalool ranged from 5% due to isinglass measured during fruit ripening, winemaking respectively, while the ROTE closures removed up to 18% due to PVPP. and subsequent wine storage. none at all. TDN is sometimes regarded as undesirable when present in high quantities, Damascenone and beta-ionone were Further methods that are likely to greatly so the capacity of closures to remove this unaffected by fining, except by bentonite and enhance the Institute’s investigative particular flavour compound (and perhaps copper sulfate which removed about 5%, capabilities in future years, have also been others) from wine is certainly not always PVPP which removed 10%, and carbon which developed during the current reporting deleterious to wine quality. removed almost 20%, of both compounds. period by the Institute’s Mass Spectrometry The oak compounds were little affected by facility manager Mr Yoji Hayasaka and In summary, relatively non-polar volatile fining. The largest effects observed were technical officer Ms Gayle Baldock, and are compounds can be absorbed by synthetic from carbon, which removed only up to discussed elsewhere in this report. and natural cork closures, but not by ROTE 9% of the oak compounds. screwcaps. Synthetic closures showed a much Assistance and advice were provided to the greater capacity to absorb these non-polar Naphthalene and TDN behaved in a similar Analytical Service by reviewing method compounds than did natural cork bark or manner to each other, with about 10% validation reports for conformance with the technical closures. The technical closure more TDN being removed (compared to requirements of the Institute's quality system, behaved in a similar fashion to the natural naphthalene) in every case. The extent which is based on NATA requirements and corks but had a slightly higher capacity for of removal of TDN ranged from 30% for ISO Guide 25. This work is conducted by absorption. Among the synthetic closures, isinglass up to 70% for carbon. the Institute's Quality Liaison Manager, Mark some consistently appeared to have had a Gishen, who is part of the Industry Services higher capacity for absorption than others. Fining has the potential to affect the aroma team, and the following methods were Some compounds (oak components, and flavour profile of wine. The choice of reviewed during the year: Tetraconazole and isobutylmethoxypyrazine, 4-ethylphenol, TDN) fining agent is important, as not all compounds cyproconazole in wine, juice, grapes and marc are completely stable in wine, even with an air are affected in the same way. Note that by GC-MS; Quinoxyfen in wine, juice, headspace, while others (monoterpenes, the Institute wishes to reiterate that fining grapes and marc by GC-MS; Histamine and damascenone) are substantially degraded in is a very important part of winemaking and tyramine in red and white wine by HPLC; wine whatever the closure. that the inevitable scalping that takes place Fludioxonil, pyrimethanil, fenhexamid, during fining should not be interpreted as tebufenozide and cyprodinil in juice wine Further data analyses and interpretation an argument for doing away with fining. and grapes by HPLC; Isovaleric acid in red of the data from this study are currently and white wine by GC-MS; Emamectin in underway. At this point in time, it appears Analytical method development and evaluation juice, wine, grapes and marc by HPLC. In clear that the choice of closure may have a addition, NATA accreditation was obtained dramatic effect on wine sensory The majority of the methods development work for the methods: Malathion and Fenitrothion characteristics not only due to gross that has been performed during the current in wine and juice. oxidative changes and transfer of taint year relates to the investigations into the compounds described earlier (e.g. Institute relationships between Dekkera/Brettanomyces Targeted training of wine industry personnel: publication #666) but also through direct spoilage yeast and wine. compilation of a technical reference manual and absorption of compounds to the closure delivery of complementary workshops. material. As closure manufacturing The development of a GC-MS method for the technologies and materials are likely to change quantification of isovaleric acid (3-methyl Staff: Peter Godden, Adrian Coulter, Mark due to ongoing commercial developments, butyric acid) in wine, using a deuterated Gishen, Ella Robinson, Dr Mark Sefton, winemakers must at all times conduct their standard of isovaleric acid that was produced Dr Alan Pollnitz, Dimitra Capone, Geoff own trials to make informed choices about at the Institute under the GWRDC-funded Cowey, Greg Ruediger, Trudy Wallis and the closures most suited to their preferred project AWR 6 Grape Composition and Wine Peter Valente styles of wine and commercial objectives. Flavour, was reported in the previous Annual Report. Isovaleric acid is reported in the The primary aim of this project is to Flavour absorption by fining agents literature as an important spoilage compound produce flexible and updateable information produced by Dekkera/Brettanomyces yeast. packages on selected technical subjects, An Honours student, Matthew Clarke, under the During the current year it was ascertained which will be delivered to wine grape levy supervision of Dr Alan Pollnitz, has recently that during GC-MS analysis of isovaleric payers via the internet, and workshops to completed a project on the effect of fining acid, a second acid, 2-methylbutyric acid, be held in all major grape growing areas of treatments on the concentration of aroma was eluting at the same time. Subsequently, Australia in conjunction with the existing compounds in a Chardonnay wine. The Geoff Cowey of the Industry Services team Roadshow seminars. report Teams’ Services Industry 43 The Institute has a vast amount of collective New material is periodically added to the site, that a change in one variable, whether knowledge pertaining to grape and wine and the existing information is enhanced. intended or not, can have a potentially production, much of it generated by over During the current year, in conjunction with dramatic flow-on effect leading to the 48 years of research and through Industry the Communications and Publicity Manager, occurrence of an instability. Services' projects at the Institute, that have all the material on the ‘hazes and deposits’ been supported by Australian winemakers section of the site has been edited and in Such a winemaking change may be as and grapegrowers. Although this research some cases re-formatted, in order to enhance simple as picking fruit one degree Baumé has produced in excess of 730 technical its functionality. New digital images of deposits riper than in previous years. The resulting publications, there is also a great deal of isolated from wine have also been added. higher alcohol concentration in the wines, information generated and recorded in a more all other things being equal, may lead to informal manner by the staff concerned. In As discussed elsewhere in this report the fermentation problems with resultant addition, a great deal of data pertaining to combined and related areas of hazes and increase in residual sugar concentrations. the composition of Australian wine is stored deposits and microbiological instabilities These wines may take longer to complete in various databases at the Institute, some continue to represent a significant proportion malolactic fermentation (MLF) due to the of which have been in existence for several of the problem wine sample investigations elevated alcohol concentration, allowing decades. Collectively, this information is a conducted by the Industry Services team. the proliferation of unwanted microflora. resource of great potential value to the In many cases it is apparent that the problems Pre- or post-MLF growth of unwanted Australian wine industry. This project, could have been avoided if winemakers microflora, including Dekkera/Brettanomyces therefore, seeks to make a record of this were more aware of the causes of many as discussed below, will be favoured by the information, so that it can be delivered to types of instability, and were able to access availability of residual sugars, and such

18 16 2500 17 0.14 15 16 2000 0.12 15 14 0.10 14 1500 13 13 0.08 Count 12 1000 0.06 12

11 per Bar Proportion 0.04

Alcohol content (% v/v) 10 Alcohol content (% v/v) 11 500 9 0.02

8 10 0 0.00 19841990 1996 2002 1984 1986 1988 1990 1992 1994 1996 1998 2000 2002 1984 1986 1988 1990 1992 1994 1996 1998 2000 2002 Year Year Year

Figure 14. Median (left panel) and mean (centre panel) ethanol content of Australian red wines by vintage, 1984 to 2002, and numbers of samples analysed (right panel) (Source: The Australian Wine Research Institute Analytical Service database)

the industry in a manner in which it is both practical and relevant tools with which growth may directly or indirectly lead to

readily useable, and relevant to those involved to trouble-shoot the problems, before the binding of free SO2. This loss of SO2 in day-to-day wine production. It is envisaged significant wine spoilage had occurred. may in turn leave the wine more prone to that the project will be ongoing, and that The website and workshops address these oxidation and further microbiological activity, many areas of interest to winery technical issues, providing practical trouble-shooting which in turn may lead to the formation of personnel will be addressed in due course. information and simple diagnostic tests, a deposit in the wine. The Industry Services with which to isolate and identify a wide staff considers that this scenario is becoming The information collated to date has been range of hazes and deposits in wine. more common, and is apparent in both in posted to the Institute's web site over the requests for technical assistance received past two years by Industry Services staff. During the reporting year, five Trouble-free from industry, and in the nature of the Australian winemakers and grapegrowers winemaking – identification and management problem solving investigations conducted. can access the information at of common wine instabilities workshops Increasingly, data contained in the Institute’s http://www.awri.com.au/practical_solutions/ were staged under the auspices of this various databases is being collated under the (or alternatively, visit the Institute’s website project, in three states (see Appendix 1). auspices of this project in order to enhance at www.awri.com.au, and select 'Practical The content of this workshop was further the content of the workshops, and to monitor Solutions' followed by 'Hazes and Deposits'). enhanced during the year in order to focus various compositional trends in Australian This section of the site is password protected more strongly on helping winemakers wine. It is apparent that these data may (the password has been previously supplied understand the causes of common wine be used to explain the number of wines to Australian wineries). Australian grape instabilities. In addition to discussing investigated that exhibit the type of instability growers and winemakers who require the options for formal quality management problem discussed above. As an example, password and who can confirm that they systems, the workshops contain practical Figure 14’s left and centre panels plot the pay the Wine Grapes or Wine Research advice on the most efficient way to median and mean ethanol concentrations Levy, can obtain the password from the incorporate routine analytical testing into of Australian red wines recorded in the Institute's Librarian (Catherine Daniel on quality management programs, and commercial Analytical Service database, by email: [email protected]). suggestions on the most efficient way to vintage between 1984 and 2002. Figure However, the 'Industry Development and use the information generated to identify 14’s right panel provides the numbers of Support' section of the Institute's website where problems exist, or to identify samples from each vintage analysed. Whilst also contains a great deal of other potential problems before they occur. A these data represent a cross section of technical information, which is readily recurring aim and theme of the workshop Australian wines by value, they are probably accessible by interested parties. is to foster an understanding of how wine skewed towards relatively higher rather than

Industry Services Teams’ report Teams’ Services Industry compositional variables are linked, such lower value wines. The data demonstrate a 44 steady increase in ethanol content over this Information and document delivery services Specialised information services period, from a mean of approximately 12.6% in 1983, to a mean of approximately 14.2% The Library has excellent access to international The Library staff continue to be actively in wines from the 2002 vintage. Interestingly, databases, particularly in the fields of science, involved in the production of specialised the rate of increase in mean ethanol technology and medicine. If requested, the information products for the benefit of concentration appears to have accelerated Librarian, Catherine Daniel, will carry out the wine industry, such as the annual and in 2001 and 2002, and in 2002 the median online searches on commercial databases web-based editions of the Agrochemicals upper quartile of ethanol concentrations on any appropriate topic (on a fee-for- registered for use in Australian viticulture, comprises wines that contain between service basis). Alternatively, Library staff the bimonthly Technical Review, and several approximately 14.9% and 16.5% ethanol. can provide, free of charge, a report of in-house technical information databases. By consulting Figure 14’s right hand panel, it relevant articles indexed on the Library’s can be ascertained that this quartile contains in-house databases. Library collection approximately 400 individual wines. It appears plausible that trends such as these The Library continues to enjoy strong A total of 74 monographs and 20 conference are contributing to the number of Australian support for all its services with increases proceedings and over 3,600 new records were wines in which substantial potential quality in requests over that serviced in added to the library databases during the could be lost during processing. This 2001/2002 (Table 10). During the year, year. Further to this, 9,011 records sourced quality loss may be manifested in a the Library serviced a 24% increase in from the International Food Information diverse range of symptoms ranging from information requests from industry Service (IFIS) were also added to the library incomplete primary fermentation to personnel and a 19% increase in requests databases during this period. Over 70% of subsequent microbiological instabilities from Institute staff. The number of these fully abstracted records replaced existing causing deterioration of the sensory Institute publications forwarded rose library database records where only the characteristics of wine, and the formation by 24% over the previous year’s 33% citation was available, thus enhancing the of hazes and deposits, as described above. increase, while requests for articles from usefulness of information the library indexes The continued and enhanced targeting of Technical Review rose by 19% over the on its databases. Industry Services activities to foster industry previous year. The Library staff further wide understanding of these types of issues handled a significant increase in requests Following a rigorous review of the library's is therefore considered crucial in preventing for interlibrary loans from the Library’s in-house database, the total number of quality loss, with consequent substantial collection during the year. Industry’s records indexed in the database fell slightly, cost savings for the Australian wine industry. increased awareness and use of the although the number of unique records has Of particular interest in this respect is the Institute’s web-accessible database has in fact increased. finding by Helmut Guth and colleagues that also contributed to an increase in high alcohol concentrations can attenuate enquiries, and this important service The Library subscribes to 54 journals and the impact of volatile wine aroma compounds continues to be a valuable way for receives approximately 70 annual reports, significantly (Guth and Sies 2002). Ironically industry to access its technical literature, journals and newsletters through exchange therefore, the quest to accumulate high 24 hours per day, 7 days per week. and donation. The Library also maintains a concentrations of aroma and flavour collection of over 22,000 reprints. compounds in wine by getting very ripe Document delivery fruit could well be counteracted by the parallel increase in ethanol content of the The Library can supply either books or final wine. Clearly the industry is cognisant photocopies from its collection or obtain such of this and therefore appear very supportive items for wine industry clients through the of the Institute’s extremely challenging quest interlibrary system. Patents or standards can to seek ‘low alcohol yeast’ (see for example also be ordered. Electronic ordering and Institute publication #684). delivery services mean that most interlibrary requests are fulfilled within five days. Charges Reference: apply for the supply of some items.

Guth, H.; Sies, A. (2002) Flavour of wines: Table 10. Summary of information requests during 2002/2003 towards an understanding by reconstitution experiments and an analysis of ethanol’s Wine industry Staff Other6 Total % change effect on odour activity of key compounds. Blair, R.J.; Williams, P.J.; Høj, P.B. (eds.) 2003 2002 2003 2002 2003 2002 2003 2002 2003 Proceedings of the Eleventh Australian Wine Industry Technical Conference 7-11 October Information requests 1081 869 943 792 1584 1702 3608 3363 7% 2001. Adelaide, South Australia. 128-139. Interlibrary loans: > requests sent1 29 182 505 345 534 527 1% Provision of technical information 2 > requests received 88 53 66% 3 Staff: Rae Blair, Catherine Daniel, Ingrid Technical Review requests 230 193 19% Oats, Melissa Francis Technical Review articles forwarded4 1179 970 14% The John Fornachon Memorial Library holds Articles forwarded5 1424 1031 28% the largest collection of wine technical Number of Institute literature in Australia. The Library’s principal publications forwarded 725 585 24% responsibility is to provide technical information to the Australian wine industry Articles photocopied and to the researchers of the Institute. The in JFML 3874 3740 4% Library is also used extensively by other groups such as students, institutional researchers, 1 Staff at the JFML sent a request to another library for an article. government bodies and private companies 2 Requests received by the JFML from other libraries for articles from our collection. (see Table 10 for requests for information 3 Number of requests received for articles published in the Technical Review. 4 Number of articles forwarded (usually more than one article is requested). serviced during the year). 5 Number of other articles forwarded, excluding staff publications (usually more than one article is requested). 6 90% of ‘other’ requests come from students and Government sources. 45 46 Industry Services Teams’ report Table 11.Numberofrecords ontheLibrary’s catalogue,informationandweb-accessibledatabases ntdSae fAeiaFdrlRgse 877 393 43,282 404 Web accessibledatabase United StatesofAmericaFederalRegister ISYS –fulltextretrieval database REGS: European Communitywinelegislation Library informationdatabases newsletters,statisticsandannualreports JOURNALS: journals, Theses, scientificandmedicalpapers AWRI_Database: Books,conference proceedings, Library cataloguedatabases on relevant conferences, regulatory the Institute’s research aswellupdates provides progress reports totheindustry on Australia andoverseas. organisations andindividuals,both in andother subscription, bygovernment wineries inAustraliaand,through Grapes Technical Review Technical Review is updatedonaregular basis. edition (http://www.awri.com.au/agrochemicals/) edition automatically, andthe web-based levy payersreceive aprintedcopyoftherevised registered foruseinAustralianviticulture the thirteentheditionof Dr SallyBellandCatherineDanielprepared As reported elsewhere inthisAnnualReport, Agrochemicals Grid web database,onanongoingbasis. database records topostontotheLibrary's via thedatabase.Librarystaff continuetoedit source oftheabstractsthatare accessible arevarious publisherswhosejournals the with copyrightapprovals obtainedfrom the restriction inaccess isenforced tocomply Industries ExciseLevyTax 1999.The and grapegrowers payingthePrimary toAustralianwinemakers via theinternet Libraryprovides accesstoitsdatabases The databases isgiveninTable 11. of theLibrary’s catalogueandinformation in-house databases.Asummaryofthesize records retrieved from anyoftheLibrary’s particular subjectsorauthors,listingthe The Librarianprovides reports, eitheron separate in-housedatabases. holdingsarejournal maintainedon wine legislationanddetailsofthelibrary's Library’s collectionoftheEuropean Union catalogue; thebibliographicdetailsof are indexedontheLibrary'sdatabase technical andmedicalreprint articleswhich books, conference proceedings, scientific, the Library'scollectionofover43,000 A singlesearch screen provides accessto Library databases and Wine Research Levypaying and Wine is received byallWine wt erhbeasrcs 25,406 (with searchable abstracts) Technical Review Agrochemicals covering . All 080 Correction topage6of 30/8/02 Suggestedwording for allergenlabellingfrom 14December2002 AWRI winstheMauriceO’SheaAward 2002 15/8/02 5/8/02 Correction bulletinre informationsentoutonThermovitinthe26/7/02 bulletin 29/7/02 Selectedaddresses were usedtosendouttheenglishtranslation ofanegative 26/7/02 NewwithholdingperiodforAvatar andLegend 26/7/02 2002/2003Agrochemical bookletavailabilityandDithaneDF 22/7/02 130 Lateseasonmanagementof 11/3/03 Managementofmouldyfruit 24/2/03 Press release regarding thenewGWRDC-funded project onallergiesand wine 26/9/02 Newregistration ofShirlanandCrop Care Captanforphomopsis 26/9/02 within thesixissues. interest publishedthroughout theyear literature abstractsandothermattersof current notes, technical fast accessto within theCDROMfacilitates mechanism simple A individuals. Levy paying charge toWine Gr aCDROManddistributedfree of via Technical Review The collectionofthe2002/2003issues is gratefullyacknowledged. Memorial Trust, andtheirongoingsupport contribution totheThomasWalter Hardy through regular generous financial support thepublicationof Hardy AOandhersonscontinueto technical_review/latest_issue/). DrBarbara (http://www.awri.com.au/ website Institute’s Technical Review each issue.Restrictedpassword accessto section viaarequest formavailablewithin articles featured inthe‘Current Literature’ Recipients of published technicalandscientificarticles. citation detailsandabstractsofrecently Review’s amendments andmedicalissues. CretLtrtr’scinprovides ‘Current Literature’ section article onAustralianwinespublishedinaNorwegiannewspaper20/7/02 and DithaneM-45information cane andleafblight,Mancozeb registration in Australianviticulture2002/2003 Technical Review has beenmadeavailable is alsoavailableonthe apes and Wine Research apes andWine Technical Review No. ofrecords Table 12.Emailbulletinssentduring2002/2003 may order search Agrochemicals registeredforuse Technical Botrytis Publishing Inc. K.F. Pocock,DrB.C.Rankine,Viticultural are gratefullyacknowledged: from thefollowingpersonsandorganisations tothelibrary Donations ofbooksorjournals Acknowledgements investment oftheEndowmentFund. together withtheincomegeneratedfrom Research andDevelopmentCorporation, annual grantfrom theGrapeandWine the Institute.TheLibraryisfundedbyan thefirstDirectorFornachon, ofResearch of from winemakersandfriendsofthelateJohn which wasestablishedin1969bydonations The InstituteactsastheTrustee ofthisfund, Endowment Fund The JohnFornachon MemorialLibrary during theyearandare showninTable 12. bulletins. Elevenemailbulletinswere issued should theywishtoreceive theemail email address ([email protected]) Australian wineindustryshouldsubmittheir bulletins, andinterested membersofthe addresses recorded toreceive theemail Australian wineindustry. There are 764email information tointerested membersofthe disseminating importanttechnical to beafastandcost-efficient wayof Technical IssuesBulletinservicecontinues The EmailAdviceandInformationon Email service Links between viticultural and oenological research

Staff: Dr Sally-Jean Bell, Professor Peter Høj, Kevin Pardon

A meeting was conducted with approximately 18 industry representatives in May to review recommendations on the use of agrochemicals. Thereafter, 11,000 copies of the Institute’s annual publication, Agrochemicals registered for use in Australian viticulture 2002/2003 were produced and the information duplicated on the Institute’s website. The recommendations were distributed to several wineries electronically to be reproduced in their own publications. The booklet was distributed with the Australian & New Zealand Grapegrower &

Winemaker, Technical Review and in the Melissa Francis Ingrid Oats Catherine Daniel Research to PracticeTM IPM and Spray Application manuals. The tables were also management on the phenolic composition High vigour VSP vines produced higher featured in Australian Viticulture. The of Shiraz grapes and wine. In the first season concentrations of seed catechin and 2002/2003 MRLs for Australian export markets there were four treatments consisting of epicatechin and skin anthocyanins than will be placed on the Institute’s website two canopy styles; Vertical Shoot Positioned high vigour SH vines as seen in the (http://www.awri.com.au/agrochemicals/mrls/). (VSP) and Scott Henry (SH) and two levels previous season. However, in contrast it of water status; high water (South end of was the low vigour SH vines that produced As a direct result of the Institute’s agrochemical rows) and low water (North end of rows). higher concentrations of seed catechin and publication and it’s Residue Analysis Service, The differences in water status were due to epicatechin and skin anthocyanins than Dr Sally Bell and Greg Ruediger devote a differences in soil depth and a decrease in low vigour VSP vines. As in the 2000/2001 significant amount of time in liaising with water pressure down the irrigation line season, the higher canopy density and major chemical companies. Companies are (South to North). The difference in growing lower bunch exposure exhibited by high increasingly aware of the importance the wine conditions resulted in measurable vigour vigour VSP vines in a mild season, appeared industry places on meeting export market differences. That is the vines at the North to be beneficial in terms of fruit phenolic specifications. Thus they are keen to work end of the rows exhibited low vigour while concentration despite the bigger berries. more closely with the wine industry and those at the South end exhibited high vigour. In contrast to the previous season the low the Institute to ensure that their products Results from 2000/2001 were reported in vigour SH vines with lower canopy density can be used in viticultural pest and disease the 2002 Annual Report. In the second and higher bunch exposure and small berries programs in such a way that the maximum season of this trial (2001/2002) a change increased fruit phenolic concentrations in residue levels set by Australian export in treatments occurred. The pressure comparison to low vigour VSP vines. markets will not be exceeded. differential in the drip line was corrected and consequently no difference in plant Sensory analysis of the 2001/2002 wines has During 2002/2003 Dr Sally Bell responded water status could be detected. Therefore, been completed. The following is a brief to 415 enquiries. The majority (68%) water stress was absent in 2002 but the summary prepared by Leigh Francis: Shiraz of enquiries were regarding the use of vigour difference was still apparent. These wines were made from grapes sourced from agrochemicals and the remaining 32% observations were confirmed by growth vines trained to either a Scott Henry or a related to general viticulture. measures in the 2001/2002 season. The Vertical Shoot Positioned trellis. In addition, 2002 season was also markedly milder a further treatment of high vigour versus Dr Sally Bell delivered a range of presentations than 2001, as illustrated by the Lyndoch low vigour was imposed. Paired at the Institute’s Roadshows in Tasmania, site Mean January Temperature (MJT). MJT comparison sensory difference tests for the Rutherglen (Vic), Bendigo (Vic), Pyrenees in 2001 was 28.2˚C in comparison to the attributes fruit aroma and overall astringency, (Vic), Yarra Valley (Vic), Mornington MJT in 2002 of 19.8˚C. were carried out. For the high vigour Peninsula (Vic), Swan Valley (WA), Great treatment no significant difference was Southern (WA) and Margaret River (WA) As seen in the previous season, the found between the VSP and Scott Henry (see Appendix 1). 2001/2002 low vigour vines produced wines for either of the attributes tested. higher concentrations of skin anthocyanin For the low vigour treatment, overall Dr Sally Bell participated in three Research to than high vigour vines, regardless of trellis astringency was higher from the Scott PracticeTM Grapevine Nutrition workshops at type. Low vigour Scott Henry (SH) vines Henry wine, but no difference in fruit Langhorne Creek (SA), Mildura (Vic) and also produced higher seed catechin and aroma was found. While the grapes from Naracoorte (SA) and in two Research to epicatechin concentrations than those from the two low vigour treatments were picked PracticeTM Winegrape Quality Management high vigour SH vines. The smaller berry at similar ripeness levels, the wines from workshops at Great Southern (WA) and Swan weight from the low vigour vines was the Scott Henry trellis treatment were Valley (WA) (see Appendix 1). She further a contributing factor to the higher higher in alcohol content than the VSP prepared two articles entitled Late season concentrations observed. However, vigour wines by approximately 0.7%, and the chemical control options for Botrytis and Why had little effect on seed catechin and ripeness difference may account for the are recommended export withholding periods epicatechin concentrations in Vertical Shoot astringency difference observed. different to the withholding period stated on Positioned (VSP) vines. The difference in berry the product label? for Technical Review and weight between high and low vigour VSP These results have to be confirmed with the September 2002 Murray Valley Winegrape fruit was less than that of the high and low data currently being acquired from the Growers newsletter, respectively. vigour SH fruit which may account for the 2002/2003 season. lack of effect of vigour on VSP seed In the 2000/2001 season, a study procyanidin concentrations. commenced which aimed to investigate the effect of plant water status and canopy report Teams’ Services Industry 47 48 Industry Services Teams’ report chapter onwine and healthwasalso the riskofall-causemortality. Abook moderate consumptionofwinereduces epidemiological evidencethatthe low to biological mechanismssupporting the ‘French Paradox’in1992onthepotential generated sincethepublishingof the the extensiveanimalandhumandata New York. This bookchapterreviews published byM/SHaworthPress Inc., edited byDrV.K. Joshi,whichistobe principles, practicesandrecentinnovations book entitled perspective wine fromaclinicalandscientific entitled Manager coordinated. Abookchapter Health andRegulatoryInformation interaction withvitaminE b components cardioprotective activitiesofwine Potential GWRDC-funded project HRI97/2 from the (accepted forpublication);thispaperensues the apolipoprotein Egenedeficientmice red winedecreaseslesiondevelopmentin Institute, apaperentitled O’Halloran, formerlyoftheHeartResearch Professor RolandStockerandDrRuth alsoco-authored with Information Manager & Winemaker the Education, inadditiontotwopapersfor newsletter oftheAustralianSocietyWine moderation publication, international have beenprepared forthebimonthly and printmedia.Forexample,three articles have alsobeenprepared fortheelectronic newsletters. Articlesandothermaterial alcoholindustry industry andinternational Review in theInstitute’s publication, and articleshavebeenprepared forinclusion health effects ofwinehasbeenadded to scanned, thedatabaseofresearch onthe havebeenregularlycontinued. Thejournals relevant has medicalandotherjournals and thegeneralpublic.Subscriptionto and healthissuesfrom industry, government Regulatory InformationManageronwine requests were received bytheHealthand During theyear, 96independentinformation both nationalandinternational. between complementaryorganisations, formal andinformalexchangeofinformation andbythe medical andscientificjournals, facilitated bythesubscriptiontorelevant MemorialLibrary.Fornachon Thiswas databaseoftheJohn on theinternal effects ofalcoholandinparticular, wine, the beneficialanddetrimentalhealth responsibilities, adatabaseofresearch on 1991, andhasestablished,aspartofher Regulatory InformationManagersince has assumedthepositionofHealthand Ms Creina Stockley, aclinicalpharmacologist, Staff: health issues Preparation ofinformationonwineand Australian &NewZealandGrapegrower American JournalofClinicalNutrition Creina Stockley, PeterHøj , andforotherAustralianwine The potentialtherapeuticvalueof , andonearticleforthequarterly has alsobeenprepared forthe ased onsynergistic . The Health andRegulatory The . Handbook ofenology: , whichthe AIM—Alcohol in De-alcoholized Technical for , outcome of the GWRDC-funded research is of outcome and selectfuture projects. Theprimary well asalistofcriteriawithwhichto evaluate research prioritiesforwineandhealth, as projects based onaprepared listofAustralian report, whichalsoincludedpotentialnew outcomes havebeendocumented inafinal workshop presentations andresearch the research/scientific community. The participants represented bothindustryand organised byCreina Stockley, andinvited The workshopwasdesigned,developedand further direction forresearch inthisarea. outcomes forwineandhealthtoprovide to summarisetheGWRDC-fundedresearch A workshopwasfundedbytheGWRDC Workshop http://www.gwrdc.com.au. reports canbeviewedon have nowbeencompletedandthefinal interaction withvitaminE components basedonsynergistic cardioprotective activitiesofwine consumption ofwine free radicalsbytheregularandmoderate damage toLDLandDNAfromoxidative peroxidation inhumans,Reductionof phenolics: absorptionandinhibitionoflipid consumption entitled projects onmedicalaspectsofwine role inthree GWRDC-fundedresearch played acoordinating andaparticipating Through Creina Stockley, theInstitute has Project coordination and inthe publicised inthe also beenfavourablyreviewed and to LallemandinFrance.Thebooklethas health andalliedprofessionals, and to thefoodandbeverageindustry, to door andothersalesoutletsinAustralia, distributed tothegeneralpublic,cellar approximately 48,500bookletshavebeen and printingcosts.Asof1July2003, contributed towards itspublication favourably reviewed thebookletand Department ofHealthandAgedCare and published.TheCommonwealth on wineandhealth A bookletentitled etduVin. de laVigne General AssemblyoftheOffice International which waspresented atthe83rd 2003 OIV NutritionandHealthSubcommission, proposal onapreferred structure forthe Nutrition andHealthSubcommission,a consumption andyoungadults OENO/ASP/01/190/Step 5 00/150/Step 6 wine andscientificresearch OENO/VINSAN/01/191/Step 7 OENO/ASP/99/129/Step 7 wine industryincludedraftresolutions Submissions prepared fortheAustralian Peter Gago. wine stylesandtastes prepared forthebookentitled Cairnes Post. OIV/WHO relations Sydney MorningHerald The A-Zofinformation was developed , and Grape antioxidant by PatrickIlandand . Allthree projects Wine Health warning Potential , OENO/SCVNS/ Specificity of Australian for the and , Prime Minister’s Science, Innovationand Horticultural SocietyofSouthAustralia; Committee oftheRoyalAgricultural and Australian Premier’s Committee;Wine Wine PublishingInc.;South for Viticultural Provisor PtyLtd;ManagementCommittee Centre forViticulture; Board memberof member oftheCooperativeResearch Advisory Committee(AWBC/WFA); Board Industry Technicalboards duringtheyear:Wine served onthefollowingcommitteesand In furthersupportoftheindustry, theDirector Høj istheChairof12thAWITC. Viticulture andOenology. Professor Peter conjunction withtheAustralianSocietyof Industry Technical Conference (AWITC), in facilitating thetriennialAustralianWine Australian wineindustryisitspivotalrole in aspects oftheInstitute’s supportofthe Advisory Committee.Oneoftheimportant the AWBC/WFA IndustryTechnical Wine Trade AdvisoryCommittee(onrequest) and membership ontheAWBC International industry hasbeenderivedfrom theDirector’s During theyear, additionalsupporttothe Industry Committeemembership of bodiesandcompaniesonanongoingbasis. contributes adviceoftechnicalnature toahost elsewhere inthisReport).TheDirector of theIndustryServicesteamare reported Regulatory InformationManager(activities and industrywere fieldedbytheHealth and and regulatory issuesfrom thegovernment information requests onscientific/technical until 30June2003,206independent team andtheDirector. From 1July2002 Information Manager, theIndustryServices industry, through theHealthandRegulatory advice andassistancetotheAustralianwine to provide legal/regulatory andtechnical One oftheactivitiesInstitutehasbeen Information requests Staff: Australian wineindustry Technical andregulatorysupporttothe recommended byWorkshop participants. those withtype2diabeteswasalso cardiovascular disease-theobeseand hi population groups at cardioprotective intwo is consumption determine whethermoderatewine confirm theseresults. Clinicalresearch to recommended furtherclinicalresearch to antioxidant, vitaminE.Workshop participants synergistically withtheendogenous act phenolic compounds,whichmay antioxidant effects ofthewine-derived the influence to status, however, appear Background diet,disease may bedealcoholisedred wine. atherosclerosis andcardiovascular disease, density lipoprotein (LDL), theprevent and reduce oxidation of low to efficacious beverages tested,themost are absorbedintothebodyand,of compounds phenolic that derived the wine- Peter Høj,Creina Stockley gher riskof associated with and smoking export markets provisions forwineinAustraliaand its Federation ofAustralia.Aguide conjunction withtheWinemakers’ Food Standards AustraliaNewZealandin and NewZealand 2 oftheFoodStandardsCodeAustralia amendments toStandard4.1.1of Volume Food StandardsCodeandconsequential change StandardP4ofthe[Australian] further amendmentsentitled Code ofAustraliaandNewZealand, provisions forwineintheFoodStandards towards thelabellingandproduction and resources havecontinuedtobedirected During 2002/03,significantefforts, energies Reviews andpublications industry’s operatingenvironment. societal issuesrelating tothewine commercial, environmental, politicaland exposes studentstoorganizational, students enrolled intheCourse,which Adelaide (Appendix2).Initsninthyear, 62 atTheUniversityof Agriculture andWine and communication 3005WT Information Manageralsocoordinate Course The Director andHealthRegulatory NutritionanditsHealthSubcommission. Vin du et delegate forOffice delaVigne International Environment Committee);andwasAustralian Working Group (Sub-committeeofSAWBIA’s Environment Committee;theEco-efficiency the AWBC/WFA IndustryNational Wine the AWBC LegislationReviewCommittee; Advisory Committee(asTechnical Liaison); IndustryTechnicalcommittees: Wine Manager wasamemberofthefollowing The HealthandRegulatoryInformation Premier’s ScienceandResearch Council. Engineering Council;SouthAustralian Grape industry practice,policy Grape industry , hasbeendrafted andwill have beensubmittedto for theSchoolof Application to Winemaking was published(Institutepublication #715). process to theconcentrationofleadinwine paper onthecontributionofwinemaking meeting inFranceJune2003.Another Pretorius presented thesepapersatthe OIV meet theconsumerchallenge current statusandfutureopportunities to . B P. Pretorius, I.S.;deBarros Lopes,M.A.;Høj, in thedevelopmentprocessofwine use ofnon-modifiedGMO-derivedproducts Høj, P.B.; Stockley, C.S.;Pretorius, I.S. etduVin: Office delaVigne International to the83rd 2003GeneralAssemblyofthe and industry, twopaperswere contributed winemaking. Attherequest ofGovernment potential ofprocessing aidsusedin of bromide ionsinwine;andtheallergenic ethyl carbamateingrapesandwine;sources barrels andstaves;regulations relating to the useofdiscrete piecesofoakversus toxicological andtechnicalissuesrelated to natural versussynthetictartaricacid; soot-contaminated grapeswithethanol; consequences ofsprayingandwashing an agrochemical; regulatory andscientific regulatory consequencesofusingmilkas oil impregnated waxinwinemaking; and technicalconsequencesofusingmustard Producers’World Group; Wine regulatory Forestry Australia(AFFA) delegationto limits fortheAgriculture, Fisheriesand Australian winemakingandtheirmaximum additives andprocessing aidsusedin reviewed, andmaterialprepared includes: information and/orissuesthathavebeen During 2002/2003,scientificandtechnical 1995 and2001. Australian wine Analytical specificationsforexportof complements theInstitute'sotherguide early inthenextfinancialyear;this be availableontheInstitute’s websitevery Development ofsuperiorwineyeast Sally Bell & Creina Stockley Sally Bell&Creina , whichwaspublishedin . Professor ; and The : bimonthly publication, all grapeandlevypayers,bytheInstitute’s to theAustralianwineindustry, including Technical Informationisalsodisseminated allergenic proteins inthewineswillbe antibodies. Furtherevidenceofdetectable commercial polyclonalandmonoclonal already beendevelopedforovalbuminusing allergenic proteins inthewines;anELISAhas (ELISAs) todetectwhetherthere are development ofenzymeimmunoassays isinglass) isbeinggeneratedforthe (ovomucoid, casein,ß-lactoglobulin,and a panelofrelevant monoclonalantibodies of processing aidsinthewines.Forexample, identify andquantitatetheconcentration assays are currently beingdevelopedto been collected,andsensitivespecific hundred commercially availablewineshave from therelevant ethicscommittees.One human ethicsapproval hasbeenreceived to eggs,fish,milkandnuts.Animal ingestion insubjectswithconfirmedallergy section isacontrolled laboratorytrialofwine allergenic proteins inwine,andthe second section isthedeterminationofdetectable The project comprisestwosections. Thefirst the labelofeachwineandproduct. winemaking requires declarationofuseon isinglass andmilkproducts in use oftheprocessing aidscasein,egg white, Standards AustraliaNewZealand,where introduced on14December2002by Food requirements forpotentialallergens was developedinresponse tolabelling is theprincipalinvestigator. Theproject supervisor andProfessor RobynO’Hehir Hospital andCreina Stockleyisthe project funded bytheGWRDCthrough The Alfred commenced on1July2002.Theproject is Research Institute The AustralianWine Immunology, TheAlfred Hospitaland Department ofAllergy, AsthmaandClinical A collaborativeproject betweenthe allergens inwine The identificationandmeasurementofpotential several years. Furthermore, theseissuesoftenspan both inAustraliaandinternationally. regularly raisedbyindustryorgovernment, ongoing astechnicalandregulatory issuesare support totheAustralianwineindustryis This project oftechnicalandregulatory staff (Appendix1). the manypresentations givenbyInstitute regulatory andtechnicalissues;through the Institute’s EmailAdviceBulletinon the Institute’s website(www.awri.com.au); Australian grapegrowers andwinemakers); MemorialLibrary(availableonlyto Fornachon web-accessible databaseoftheJohn regions (Appendix1);through theremote Institute’s Roadshowsinvariouswinemaking Communication andPublicityManager;the Report This isinadditiontotheInstitute’s (see furtherdetailselsewhere inthisReport). Manager waseditoruntil1January2003 which theHealthandRegulatoryInformation edited bytheDirector andthe Technical Review Annual , of 49

Industry Services Teams’ report 50 Industry Services Teams’ report Analytical Service, overseeingmanagement qualitymanagementsystemsofthe internal Manager, takesprimaryresponsibility forthe Mark Gishen,theInstitute’s QualityLiaison management standard. full ISO 9000 quality and leadingtothe Codex HACCPprinciples the from starting recognisedof internationally standards, the in uses astagedapproach cheap program that relatively and simple From grapestoglass having attended(nowtotalling94).The another sixtothegrowing listofcompanies was conductedthroughout theyear, adding Only onecoursefortheHACCPmodule smaller scalebusinessesintheindustry. regulations, andmeettheneedsof (now delayed)changestothefoodhygiene satisfy therequirements oftheproposed safety plan.Thismodulewasdesignedto analysis andcriticalcontrol point)food incorporates anHACCP-type(hazard delivered inaone-daycoursethat HACCP module-asimpleprogram Industry interest remains greatest inthe with asimpleHACCPmodulein1999. published inAugust1997andenhanced grapes toglass techniques toindustryremains the the provision ofadviceonqualitymanagement The majoroutputoftheInstitute’s activitiesin reported elsewhere intheAnnualReport. team leader. Thedetailsofthisproject are primary responsibility fortheproject as wine andgrapespirit,continuestotake compositional parametersingrapes,must, the rapiddeterminationofanumber use ofnearinfrared spectroscopy (NIRS)for collaborative research project evaluatingthe Mark Gishenremains heavilyinvolved inthe Quality LiaisonManager labelling forwine. articles intheprintmediaonallergen conjunction withthepublishingoftwo Professor RobynO’Hehir(Table 13),in been conductedbyCreina Stockleyand television andfiveradiointerviewshave (Institute publication#705),andone Zealand Grapegrower&Winemaker and publishedin background oftheproject hasbeenprepared One manuscriptoutliningthescopeand project willbecompletedby30June2004. the studyallergens.Itisanticipatedthat and intheabsenceofspecificIgEtoany without ahistoryoffoodorwinereactions compared toagroup of10healthysubjects and clinicallycharacterised,theywillbe 30 challengesubjectshavebeenrecruited section oftheproject. Twenty-one ofthe with theclinicalchallengesofsecond wines usingBAT isoccurringintandem and theprocessing aids.Analysisofthe100 developed andoptimisedforusewithwine to provide abloodsample.BAT havebeen relevant foodallergensare beingrecruited subjects withextreme sensitivitytothe (BAT) andflowcytometry, andchallenge obtained from basophilactivationtests program, whichwas The Australian&New program provides a attainment From Library canbefoundelsewhere inthisReport. Library. Thereport oftheactivities Memorial output oftheJohnFornachon also responsible fortheperformanceand Communication andPublicityManager is and toactasaconduitformedia. The and othernon-technicalcommunications, coordinate theInstitute’s printed material plan objectives.Partofherrole isto Institute’s missionstatementandbusiness the with line positioning isdevelopedin positioning (value)ofthe Institute. This groups receive aclearunderstandingofthe for ensuringthatindustryandstakeholder and PublicityManager, sheis responsible Technical Information) be foundunderthereport for Conference (detailsofherotheractivitiescan IndustryTechnicalthe AustralianWine Director andtheConference Managerof who isalsothePersonalAssistantto role isapart-time positionforRaeBlair, The CommunicationandPublicityManager’s Communication andPublicityManager aims. its has draftedaworkprogram toprogress and prioritised thevariousareas tobeaddressed the paymentforwinegrapes.Thegroup has for wineries by used measurement instruments develop a metrologicalfor system control CommissionStandards National and aims to formed inresponse toaproposal putbythe gr This as theLegalMetrology Group. group known (WGCA) recently formed working Growers' CouncilofAuand Winegrape (WFA) FederationofAustralia the Winemakers' on Mark GishenistheInstitute'srepresentative management systemsimplementation. as partofageneralmovetoward quality of policyandprocedural documentation, as ameansofassistingcontrol andavailability throughout theInstituteingeneral, primarily The software isgraduallybeingintroduced the quality managementsystemwiththeaidof Analytical Servicecontinuestomanageits and foragrochemical residue testing.The testing programs forroutine wineanalysis proficiencynational andinternational continues toparticipateandexcelinboth for agrochemicals. TheAnalyticalService FARA, andtheHPLCmulti-residue method wine byenzymatickitusingthe being thedeterminationofaceticacidin expanded toincludetwomore methods In addition,accreditation ofmethodswas extended tocoverhydrometer calibrations. Matt Holdstock'ssignatorystatusbeing of thechemicaltestingprocedures, and of AmandaCookasasignatoryformost time ofreassessment includedtheaddition Several changesthatwere madeatthe was grantedcontinuingaccreditation. during December2002andthelaboratory NATA againsttheISO17025standard of theAnalyticalServicewasreassessed by of itsNATA accreditation. Thequalitysystem role ofAuthorisedRepresentative inrespect corrective actions,andhasnowtakenonthe reviews, documentation,auditing,and Paradigm Quality software package. . AsCommunication rvso of Provision Cobas oup was stralia’s > > enquiries received. Manager anddonotrepresent thetotal responded tobytheCommunication represent thoseenquiriesthatwere without forwarding. Thesefigures only there are manythatcanbedealtwith forwarded totherelevant expert,however, Many requests forinformationare simply as the‘generalenquiries’contactaddress). information (asheremailaddress is listed 51 Manager receives regular requests for widely knownandused,theCommunication 19 theInstitute’s*With websitebeingmore 2 Total 1 7 Other 1 Media 11 Government Consultants Industry Students during theyear: Communication andPublicityManager General enquiries*responded tobythe and distributed: The followingpress releases were prepared media overtheyear(Table 13). representatives from variousformsof Institute staff were interviewedby Media contactsandinterviews Appointment ofPeterWall asChairman, Appointment oftheDirector ofResearch, Officer ofProvisor. and DrDarren OemckeasChiefExecutive Professor SakkiePretorius Table 13.MediainterviewsofInstitutestaff for2002/2003 010 ee ø uuerl fRDfrwn nutysces>Jeremy Oliver >Tyson Stelzer, www.cellaringwine.com >AlexiaMoses, Future role ofR&Dforwineindustrysuccess Qualityfactorsforsparkingwine/Champagne PeterHøj Ageing ofred wineunder screw cap 20/1/03 PeterGodden 30/12/02 PeterGodden 30/12/02 details Media Itemdiscussed Staffmember Date 590 ee odnCr n C,wt atclrrfrnet >UK freelance journalist, Jamie G CorkandTCA,withparticularreference to PeterGodden 25/9/02 9/12/02 Peter Godden Screw caps and the closure trial > Bernard Carpinter, >Bernard >JoBurzynska,CrierMediaGroup, UK, Closure permeation–reference towine Screw capsandtheclosure trial PeterGodden PeterGodden 23/12/02 9/12/02 81/2SlyBl rp rp n aiiy>GuyWebber, >GrahamO'Neil,Australian >MaxAllen, Grape crops andsalinity Attemptstodevelopvariousmethods PeterGodden Biotechnologyinwinemaking 2/12/02 SallyBell 28/11/02 Miguelde 28/11/02 PeterGodden 15/11/02 of >RobertMcClean,arrangedbyBelindaByrne 26/9/02 GWRDC-fundedallergenstudyandwine 25/9/02 Creina Stockley 24/9/02 790 riaSoke elhbnft fse/knerce ie>DanNolan, >Jeremy Roberts, Discussionofresearch findings(USA)thatglass Healthbenefitsofseed/skinenrichedwine PeterGodden Creina Stockley 31/10/02 27/9/02 >NickiBourlioufas, Informationforarticleonclosures and PeterGodden 19/9/02 13/9/02 970 ee odnCouetil>Emailed:MichaelJames,ABCNEWS.com,USA >MichaelSexton, >MaxAllen, >Instudiorecorded interviewwithAlanSaunders, >SusieBarrie, Closure trial JamieGoode, >UKfreelance journalist, PeterGodden 30/7/02 Changesthatoccurinwinewhendecanted 29/7/02 NIRproject Corkissueswithemphasisonclosure trial PeterGodden GMOsandthewineindustry MarkGishen 17/7/02 PeterGodden 15/7/02 36monthresults from theclosure trial PeterGodden 12/7/02 11/7/02 PeterGodden 8/7/02 /10 iuld ars eei oiiain>GrahamLawton, > Provided informationtoLynn Alley, Spectator Wine >AlexandraRundel,Editorfor Geneticmodification Screw capusageinAustraliaandNZclosure MigueldeBarros >BirgittBaader, 8/11/02 ReJamieGoodes’articlein PeterGodden >JessicaLawrence, 8/11/02 Allergenlabelling Discussionof36monthresults from closure trial PeterGodden Creina Stockley 4/11/02 PeterGodden Allergenwinelabelling –originsofandreasons1/11/02 1/11/02 PeterGodden 31/10/02 290 ee odnAR/odhw>DavidKing, >SpokewithMaxAllen Television > Win (Channel9)on6pmNewsthroughout BruceMounster, > >BenCanaider( SallyEaston, >UKfreelance journalist, AWRI/Roadshow Allergensinwine PeterGodden 12/9/02 Furtherinformationonarticlechanges that PeterHøjand 5/9/02 PeterGodden SallyEaston, >UKfreelance journalist, 4/9/02 >JoBurzynska,CrierMediaGroup UK,Editorof inthewineindustry GMOs AWRI/Roadshows AWRI/Roadshows >BruceMounster, PeterGodden >RobynSmith,ABCTVNewsandcurrent affairs PeterHøj 29/8/02 Discussionofthe36monthresults from PeterGodden 26/8/02 Closures andbottleageingofwine 23/8/02 Issuesrelating to PeterGodden PeterGodden 23/8/03 Therole oftheAWRI, withreference to PeterGodden 21/8/02 Resultsofclosure trialandclosure issues 19/8/02 PeterGodden PeterGodden 15/8/02 14/8/02 8/8/02 Peter Godden Incidence of TCA in wine sold in Australia with > Tim White, >Tim IncidenceofTCAinwinesoldAustraliawith PeterGodden 8/8/02 Barros Lopes Lopes Creina Stockley eeomn n rdcie rmsEditorof development and‘reductive’ aromas for theremoval ofTCAfrom corks Brettanomyces labels. industry positiononallergenwarning discussion ofclosure trial KWn n prtAscainsuypublishedin andSpiritAssociationstudy UK Wine n eae,addfeecsi esr hostof perception attributabletotheshapeofglass and aerated,differences insensory attributed toPWG. for usingcertainwineadditivesandprocessing aids susi eea (NewYork, USA) issues ingeneral shape hasaneffect onwineflavour occur inbottledwine the closure trial odhw,adtecouetil(published23August2002) Roadshows, andtheclosure trial Assessment Course reference tostatisticsfrom the and wine > Tim Atkins, >Tim and wine Brettanomyces Harpers Advanced Wine Advanced Wine n ie>PhilipRich, and wine n uts >DavidBird, from Freelance UK journalist and quotes > DavidAnderson,ABCRadioQueensland AustraliaandPerth > LiamBartlett,ABCRadioWestern > This isinrelation tothetwoarticlesin > > SusieHamilton,ABCRadioRenmark > DerrynHinch,3AK(Melbourne) > CeinwenParish,RadioNewZealand > > ABCRadioDrive,broadcast on29thAugust > Live-to-airinterviewwithKevinNorton,ABCRadio, Australian The BendigoTimes Channel 9News magazine, December2002 Island newspaper Wineanorak.com, Wineanorak.com, and ABCCapricornia Drinks Business Friday 30thAugust throughout exceptcentral Melbourne Victoria andGeelong exceptgreaterVictoria Melbourne Drinks BuyerEurope (published 10and17August) Adelaide The ComfortZone Drinks BuyerEurope Drive (14 and15/9/02) Weekend Australian The Weekend Australian Australian FinancialReview Australian FinancialReview Channel 10News Guardian/Sunday Observer The Press Decanter Harpers The StockJournal Sydney MorningHerald Choice Age Epicure magazine The Australian 7:30 Report Tasmanian Country Tasmanian Country (screened 25/9/02) New Scientist, Sunday Mail Liquor Watch Harpers , storywastoberunon The DominionPost, and referred toin - theChristchurch/South ) ABC radio(aired 20/7/02) Biotechnology News Saveur (ABC TV) , Brisbane (Brisbane) The Australian oode, Wineanorak.com, magazine, UK United Kingdom magazine The Weekend newspaper newspaper Drinks Business , UK Wellington NZ Decanter (25/9/02). 51 Industry Services Teams’ report 52 Industry Services Teams’ report 13/1/03 Peter Godden Effect of increased alcohol on the perception of > Tim White, >Tim Effect ofincreased alcoholontheperception of PeterGodden 13/1/03 020 ee odnPossible effect of rain on the wine i >Andrew McGarry, Comment on Godden Peter 20/2/03 PeterGodden Effect ofdrought andveryhightemperatures on 7/2/03 PeterGodden 17/1/03 Date Staff member Item discussed Media details Media >JancisRobinson >JessicaLawrence, labelingwithregard Wine toallergens /origins >Tamara Hunter, Various issuesregarding ageandyieldof PeterGodden 3/1/03 PeterHøj Pastandfuture directions andoutcomesof Item discussed 6-8/2/03 Creina Stockley Staffmember 30/1/03 Date 820 lnPlnt a > White, >Tim >MarkChipperfield, InformationforarticleonPiedmontese, Oak PeterGodden AlanPollnitz 5/3/03 Closure issues,withparticularreference to 28/2/03 PeterGodden 27/2/03 //3PtrGde h s fsprrtclC2t eoeTA >GiselleWeiss, >KristiEssick, >Louise Marr, Vierra, >Tina ABC Theuseofsupercritical CO2toremove TCA >KirstyWilcox, Technologies usedintheproduction of PeterGodden 8/4/03 Bushfire smoketaintingrapes/wines PeterGodden 4/4/03 PeterGodden Percentage ofTCAtaintedwinesinAustralia 2/4/03 Informationonclosure trial 1/4/03 PeterGodden PeterGodden 26/3/03 20/3/03 930 ee odnGnrlpreto fvnae atclry >MaxAllen, >KatieFisher, Melbourne Weekly Times, Generalperception ofvintage,particularly PeterGodden 19/3/03 Effects ofbushfire smokeongrapesandwine PeterGodden 18/3/03 PeterGodden 17/3/03 7/3/03 //3PtrGde lsr ri,adcr/lsr sus>CyrilPenn, >JamieGoode, Closure trail,andcork/closure issues PeterGodden Methodsofremoving TCAfrom cork PeterGodden 9/5/03 PeterGodden 28/4/03 28/4/03 260 ee odnIse eaigt h fet fbsfr >KathyBolt, Issuesrelating totheeffects ofbushfire PeterGodden 12/6/03 //3PtrGde h s fpoenaayi o aitl >MaxAllen, Theuseofprotein analysisforvarietal PeterGodden 6/6/03 //3PtrGde ees soi ehdt eoe >KatieFisher, >Tejinder Gill, Reverseosmosismethodtoremove PeterGodden 5/6/03 Closure issues PeterGodden 20/5/03 26/5/03 Peter Godden Objective and subjective measurement/definition > Tim White, >Tim Objectiveandsubjectivemeasurement/definition PeterGodden 26/5/03 26/5/03 660 ee odnTeueo rti nlssfrvrea >MelodyHorrill, Theuseofprotein analysisforvarietal PeterGodden 16/6/03 860 dniiaino uc n ie>Tony Love, identificationofjuiceandwine Issuesrelating to PeterGodden 19/6/03 18/6/03 //3PeterGodden 4/6/03 //3PtrGde ore fTAi ieadsaitc n >JoyWalterfang, Sources ofTCAinwineandstatisticson PeterGodden 2/6/03 6/6/03 Peter Høj Australian wine industry > Willy Billiard, >Willy wineindustry Australian PeterHøj 6/6/03 and processing aids. 5 January2003. 5 Alsoarticlein and processing aids. of andreasons forusingcertainwineadditives acceptance ofscrew capsforred andwhitewines h oeta ult f20 avs included inanarticlepublishedon18January2003) the potentialqualityof2003harvest flavour andaroma inwine. Australian vineyards research intohealthbenefitsofred wine Australian winesoldintheUK. synthetic closure useinAustralia,andfor grapes andwine. bushfire smokeissue Brettanomyces atclryBrl,wns (Full-pagestorypublished8March). particularly Barolo, wines. Brettanomyces requirements ofwinebottleclosures ingeneral. from cork,andotherissuesregarding thetechnical identification ofjuiceandwine bsfr mk an’fo ie>Liz Rodway, ‘bushfire smoketaint’from wine and consumeracceptance Closure issues,particularlyscrew capindustry Debate abouttheuseofoakchipsversusbarrels. smoke ongrapesandwine. Brettanomyces incidence ofTCAtaintinwine. fwn qaiy.(Full-page storypublished31May) of wine‘quality’. in red wines wine, andsensorythresholds forfour-ethylphenol Choice issues, particularly control strategy > JamesKelly, > issues, particularlycontrol strategy Dekkera/Brettanomyces magazine survey on consumer > Kim Arlington, Associated Press Associated Arlington, Kim > magazine survey on consumer ndustry and vintage > Jessica Rich, Rich, Jessica > ndustry andvintage and > Tim White, > JamieGoode,articlefor > JennyPollack, > ChrisSnow, Herald > HuonHooke > KylieStevenson, Sydney MorningHerald (aired 20February2003) discussed on Age Grapegrower &Winemaker magazine andwebsite,USA New Zealand Wine Industry Journal Industry New ZealandWine (articles published1April2003).Articlealso and Melbourne Age The Advertiser Wine BusinessMonthly Wine Harpers Magazine The Weekend Australian Australian FinancialReview Australian FinancialReview Australian FinancialReview Australian FinancialReview Australian FinancialReview Practical Winery &Vineyard USA Practical Winery ABC Radio The Australian&NewZealand Drinks Bulletin Weekly Times The Wall StreetJournal Channel 9NetworkNews The Australianand Sydney MorningHerald ABC NewsRadio Vigne etVin Uncorked Magazine The Economist Sunday Mail Harpers Bent Channel 10News The West Australian Winestate Winestate Liquorwatch Radio 666Canberra Sunday Mail The Weekend Australian Sunday Telegraph magazine magazine, UK. , SheppartonVictoria Harpers ) , Adelaide International Publications International , Adelaide, newspaper, Melbourne magazine magazine, UK , European bureau , Brisbane. on 1April2003. magazine , Adelaide , ( Sydney Morning , livetoairinterview , European bureau , London and Melbourne (Information Analytical Service

Staff: Don Buick, Matthew Holdstock, Greg Ruediger, Amanda Cook, Matthew Cream, Randell Taylor, Sandra Lloyd-Davies, David Boehm, Athina Massis, Anna Catalano, Danielle Leedham, Maria Mills, Heather Brooks and Belinda Bramley. Peter Eichinger commenced in June 2003.

The Analytical Service (AS) is divided into three sections namely the analytical laboratory supervised by Matt Holdstock, the trace analysis laboratory supervised by Greg Ruediger and the administration section supervised by Sandra Lloyd-Davies.

The AS provides a commercial testing facility operating independently within the Institute. It is self-funding in all respects from its fee earning work, and any surplus generated is returned to the Institute for the benefit of the Australian wine industry. It is focussed solely on providing services to the Australian wine industry, with a commitment to providing many specialised services that are not generally available in L to R Back row: Athina Massis, Maria Mills, David Boehm, Randell Taylor, Matthew Cream, Matthew Holdstock, Anna Catalano winery laboratories, or from other commercial Middle row: Greg Ruediger, Sandra Lloyd-Davies, Amanda Cook, Heather Brooks, Danielle Leedham, Jelena Jovanovic Front row: Peter Eichinger, Belinda Bramley laboratories. AS interacts with, and provides analyses for other groups within the Institute the previous year. Significant increases Sandra Lloyd-Davies, has proven to be very (Industry Services and Research groups) on a occurred in the number of AWBC certificates capable and adaptable. In light of these fee for service basis, although this constitutes and EU certificates, but the number of departures and staff development, the a minor part of the AS activities. Japanese certificates has declined due to signatory status of Matthew Cream has sluggish economic conditions there. It been upgraded and Peter Eichinger has Our aim is to provide a range of quality should be noted that some of the major attained accreditation for all tests. analytical services that are important to exporters have developed NATA accredited industry which can be provided in a reliable, laboratories and are thus able to issue their Matt Holdstock, Analytical Service Supervisor, price competitive, accurate and timely manner. own analytical certificates. Testing for has completed his studies towards a During December 2002, a market research chloroanisoles and 4-ethylphenol increased Graduate Diploma in Oenology. The Service, survey was commissioned (McGregor Tan with the latter being related to growing therefore, now has two qualified oenologists Research) in which about 200 wineries and concern with Brettanomyces spoilage of to help enhance the service and advice that suppliers to the industry were questioned wine and the associated Industry Services can be offered to industry. Matt’s completion on their knowledge of the AS’s services and investigation. The number of data points of his studies will allow him to provide performance. The results of the survey produced increased by 29% indicating greater depth in dealing with client enquiries were very positive, with a very high (96%) many of the increased testing numbers related to the services we provide. Matt has awareness of the laboratory’s existence and were for multiple analyte testing. The also been involved with the Inter-Winery the range of services offered. The clients identified their key requirements being Table 14: Comparison of selected tests performed by period accuracy, reliability, strong technical expertise, NATA accreditation and timeliness, as well 98/99 99/00 00/01 01/02 02/03 as good customer service, confidentiality and reputation. The AS was rated very highly Total number of tests 39,087 44,846 46,037 48,314 53,932 on all of these aspects of its performance. A new fee schedule was prepared and mailed out to levy payers early in the financial year. performance of the laboratory in a range of Analysis Group (IWAG). The purpose of Overall, the Analytical Service conducted analytical classes is given in Table 14. this group is to expedite inter-laboratory approximately 54,000 individual tests on proficiency trials and enable a comparison wine and grapes in 2002/2003. These There have been some significant staffing of results obtained in different laboratories services included export certification and changes in the last year. Most notable using a range of techniques. support for the quality control activities of has been the retirement of Don Buick, winemaking and viticulture. The growing the Manager of Analytical Services. He Improvements in quality systems are seen interest in grape quality has led to increasing has overseen considerable growth in the as being very important. To aid in tracking numbers of grape samples being tested for business, provided sound engaging document versions, we use a software colour, glycosyl-glucose, brix, pH and titratable leadership and overseen a number of program known as Paradigm II (further acidity over the last two vintages. While reforms in the laboratory to improve details are provided under the Quality many large wineries do have testing facilities, efficiency and quality. Dr Peter Eichinger Liaison Manager’s report). A number of we receive samples from growers and has taken over this role, having experience other functions are proving valuable, as the consultants to independently test grapes as in analytical chemistry and chemical research alerting facility has been set up to automate this often impacts on grower payments. and will be endeavouring to continue this tasks such as scheduling ongoing calibrations focus. After long and dedicated service, of instruments, method reviews, audits, etc. The number of tests performed by the John Hughes left Analytical Services in Analytical Service increased by 12% from September 2002, and his replacement, Analytical Service report Service Analytical 53 54 Analytical Service report on thisinstrumenttoo. she willgraduallybeabletorunsequences Agilent GC/MSplatformandsowe expect aspects ofthissoftware iscommontothe operating software (Chemstation).Many training coursesontheAgilentHPLC has developedquicklyandattended and hassettledinquickly. Heather Brooks laboratory toassistwiththeextraworkload Bramley wasrecruited asaTechnican inthat ably handledbyRandellTaylor. Belinda the traceanalyticallaboratory(TAL) wasvery the Aprilquarter. Hisrole asSupervisorof taken mostofhisLongServiceLeaveduring As previously mentioned,Greg Ruedigerhas saving device. has proven tobeagreat timeandlabour was installedinSeptember2002,andthis A Mieledishwasher(forlaboratoryglassware) users. from Winescan ongoing calibrationandotherexperiences meeting inNewZealandanddiscussedthe covered. MattHoldstockattendeda FOSS in thisarea toensure thesewineswillbe needed. We wishtoexpandthecalibrations represented inthecalibrations,andare curves. Sweetwhitewinesare poorly and red winesrepresented inthecalibration very well,withalargenumberofdrywhite for Australianwinesamplesare proceeding optimal forAustralianwines.Calibrations for European wines,theseare unlikelytobe yet tobeconfirmed.Whilecalibrationsexist The fullcapabilitiesofthisinstrumenthave 2000wasinstalledlate2002. Winescan theassistanceofFOSSPacific,a With maximum capability. is wellmaintainedandusedtoits training coursestoensure newequipment staff haveattendedspecialisedinstrument existing servicesinthenewyear. Several headspace capabilitieswillcomplement capabilities. Theadditionoftheautomated some methoddevelopmentutilisingtheSPME envisaged thatwewillbeabletoundertake backlog ofwork.Oncethisoccurs,itis reduce turn-round timesandthecurrent capabilities willbeextremely usefulto financial year. Intheshortterm,GC/MS which arrivedinthethird quarterofthe this timewithheadspaceSPMEcapabilities, This yearASpurchased asecondGC/MS, upon theInstitute. therefore hasnotimpacted and delayed, GLPOECD testing c for registration ofagrochemicals under Residue Association Leave takenbyGreg Ruediger. TheNational been delayedinpartduetoLongService before anaccreditation visitoccurs.Thishas under theseguidelinesisstilltobeassessed achieve compliance,andanauditedtrial time, significantprogress hasbeenmadeto occurred inSeptember2002.Sincethat agrochemicals usedinthewineindustry OECD GLPguidelinestoresidues testingfor information aboutimplementationofthe An advisoryvisitfrom NATA toprovide onditions hasbeen 2 (NRA) requirements (APVMA), whichisduetobegazettedshortly. Australian PesticideandVeterinary MedicineAuthority Wine showinSeptember2002. Wine a trophy forthebestRieslingat Adelaide quality ofAustralianwine,theASsponsored In continuingitssupportofimproving the research component. corks andscrew caps,aswellasmall involve client-submittedclosures, reference commencement. Thecommercial trialwill work withintwotothree daysof with staff workinginshiftstocomplete the (see fullreport under of varioustreatments toreduce smoketaint smoke damage,aswelltheeffectiveness methylguaiacol asindicatorsoftheextent (over 200)were testedforguaiacoland4- profound affect. Alargenumberofsamples growers wasaverted,smokeexposure hada WhiledirectVictoria. fire damagetothe grapes from thebushfires inNorthEastern Institute’s response tothesmoke-affected earlier inthisreport, anexampleofthisisthe members ofIndustryServices.Asoutlined frequently interactstrongly withthestaff The staff intheAnalyticalServicegroup financial year. continue forseveralmonthsintothenew organoleptic qualities.Thisworkwill these compoundsonfermentationand winemaking trialstoevaluatetheimpactof registration, aswellsmall-scale involved bothtestingofresidue levelsfor involving theuseofagrochemicals. Thishas involved inanumberofresidue trials The traceanalysislaboratoryhasbeen sulfur dioxideandOD analysis oflargenumbersfree and total The trial. closure testing ofacommercial winemaking techniques andofprocessingaidsfor (see report under research closure trialatthe48monthstage Services teamtoanalysesamplesfrom a Staff haveworkedcloselywiththeIndustry resulting frombushfires in grapesandwine,causedbysmoke into thenatureandameliorationoftaints 2 The NRAhasrecently undergoneanamechangetothe ), as well as initial and six month six and initial ), as well as Evaluation ofnewanalytical Investigations conducted 420 ). were performed FOR THE YEAR ENDED30JUNE2003 STATEMENT OFFINANCIALPERFORMANCE Profit fromoperatingactivities Expenses fromoperatingactivities Revenue fromoperatingactivities oa hne neut 0,3 291,229 608,239 291,229 608,239 Total changesinequity Profit fromordinaryactivities orwn otepne02,112 109,085 0 161,791 518,789 0 557,898 Borrowing costexpense 700,000 Travel expenses amortisation expenses Depreciation and 1,044,055 1,194,694 service expenses Administration andgeneral 0 281,279 to Provisor PtyLtd 3,876,256 340,644 assistance SA Government 4,281,915 700,000 operating expenses Analytical andproject Employee benefitexpense 1,224,875 1,507,854 474,403 Other revenue 580,282 Provisor PtyLtd 77,548 to Assistance bySAGovernment 96,170 Analytical Service Commercial research collaborations CRCV project funds Development Corporation Research and Grape andWine oo eils014,503 (5,323) 0 (12,720) Equipment Motor vehicles Net gain(loss)ondisposalofassets rjc ud ,2,1 3,849,613 4,324,916 Capital grants Project funds ulig00 0 364,200 438,420 Equipment Building 7,2 439,572 471,029 2,5 282,049 620,959 032002 2003 $$ AS AT 30JUNE2003 STATEMENT OFFINANCIALPOSITION TOTAL EQUITY TOTAL EQUITY NET ASSETS LIABILITIES TOTAL Total noncurrentliabilities Non currentliabilities Total currentliabilities Current liabilities A TOTAL Total noncurrentassets Non currentassets Total currentassets Current assets eevs9670966,750 966,750 4,051,727 4,659,966 43,298 Retained profits 67,411 Reserves 575,068 0 624,457 323,186 Provisions Provisions Provisor PtyLtd Unpaid investmentin 1,352,485 1,482,678 840,000 and repayable 840,000 Project fundsnotexpended Payables andaccruals 0 1,834,661 2,070,026 650,000 1,559,247 of Oenology 1,522,048 IndustryChair Australian Wine Investment InProvisor PtyLtd 64,729 300,761 Plant andequipment 68,745 1,192,000 511,693 Leasehold buildings 1,253,916 1,428,325 1,416,954 Other current assets Commercial bills Receivables Cash assets RV1,4 22,340 17,345 208,055 191,589 CRCV GWRDC SSETS ,2,1 5,018,477 5,626,716 5,018,477 5,626,716 2,201,246 2,706,666 2,157,948 2,639,255 7,219,723 8,333,382 4,233,908 5,082,074 2,985,815 3,251,308 032002 2003 74143,298 67,411 $$ 55 Abridged Financial report 56 Abridged Financial report FOR THE YEAR ENDED30JUNE2003 STATEMENT OFCASHFLOWS e ahpoie yodnr ciiis109241,459,453 1,059,274 Net cashprovided byordinary activities Changes inassetsandliabilities 291,229 608,239 920,798 1,428,325 Non cashflowsinoperatingprofit Profit from ordinary activities (120,000) 0 provided byordinaryactivitieswithprofit Reconciliation ofnetcash 507,527 37,091 Cash at30June (11,371) Cash at1July 300 Net increase (decrease) incashheld 0 Net cashprovidedbyfinancingactivities Repayment ofloans 0 (700,000) CASH FLOWSFROMFINANCING ACTIVITIES Net cashusedininvestingactivities (326,814) 0 and equipment Proceeds from saleofplant (347,017) (682,215) to Provisor PtyLtd Assistancepaid SA Government 700,000 (522,000) (61,916) Payments forplantandequipment Provisor Pty Ltd Payment forinvestmentin 57,727 Payments forcommercial bills 112,541 for Provisor PtyLtd Receipt from SAGovernment 6,196,141 7,665,762 CASH FLOWSFROMINVESTING ACTIVITIES (4,794,415) Net cashprovidedbyoperatingactivities (6,719,029) Payments tosuppliersandemployees Interest received Grants andotherincome CASH FLOWSFROMOPERATING ACTIVITIES (Increase) decrease inreceivables Increase (decrease) inpayablesand hre o(euto n)poiin 352135,906 439,572 73,502 471,029 Charges to(reduction in)provisions (Profit) lossonthesaleofplant Amortisation anddepreciation ud nxedd1872607,506 108,732 (5,580) (214,948) funds unexpended accruals andproject and prepayments n qimn 270(9,180) 12,720 and equipment 10065 (831,926) (1,070,645) ,1,5 1,428,325 1,416,954 1,459,453 1,059,274 032002 2003 $$ (120,000) 0 P.W. Polyphenols,‘pigmentedpolymers’andred winecolour:results from theTannin Godden M.J. Herderich and P.A. Henschke ..Dmeg plctoso ISi h ieidsr SR ln nuty 9Aug02 CSIROPlant Industry 15Aug02 2 McWilliam’s Maurice Roadshowseminar: Oxidationofbottledwine:therole ofascorbicacid,andbottlestorage J.M. Eglinton ApplicationsofNIRSinthewineindustry McWilliam’s MauriceO’SheaAward 2002acceptance speech E.J. Waters R.G. Dambergs P.B. Høj P.A. Henschke ..Dmeg ISi h ieidsr ir ir 27Aug02 Wirra Wirra Tannins andmouth-feel:terminology methodsforsensoryevaluation of red wines NIRSinthe wineindustry Theuse ofNIRforthemeasurement ofgrape, juiceandwinecomponents R.G. Dambergs Waters E.J. M. Gishen Eglinton) (presented byJ.M. P.A. Henschke of Microbiological 10-11Jul02 andeffect onwinecomposition G.H. Fleet ASVOViticulture E.J. Bartowsky, Date Ascertainingthedegree ofgeneticvariationbetweenstrains Whereandtowhom Barros Lopes Polyphenols,‘pigmentedpolymers’andred winecolour:results from the M.A. de M.J. Herderich Title oftalk P Staff talksandposterpaperspresentedbyInstitutestaffduring2002/03 Externalseminars, APPENDIX 1. ..HjFrom molecularresearch toenjoymentofwine– R.E. Day P.B. Høj SoiltypeandclimaticinfluencesonthewinesofBordeaux P.W. Godden ..FacsOxidation ofbottledwine:therole ofascorbicacid,andbottlestorage TheuseofNIRforthemeasurement ofgrape,juiceandwinecomponents P.W. Godden 25Jun02 (not Measuring desirableoakwoodcomponentsinwine Theinfluenceofdeficitirrigationonsaltaccumulationingrapes,andwinequality I.L. Francis OenologySymposium, Tannins andmouth-feel:terminologymethodsforsensoryevaluationofred wines M. Gishen A.P. Pollnitz Lallemandvisit, M. Gishen I.L. Francis P Yeast interactionwithgrapephenolicsandeffect onwinesensoryproperties and E.J.Bartowsky P.A. Henschke Soden, P.B. Høj Gockowiak, A. Francis, H. influenceonChardonnay winecompositionandflavour P.J. Costello,I.L. J.M. Eglinton, Costello, P.R. Grbin -.Bl Agrochemical issuesforgrapegrowers andwinemakers Qualitymanagementsystems–ISO9000,HACCPwhatare they, doyou need them? J.M. Eglinton Managingstuckfermentation S-J. Bell Gishen M. P.W. Godden P.A. Henschke UpdateontheAWRI trialofthetechnicalperformancewineclosures P Corktaint–causes,detectionandprevention FlavouraspectsofMLF–control ofthe‘buttery’diacetylcharacterinwine E.J. Bartowskyand P.W. Godden A.P. Pollnitz P.A. Henschke ..Brosy Microbial inducedoxidativespoilageofbottledred wine D. Xia E.J. Bartowsky, K.S. Howell J.M. Eglinton .A. Henschke .A. Henschke .A. Henschke 3 , G.H.Fleet 2 3 , Yatsri yaisdrn ie utr iefretto ItrainlUin27Jul02 Union International Yeast straindynamicsduringmixedculture winefermentation , and Winemaking with alternative yeasts:influenceonwinecompositionandflavour withalternative Winemaking and n Therole ofVA inrestarting stuckfermentation and , Winemaking withselectedstrainsofnon- Winemaking , .. assadcnrlo osns nwn odhwsmnrt:2 Jul02 Roadshowseminarto: Causesandcontrol ofmousinessinwine P.J. 3 , 1 h ie fSuensadBra Negociants Australia the needforanintegratedapproach andBarsac the winesofSauternes Geographic, viticulturalandwinemakingfactorswhichinfluence Winemaking with alternative yeasts:influenceonwinecompositionandflavour withalternative Winemaking The artandscienceofwinequality Dekkera/Brettanomyces Project, includingthe2001large-scalewinemakingtrial odtosnndsrciemaueeto ht ieclu Hobart,Tas conditions/non destructivemeasurement ofwhitewinecolour Dekkera/Brettanomyces Tannin Project, includingthe2001large-scalewinemakingtrial ieisadrgosSeminars,Tanunda, SA wineries andregions Dekkera/Brettanomyces conditions/non destructivemeasurement ofwhitewinecolour , andtheproduction of4-ethylphenolduringwinemaking , andtheproduction of4-ethylphenolduringwinemaking es sltdfo ifrn utain andOenology yeast isolatedfrom different Australian Saccharomyces cerevisiae essadter 1t nentoa 11Jun02 13thInternational yeasts andtheir dinner, Sydney, NSW O’Shea Award 2002 Paris, France World ofMicrobes', Congresses on'The Societies World Growers’ Day science andartofwine CSIRO Forum:the (S. SmithandSon) Seminar -Bordeaux. Working Wine With Urrbrae, SA previously Montpellier, France Coonawarra, SA quality, Urrbrae, SA 26 Jul02 18 Jul02 reported) Aug 02 57 Appendices 58 Appendices Xia Microbial inducedoxidativespoilageof bottledred wine Reducingbentoniterequirements forprotein stabilityofwine E.J. Bartowsky, D. P.W. Godden UpdateontheAWRI trialofthetechnicalperformancewineclosures E.J. Waters P.W. Godden ..Brosy .Microbial inducedoxidativespoilageofbottledred wine Xia Reducingbentoniterequirements forprotein stabilityofwine E.J. Bartowsky, D. P.W. Godden UpdateontheAWRI trialofthetechnicalperformancewineclosures E.J. Waters P.W. Godden ..Wtr Oxidationofbottledwine:therole ofascorbic acid,andbottlestorage yeasts:influence on winecompositionandflavour withalternative Winemaking P J.M. Eglintonand E.J. Waters P.J. Costello) (presented by P.A. Henschke yeasts:influenceonwinecompositionandflavour withalternative Winemaking Oxidationofbottledwine:therole ofascorbicacid,andbottlestorage and P J.M. Eglinton E.J. Waters P.J. Costello) (presented by P.A. Henschke P yeasts:influenceonwinecompositionandflavour withalternative Winemaking J.M. Eglintonand P.J. Costello) (presented by P.A. Henschke ..Gde,IL Coue eut rmteAR ra he er otbtln h oe rgt 12-14 TheRomeoBragato 9-12Sept02 XXIInternational Mouth-feel properties ofproanthocyanidins: influenceofproanthocyanidin structure Francis, J.B.Field Closures -results from theAWRI trial three yearspostbottling P.W. Godden,I.L. S. Vidal Polyphenols,‘pigmentedpolymers’andred winecolour:results from theTannin Project, M.J. Herderich P Polyphenols,pigmentedpolymersandred winecolour:results from theTannin M.J. Herderich Date Managing Whereandtowhom S-J. Bell Oxidationofbottledwine:therole ofascorbicacid,andbottlestorage P Title oftalk E.J. Waters Meeting saltspecificationsinwinemaking Microbial inducedoxidativespoilageofbottledred wine D. Xia E.J. Bartowsky, Strategiesforsuccessfulinduction ofmalolacticfermentation Thelinkbetweenvineyard andwinemakingpracticeswineprotein stability S-J. Bell P.W. Godden Managingstuckfermentation P.J. Costello E.J. Waters P.W. Godden P Staff P.R. Grbin P.J. Costello, -.Bl Managing S-J. Bell Costello, P.R. Grbin P.R. Grbin P.J. Costello, P Polyphenols,‘pigmentedpolymers’andred winecolour:results from theTannin Therole ofVA inre-starting stuckfermentation M.J. Herderich P.A. Henschke P.A. Henschke E.J. Bartowsky, E.M.C. Robinson K. Lattey, P.B. Høj, Coulter, P.J. Valente, M. Gishen,A.D. .A. Henschke .J. Costello .A. Henschke .A. Henschke .A. Henschke .A. Henschke 3 3 , G.H.Fleet , G.H.Fleet .A. Henschke 3 , G.H.Fleet 1 1 Whichbacterialstrainsandhowmanyare conductingyourMLF , Cue n oto fmuiesi ieRaso eia o 27Aug02 Roadshowseminarto: Causesandcontrol ofmousinessinwine , .. assadcnrlo osns nwn odhwsmnrt:30Aug02 Roadshowseminarto: Causesandcontrol ofmousinessinwine , P.J. 29Aug02 Roadshowseminarto: Causesandcontrol ofmousinessinwine , 3 3 , , 4 , 3 , 1 Dekkera/Brettanomyces by usingheatandenzymes Dekkera/Brettanomyces by usingheatandenzymes conditions/non-destructive measurement ofwhitewinecolour conditions/non-destructive measurement ofwhitewinecolour n fitrcin ihwn opnnsConference on and ofinteractionswithwinecomponents including the2001large-scalewinemakingtrial Project, includingthe2001large-scale winemakingtrial conditions/non-destructive measurement ofwhitewinecolour Dekkera/Brettanomyces Project, includingthe2001large-scalewinemakingtrial Botrytis Botrytis in thevineyard in thevineyard , andtheproduction of4-ethylphenol duringwinemaking , andtheproduction of4-ethylphenol duringwinemaking , andtheproduction of4-ethylphenolduringwinemaking Polyphenols, Morocco Rutherglen, Vic Pyrenees, Vic Bendigo, Vic New Zealand Conference, Christchurch, Sept 02 ..HreihPolyphenols,‘pigmentedpolymers’andred winecolour:results from theTannin Project M.J. Herderich HACCP:whatisit?DoyouneedHowdo–cheaply! Genetechnology:introductory principlesandimprovement ofgrapeandwineproduction Thelinkbetweenvineyard managementandwineprotein stability R.G. Dambergs P.A.Henschke S-J. Bell Corktaint–causes,detectionandprevention M.A. Sefton Strategiesforsuccessfulinductionofmalolacticfermentation P E.J. Bartowsky, P.J. Costello, ..FacsGaeqaiymaue itraadMra aly 12Nov02 andMurrayValley Victoria 18Nov02 20-23Oct02 Roadshowseminars: Chile2002 Vinsalud UpdateontheAWRI trialofthe technicalperformanceofwineclosures P.W. Godden ASVOSymposium– Grape qualitymeasures I.L. Francis ModerationinAustralia–policyandachievements Theeffects oftannins,anthocyanins,ethanol,andpolysaccharides C.S. Stockley I.L. Francis Vic Mornington, A.D. Coulter 18Sept02 andmeasurement Identification ofkeywineflavourcompounds Roadshowseminar: S.L. Brown Grapecolourmeasurement, Measuring desirableoakwoodcomponentsinwine G-GandNIRupdate Tannins and mouth-feel:I.L. Francis terminology Date and met R.G. Dambergs A.P. Pollnitz Thefateofagrochemical residues duringthewinemakingprocess Francis I.L. P.W. Whereandtowhom Godden P.A. yeasts:influenceonwinecomposition andflavour withalternative Winemaking Henschke J.M. Eglintonand S-J. Bell HACCP:whatisit?DoyouneedHowdo–cheaply! Thelinkbetween vineyard managementandwineprotein stability R.G. Dambergs Yarra Valley, Vic Xia Microbial inducedoxidativespoilageofbottledred wine Managingstuckfermentation E.J. Bartowsky, D. Corktaint–causes,detectionandprevention S-J. Bell 17Sept02 P.W. Polyphenols,‘pigmentedpolymers’andred winecolour:results from theTannin Godden TheuseofNIRforthemeasurement ofgrape,juiceandwinecomponents Project Tannins and Agrochemical issuesforgrapegrowers mouth-feel: andwinemakers A.P. terminology Pollnitz and met Roadshowseminar: R.G. Dambergs Title oftalk A.P.Pollnitz Francis I.L. S-J. Bell P.W. Godden Oxidation ofbottledwine:therole ofascorbicacid,andbottlestorage P yeasts:influenceonwinecompositionandflavour withalternative Winemaking J.M. Eglintonand I.L. Francis HACCP:whatisit?doyouneedhowit?-cheaply! Managing R.G. Dambergs Managingstuckfermentation Corktaint–causes,detectionandprevention S-J. Bell P.W. Godden A.P. Pollnitz Staff P .A. Henschke .A. Henschke .A. Henschke 3 , G.H.Fleet 3 (aceticacidbacteriaspoilageinwine) , (mouth-feel wheel,andtheresults ofsomerecent tanninsensorystudies) Tannins and mouth-feel:terminologyandmethodsforsensoryevaluationofred wines Grape colourmeasurement andNIR Microbial inducedoxidativespoilageofbottledred wine(aceticacidbacteriaspoilageinwine) Agrochemical issuesforgrapegrowers andwinemakers conditions/non destructivemeasurement ofwhitewinecolour Oxidation ofbottledwine:therole ofascorbicacid,andbottlestorage on red winemouth-feel Dekkera/Brettanomyces, Dekkera/Brettanomyces Saccharomyces cerevisiae Dekkera/Brettanomyces Quality ManagementSystems(ISO9000) Dekkera/Brettanomyces conditions/non destructivemeasurement ofwhitewinecolour Quality ManagementSystems(ISO9000) Botrytis in thevineyard , andtheproduction of4-ethylphenol duringwinemaking n h rdcino -typeo uigwnmkn rsnainadtsig 30Sept02 Presentation andtasting , andtheproduction of4-ethylphenol duringwinemaking , andtheproduction of4-ethylphenolduringwinemaking n h rdcino -typeo uigwnmkn SwanValley, WA and theproduction of4-ethylphenolduringwinemaking anpoen hc euevsbehz nwiewn HORsac eia 25Sept02 DHVOResearch Seminar mannoproteins whichreduce visiblehazeinwhitewine hods forsensoryevaluationofred wines hods forsensoryevaluationofred wines use ofgases Urrbrae, SA Plant Research Centre, Mildura TAFE, Vic Grapegrowers group, Council, MerbeinWine Growers Winegrape Santiago, Chile Health Conference in and Wine International Wayville, SA Show,Royal AdelaideWine provided toJudgesatthe , Burnside, SA , Burnside, The 10 Oct02 59 Appendices 60 Appendices ..FacsClu esrmns–tebsc InterwineryAnalysisGroup, 22Nov02 S.L. Br Colour measurements –thebasics I.L. Francis ..HreihPolyphenols,‘pigmentedpolymers’andred winecolour:results from theTannin Project Genetechnology:introductory principlesandimprovement ofgrapeandwineproduction Herderich M.J. P.A.Henschke HACCP:whatisit?DoyouneedHowdo–cheaply! 22Nov02 Thelinkbetweenvineyard managementand wineprotein stability R.G. Dambergs Roadshowseminars: S-J. Bell Corktaint–causes,detectionandprevention UpdateontheAWRI trialofthetechnicalperformancewineclosures M.A. Sefton P.W. Godden ThechangingfaceofmoderationinAustralia C.S. Stockley Polyphenols,‘pigmentedpolymers’andred winecolour:results from theTannin Project Herderich M.J. HACCP:whatisit?DoyouneedHowdo–cheaply! R.G. Dambergs Thelinkbetween vineyard managementandwineprotein stability Strategiesforsuccessfulinductionofmalolacticfermentation P.A. Henschke S-J. Bell Corktaint–causes,detectionandprevention M.A. Sefton A. Heinrich Henschke J.M. Eglinton Barros Lopes Lahnstein J.R. Bellon de Barros Lopes P.A. Henschke,M.A. of strain Heinrich, A. Pollnitz, V. Jiranek S. Dillon Date Whereandtowhom Title oftalk 20Nov02 Roadshowseminars: UpdateontheAWRI trialofthetechnicalperformancewineclosures P.W. Godden Staff ..FacsCaatrsn pcfcatignypoete frdwns utaainAscain 11Dec02 AustralasianAssociation Characterising specificastringencyproperties ofred wines I.L. Francis Barros Lopes C. Hillier Eglinton, A.P. Pollnitz, Ferguson L. Dulau Bartowsky, P.B. Høj, Fleet andeffect onwinecomposition E.J. Bartowsky, G.H. eBro oe,mnortiswihrdc iil aiesi ht ie Discoverymeeting, K.S. Howell mannoproteins whichreduce visiblehazinessinwhitewine P.B. Høj,E.J.Waters de Barros Lopes, 3 , P.A. Henschke own .. Canyeaststrainaffect colourandphenoliccontentofShirazwine? 5 , E.J. 7 6 , P.A. , M.A.de , G.Currie 1 1 , M.de , K. .. Hybridwineyeastswithuniquefermentation characteristics , J.M. .. Investigationsintothe mechanism ofactionandbiologicalrole of , M.A. Yeast straindynamicsduringmixedculture winefermentation , . Comparativeproteomics: identifyingnovelproteins inthewinemakingstrain , J. .. Usingmetabolomicstoinvestigatecellularmetabolicchangesinan overproducing , A.J. 6 , Tannins andmouth-feel:terminologymethodsforsensoryevaluationofred wines Grape colourmeasurement andNIR Agrochemical issuesforgrapegrowers andwinemakers Salty wine conditions/non destructivemeasurement ofwhitewinecolour Oxidation ofbottledwine:therole ofascorbicacid,andbottlestorage Dekkera/Brettanomyces wines (mouth-feelwheel,andtheresults ofsomerecent tanninsensorystudies) Tannins andmouth-feel:terminologymethodsfor sensoryevaluationofred Grape colourmeasurement andNIR (acetic acidbacteriaspoilageinwine) Microbial inducedoxidativespoilageofbottledred wine Agrochemical issuesforgrapegrowers andwinemakers conditions/non destructivemeasurement ofwhitewinecolour Oxidation ofbottledwine:therole ofascorbicacid,andbottlestorage Dekkera/Brettanomyces, acaoye cerevisiae Saccharomyces (mouth-feel wheel,andtheresults ofsomerecent tanninsensory studies) acaoye cerevisiae Saccharomyces n h rdcino -typeo uigwnmkn Margaret River, WA , andtheproduction of4-ethylphenolduringwinemaking n h rdcino -typeo uigwnmkn Great WA Southern and theproduction of4-ethylphenolduringwinemaking S. cerevisiae lb HealesvilleVic Club, Seminar, RACVCountry Interwinery Group Review Adelaide, SA 02 Nov 20 Drug Conference, 2002 APSADAlcoholand Heron IslandQld 5th Scientificmeeting, for ChemosensoryScience 2 Melbourne, Vic Melbourne, nd es rdcsad27-29Nov02 Yeast Products and M.A. Gishen P.W. Godden ..Hnck Overviewofwinemicrobiology/biotechnology projects R.E. Day AWRI i P.A. Henschke P.W. Godden Herderich Håkansson, M.J. Asenstorfer, A.E. G. Harvey Iland Dawson ..Dmeg esrn rp oorCC il eia,28May03 CRCVFieldSeminar, Measuring grapecolour R.G. Dambergs M. Henry Massis, A.Catalano, M. Holdstock,A. L. Janik Beyondtheidea:wineindustry’s approach tobridgingthegapbetween Pretorius, R.E.Day P.B. Høj,I.S. Potentialcardiovascular benefitsofmoderatewineconsumption Therole oftheAWRI IndustryServicesteam M.J. Herderich tanninandcolourspecification Winegrape microbiology/biotechnology projects: Wine microbial P.W. Godden Measuring aroma compoundsinwine P.A. Henschke I.L. Francis M.A. Sefton Date Whereandtowhom Title oftalk Y 28Jan03 from Orlando Winemakers P.W. Godden Francis, P.B. Høj Cozzolino, I.L. Dambergs, D. Sulfidebiochemistry:microbiological originandcontrol of P.A. Henschke Staff ..PeoisAgnrlitouto oTeAsrla ieRsac nttt & al,U eir 26Mar03 E&JGallo,USsenior Sefton, E.J.W I.L. Francis,M.A. role ofascorbicacid,andbottlestorageconditions M.J. Kwiatkowski, wine: bottled of Oxidation G.K. Skouroumounis, Research Ageneralintroduction Institute toTheAustralianWine I.S. Pretorius Yeast-mediated formationofanthocyanin-derivedpigmentsinred wine M.J. Herderich I.S. Pretorius M. Gishen P.B. Høj D. Cozzolinoand Boehm, I.LFrancis, W.U. Cynkar, D. Dambergs, L.Janik, ..FacsCmo iefut n o oaodte lcwo ieaes 16Apr03 BlackwoodWinemakers 4Apr03 UniversityofAuckland, Commonwinefaultsandhowtoavoidthem M. Gishen Yeast mediatedformationofanthocyanin-derivedpigmentsinred wine I.L. Francis M.J. Herderich . Hayasaka 1 , 8 , R.G. , D. Buick, FOSS ‘WineScan’—calibration developmentforwineanalysis FOSS‘WineScan’—calibration , D.Buick, 9 2 11 , P.B. Høj, .. epnet F & eiwWARDRve 21Mar03 WFA R&DReview ResponsetoWFA R&DReview , P.J. , M.Gishen 10 , R.G. NIR in the wine industry Wine usersgroup seminar Wine NIRinthewineindustry , R.G. n Legalmetrology inthe wineindustry and , R. Screening for potential wine pigments in red wine using tandem mass spectrometry The 19th conference of 2-6 Feb 03 Feb 2-6 conference19th of The Screening forpotentialwinepigmentsinred wineusingtandemmassspectrometry R. , .. rp n ieqaiyatiue n esrmnsVcoinWn nuty 30–31Jan03 Industry Wine Victorian Grapeandwinequality-attributesmeasurements , P.G. aters 2 Dekkera/Brettanomyces uigwnmkn tsigadpeetto)Saltramwinery, Barossa during winemaking(tastingandpresentation) modulation ofwinesensoryproperties Wy Dekkera/Brettanomyces sulfur-compounds inwinemaking during winemaking (tasting and presentation) Blass Wines BlassWines during winemaking(tastingandpresentation) the ideaanditstransformationtocommercial outcomes cnd’cluei usi feclec PowerhouseMuseum, Warren Centre, ‘can-do’ culture inpursuitofexcellence The importanceofindustry/researcher communicationasadriverfor ndustry support and development activities – the role of the Industry Services team A gr A ndustry supportanddevelopmentactivities–therole oftheIndustryServicesteam n h rdcino eaiesnoycmons eigrBassafa 31Jan03 BeringerBlassstaff at and theproduction ofnegativesensorycompounds n h rdcino eaiesnoycmons ieaesa eigr31Jan03 atBeringer Winemakers and theproduction ofnegativesensorycompounds non-destructive measurement ofwhitewinecolour, the Valley, SA winemakers, Urrbrae,SA Victoria Lorne, Spectrometry heldin Society forMass Australian andNewZealand Mildura, Vic Association's noain eodthe beyond Innovation: Nashville, USA University MedicalCentre, pharmacology Grand roundsinclinical Sunshine, Vic Convention Centre, University Victoria outlook quality andindustry Urrbrae, SA production team,AWRI, New Orleans,USA American ChemicalSociety, of redwinepigments Uncovering themysteries Centre, AdelaideSA Wine Committee, National New Zealand conference, Taupo, at FOSSDirections 2003 idea Blackwood, SA and Brewers Club, New Zealand Ultimo, NSW ndham, RowlandFlat,SA u fTa 17Mar03 oup ofThai seminar The conference Grape Vanderbilt 3 Apr03 1 Apr03 23 Mar03 28–29 May03 61 Appendices 62 Appendices ..Dmeg ISi h ieidsr rpgoesad18-22 Aug 02 Grapegrowers and Date presented towinemakers Location S-J. Bell presented towinemakers NIRSinthewineindustry ofhazes/depositsandusemicroscope Isolation R.G. Dambergs 16Aug02 Laboratoryanalysisandqualitymanagement Introduction toAWRI technicalmanualWebsite Date A.D. Coulter E.M.C. Robinson Title ofpresentation Roadshowworkshop Mark Gishen Godden Tasting ofsimulatedfaultywines M. Gishen,P.W. Whereandtowhom Trouble free winemaking–causesandprevention ofcommonwine instabilities E.M.C. Robinson, A.D. Coulter, Isolationofhazes/depositsandusemicroscope P.W. Godden Laboratoryanalysisandqualitymanagement 3Jul02 Introduction toAWRI technicalmanualWebsite A.D. Coulter E.M.C. Robinson Campus,UrrbraeSA Roadshowworkshop M. Gishen Godden Tasting ofsimulatedfaultywines M. Gishen,P.W. Trouble free winemaking–causesandprevention ofcommonwineinstabilities E.M.C. Robinson, A.D. Coulter, Plant Industry, Waite P.W. Godden 29Jun Title oftalk 1stFEMSCongress of Staff Workshops Novelapproaches towinemaking–tailoringwineyeastforfuture challenges 12-13 Jun 0 FormazionePermanente I.S. Pretorius Sensoryanalysis Present andfuture developmentsinalcoholicfermentationmanagementAustralia flavourandaroma. Whatcanwemeasure Wine now? Flavourcompoundsinwineandgrapes P.A. Henschke I.L. Francis M.A. Sefton P.B. Høj Metabolicprofiling Fermentationtechnologies Lopes M.A. deBarros P.A. Henschke Staff ..Cutr ...Tsigo iuae alywnsBnor aps Oct02 16-18 BundooraCampus, Grapegrowers and 13Dec02 TheAustralianWine S-J. Bell Workshop tosummariseGWRDC-fundedresearch outcomesforwineandhealth ofhazes/depositsandusemicroscope Isolation C.S. Stockley Laboratoryanalysisandqualitymanagement 29,30,31 Introduction toAWRI technical manualWebsite A.D. Coulter Trobe La University’s E.M.C. Robinson M. Gishen Gishen, P.W. Godden Trouble free winemaking–causesandprevention ofcommonwine instabilities Robinson, M. Tasting ofsimulatedfaultywines A.D. Coulter, E.M.C. P.W. Godden S-J. Bell NIRSinthewineindustry R.G. Dambergs ult aaeetSses–IO90 winem Research toPractice Quality ManagementSystems–ISO9002 What bugisthat? What bugisthat? yegsi neato ihvtmnEUrrbrae, SA Research Institute, Research toPractice inYarra Winemakers synergistic interactionwithvitaminE Potential cardioprotective activitiesofwinecomponentsbasedon What bugisthat? Research toPractice Quality ManagementSystems–ISO9002 TM TM ierp ult aaeetwrso ra oten6-7 Jan03 Great Southern QualityManagementworkshop Winegrape rpvn urto aaore A17-18Oct02 Naracoorte, SA 4-5Sept02 Creek, Langhorne SA Grapevine Nutrition Grapevine Nutrition wnVle,W 9-10Jan03 Swan Valley, WA Margaret River, WA Great and Southern Conegliano, Italy ConferenceVitivinicola ntblt problems instability wine troubleshooting and Tas, and grapegrowers, Hobart, wine instabilityproblems and troubleshooting Trouble freewinemaking Coonawarra, SA, and grapegrowers, Ljubijana, Slovenia to3Jul03 European Microbiologists, vine topalate. flavour andaromafrom First workshoponwine wine industry members oftheVictorian 24-25Sept02 Peninsula, Vic Valley andMornington Vic Mildura, rul rewinemaking free Trouble akers inSwanValley, CSIRO 30 May03 Sept 02 3 .Dlo,EJ Canyeaststrainaffect colourand L. Dulau phenoliccontentofShirazwine? Bartowsky, P.B. Høj, S. Dillon,E.J. culture winefermentationsand effect onwinecomposition Fleet, P.A. Yeast straindynamicsduringmixed Henschke wineyeast? –analternative Bartowsky, G.H. K.S. Howell,E.J. P.A. Henschke J.E. Eglinton, de Barros Lopes overproducing strainof P.A. Henschke,M.A. cellularmetabolicchangesinan Heinrich, A.Pollnitz, D. Torrea, C.Ancin Henschke uhrTteo otrOgnsto n oainDate Organisationandlocation USA Wisconsin, SeventhSymposiumonLacticAcidBacteria: 30Jul-4Aug02 Assessmentofimportance ofbacterial E.J. Bartowsky, J.M. Title ofposter 2002Yeast GeneticsandMolecularBiologyMeeting,Madison, 27Jul–1A actionandbiologicalrole of P.B. Høj,E.J.Waters Investigationintothemechanismof UnionofMicrobiological International Societies,Paris,France de Barros Lopes, S.L. Brown, M.A. culture wine fermentationandeffect ofwinecomposition Fleet, P.A. Yeast straindynamicsduringmixed Henschke Bartowsky, G.H. K.S. Howell,E.J. Author Posters presented ..PeoisRme1p inducesFLO11expression MolecularBiology(ICGMB)Göteborg,Sweden I.S. Pretorius de Barros Lopes P.A. Henschke,M.A. Heinrich, A.P. Pollnitz,genes andwineflavour ..Dne h osmto f h 9hRylAsrla hmclIsiue 6-11Jul03 The 19thRoyalAustralianChemicalInstitute 21 Investigatingthelinkbetweenyeast Theconsumption of J.M. Eglinton,A.J. M.A. Daniel NIRinstrumentsfordeterminationof colourinred grapes Cynkar, I.L.Francis Dambergs, W.U. Calibrationtransferbetweendifferent Gishen, R.G. D. Cozzolino,M. red grapesusingadiode arrayinstrument Cynkar Dambergs, W.U. DeterminationofcolourandpH Apr 03 Gishen, R.G. D. Cozzolino,M. Cynkar, M.J. Herderich infrared transmission (NIT)spectroscopy R.G. Dambergs,W.U. duringred winefermentationbynear Gishen, Parker,M. Measurement ofphenoliccompounds Kwiatkowski, M. D. Cozzolino,M.J. M. Gishen L.J. Janik,I.L.Francis, NIRSConference, 11thInternational Spain qualityparameters error withNIRSanalysisofwinegrape Cynkar, M.B.Esler, Cozzolino, W.U. Strategiestominimisematrix-related R.G. Dambergs,D. D. Torrea, C.Ancin ..Elno,AJ Uigmtblmc oivsiae2 whitewineanditssensorycontribution Usingmetabolomicstoinvestigate J.E. Eglinton,A.J. P.A. Henschke McCarthy, I.L.Francis, 5 , P.A. 12 12 acaoye bayanus Saccharomyces cerevisiae Saccharomyces reduce visiblehazinessinwine S. cerevisiae production byrecombinant yeast Influence offlocculationonethanol in wineyeast Factors affecting fructoseconsumption Maltotriose transportinyeast shuttle inyeast Ras regulates thecarnitine response element through an11bpRme1p -aacnn nwn OrganicConference (19RACI-OC),CumberlandConference ß-damascenone inwine requiring yeaststrains inoculated withtwodifferent nitrogen during fermentationofgrapemust Production ofvolatilemetabolites with theirprecursor aminoacids with different yeaststrains:relationship Biogenic aminesinwinesobtained strain ondiacetylproduction inred and mannoproteins that Discovery Resort, Lorne Victoria Resort, Lorne Metabolism andApplications nd st Australian Conference onYeast: nentoa ofrneo es eeisad7-12Jul03 ConferenceInternational onYeast Geneticsand , Melbourne, Vic , Melbourne, , EgmondaanZee,TheNetherlands Products and Genetics 1-5 Sept02 , 27-29 Nov02 ug 02 63 Appendices 64 Appendices aaeeto omnwn ntblte . or P.W. Godden P.W. P.A. Godden Henschke I.L.Francis P.W. Godden A.D.Coulter P.W. 1.5 hours Godden E.J.Waters 1hour M.A. deBarros Lopes 2 1hour 2 P.W. Godden 8 hours 1 3 1 M. Gishen Francis I.L. 2 hours wine closure - An evaluationofthetechnicalperformance ofvarioustypes during winemaking,includingatasting ofwinescontainingkeyspoilagecompounds C.S. StockleyandP.B. Høj Dekkera/Brettanomyces 1 2 management ofcommonwineinstabilities P.A. Henschke Trouble free winemaking–theidentificationand C.S. Stockley Defining andmeasuringgrapewinequality Approx. 50hours P.W. Godden Australian wineindustryandstructure E.J. Bartowsky LaTrobe University Industry 3002WT BiotechnologyintheFoodandWine 1 3009WT Advancedsensorypractice 3009WT Advancedsensorypractice 2 1005/3113 Winemaking 1 2580 Stabilisationandclarification 3 2580 Stabilisationandclarification 3005WT Grapeindustrypracticepolicyandcommunication andSociety) (final yearwinemarketingstudentsaspartofthecourseentitledWine 2001WT forundergraduatesand7030WTpostgraduates packagingandqualitymanagement 1958 Wine The Universityof Adelaide 1 2003 –Semester Date MMED 3921Industrialandpharmaceuticalmicrobiology Flinders University 2022WT Sensorystudies packagingandqualitymanagement 3003WT Wine 3045WT Advancesinoenology The Universityof Adelaide 2002 –Semester2 Subject Teaching responsibilitiesofInstitutestaffduring2002/03 APPENDIX 2. Activity 12 UniversidadPublicadeNavarra 11 FOSSPacific 10 NationalStandards Commission 23-26Jul02 Company 9 Hardy Wine 21Feb03 8 now, DavidsonViticulture 7 FlindersUniversity 6 SchoolofBotany, UniversityofMelbourne 5 Lallemand 4 FieldConsultingServices 3 DepartmentofFoodScience,UniversityNewSouthWales Ricard2 Pernod Improvement, SouthAustralianVine Rieslingclonalselectiontrial.Participatedasapanel TheUniversityofAdelaide 1 SchoolofAgriculture andWine, P.W. Godden K.A. Lattey Coulter, I.L.Francis, T.A. Weber, A.D. AssessmentCourse,Waite AdvancedWine Campus,UrrbraeSA Robinson, M.Gishen, P.W. Godden,E.M.C. Staff Other activities Results to42monthspostbottling , andtheproduction ofnegativesensorycompounds member duringapublictasting o flcue Institutestaff of lectures No. . or P.W. Godden 1.5 hours P.B. Høj M.J. Herderich P.A. Henschke P.W. Godden Francis I.L. M.A. deBarros Lopes 1 5 4 1 2 1 APPENDIX 3. Graduate and Honours student supervision responsibilities of Institute staff for 2002/03

Student Supervisor/s Source of funds

PhD S.L. Brown E.J. Waters, M.A. de Barros Lopes, P.B. Høj GWRDC F. Carrau P.A. Henschke, E. Dellacassa1 University of the Republic of Uruguay staff member M.A. Daniel G.M. Elsey, M.A. Sefton, M. Perkins2 CRCV M. de Sa M.J. Herderich, I.L. Francis, A.P. Pollnitz CRCV J.M. Eglinton P.A. Henschke, P. Langridge3 Institute staff J. Gardner V. Jiranek3, M.A. de Barros Lopes School of Agriculture and Wine – The University of Adelaide W. Greenrod M. Fenech4, M. Abbey4, P. Burcham5, C.S. Stockley GWRDC A. Grimaldi V. Jiranek3, E.J. Bartowsky School of Agriculture and Wine – The University of Adelaide /GWRDC C. Guirado G.K. Skouroumounis, E.J. Waters Visiting French Postgraduate student (until 25 October 2002) A.J. Heinrich M.A. de Barros Lopes, V. Jiranek3 CRCV K.S. Howell P.A. Henschke, E.J. Bartowsky, G. Fleet6, M.A. de Barros Lopes University of NSW/GWRDC A. Janusz G.M. Elsey, M.A. Sefton, M. Perkins2 CRCV C. McBryde V. Jiranek3, M.A. de Barros Lopes School of Agriculture and Wine – The University of Adelaide O.J. Macintyre C. Colby7, B. O’Neill7, E.J. Waters, I.S Pretorius School of Chemical Engineering – The University of Adelaide/GWRDC R. Muhlack C. Colby7, B. O’Neill7, E.J. Waters, P.B. Høj, A. Lim8 School of Chemical Engineering – The University of Adelaide/Hardy Wine Company/GWRDC C.J. Puglisi G.M. Elsey, M.A. Sefton, R. Prager2 CRCV R. Ristic P.J. Iland3,9, I.L. Francis, M.J. Herderich GWRDC H.E. Smyth I.L. Francis, M.A. Sefton, M.J. Herderich GWRDC K.L. Wilkinson G.M. Elsey, M.A. Sefton, R. Prager2 GWRDC

Masters M. Astorga M.A. de Barros Lopes, V. Jiranek3 School of Agriculture and Wine, The University of Adelaide D. Coates E.J. Bartowsky School of Agriculture and Wine, The University of Adelaide

Honours R.C. Brown G.M. Elsey, D. Taylor3 Flinders University M. Caldersmith A.P. Pollnitz, S.J. Clarke3 School of Agriculture and Wine – The University of Adelaide J.S. Crossman G.M. Elsey Flinders University J. Guzinski M.A. de Barros Lopes, Carolyn Leach10 Department of Genetics – The University of Adelaide N.R. Sleep G.M. Elsey, D.L. Capone Flinders University

Industry placement B. Winstone, M.A. de Barros Lopes Department of Biotechnology, Flinders University Jan 03

Theses completed - Honours/PhD

Student Hon/PhD Title of thesis Supervisors

J. Cartwright Hons The effect of interspecific Saccharomyces hybrids on the sensory and M.A. de Barros Lopes, chemical profiles of wine A.J. Yap3 M. Fettke Hons The role of amino acids in wine yeast and lactic acid bacteria compatibility P.A. Henschke, E.J. Bartowsky, P.J. Costello K. Poole PhD Increasing the fermentation reliability of wine yeasts M.A. de Barros Lopes, by enabling proline utilisation V. Jiranek3 K. van Leeuwen Hons The development of stable isotope dilution assays for the G.M. Esley quantification of the important aroma compounds citronellol, 4-vinylguaiacol and 4-vinylphenol in wines

1 University of Republic of Uruguay 2 Flinders University 3 The University of Adelaide School of Agriculture and Wine 4 CSIRO Health Sciences and Nutrition 5 The University of Adelaide Department of Clinical and Experimental Pharmacology 6 University of New South Wales 7 The University of Adelaide School of Chemical Engineering 8 Hardy Wine Company 9 now, Davidson Viticulture 10 The University of Adelaide Department of Genetics Appendices 65 66 Appendices 0 gitn . eshe .Winemaking propertiesof P. Henschke, J.; Eglinton, 2002. 81; NZGrapegrower Winemaker (465): Aust. Furtherinsightsintothecardioprotectivepropertiesofwine. C. Stockley, 707 NZGrapegrower Winem 706 Aust. To labelornottoforallergens-thelatestquestionwinemakers. NZ Grapegrower R. Winemaker (4 O'Hehir, Aust. Powdery mildewandgrapewinequality. C.; E.S. Stockley, Scott, A.J.; Markides, I.L.; Francis, B.E.; Stummer, 705 Objectivemeasuresofgrapea P.B. Høj, R.S.; Johnstone, A.; Kambouris, 704 I.L.; Francis, M.B.; Esler, R.G.; Dambergs, P.G.; Iland, M.; Gishen, Someapplicationsofanalysesv M.A. Sefton, 703 G.K.; Skouroumounis, H.; McLean, S.; Franke, J.I.; Campbell, D.L; Capone, A.P.; Pollnitz, 702 Willia R.J.; Blair, Management ofmalolacticfermentation-wineflavourmanipulation. P.A. Henschke, P.J.; Costello, E.J.; Bartowsky, Genet M.A. deBarrosLopes, P.B.; Høj, P.A.; 701 Henschke, P; Langridge, A.J.; Yap, A.J.; Heinrich, A.P.; Pollnitz, J.R.; Bellon, J.M.; Eglinton, 700 2 atwk,E yoyeadwnmkn.As.N rpgoe ieae 43) 0-0;2003. 101-104; NZGrapegrower Winemaker (473a): Aust. Lysozyme andwinemaking. E. Bartowsky, 728 Winequalitygradingbynearinfraredspectroscopy M. Gishen, M.B.; Esler, I.L.; Francis, N.; Salinityeffectsonvinesandwines. Schumacher, E. A.; Robinson, Kambouris, P.; R.G.; Valente, Dambergs, L.; Francis, P.; Godden, P.R.; Clingeleffer, D.H.; Blackmore, R.R.; Walker, 724 Effects ofvariety andregiononnearinf D.R. Boehm, W.U.; Cynkar, 723 A.; Kambouris, R.G.; Dambergs, I.L.; Francis, M.; Gishen, M.B.; Esler, butteryaromaperceptioninwines predictablefromthediacetylconcentr Is P. Henschke, J.R.; Bellon, 722 L.; Francis, E.J.; Bartowsky, theimportanceofindustry-researchercommunicationandenhancedR&D investm Beyondtheidea: R.E. Day, 721 I.S.; Pretorius, P.B.; a Høj, linalool, nerol, Quantitativeanalysisofgeraniol, Mic M.A. Appl. Sefton, Lett. A.P.; Pollnitz, G.K.; Skouroumounis, Spoilageofbottledredwinebyaceticacidbacteria. 720 D.L.; Capone, P.A. Henschke, D.; G.H.; SejerPedersen, Fleet, R.L.; Gibson, D.; The mouth-feelpropertiesofgrapeand Xia, E.J.; E.J. Bartowsky, Waters, V.; Cheynier, R.; Gawel, 719 M.; Kwiatkowski, N.; Marnet, S.; Guyot, 718 L.; Francis, S.; Vidal, Combined heatandproteolyticenzymetreatmentofwhitewinesr E.J. Waters, M.J.; Kwiatkowski, 717 K.S.; P.B.; Adams, Høj, K.F.; acasestudyontwovarieties attwowiner Pocock, Leadinwine: T.H. Lee, C.D.; Osborn, B.L.; Gulson, K.G.; Tiller, L.H.; 716 Smith, C.S.; Stockley, 715 2 aaaa . aNmr,K;Blok .. alr .. oliz ..Application ofstirbarsorptiveextractionforwineanalysis A.P. Pollnitz, 2003. R.L.; Taylor, 58-62; G.A.; 18(1): Baldock, J. K.; NZ Wine Ind. MacNamara, Aust. Y.; Producingwinestylesfor overseasmarkets. Hayasaka, J. Hughes, P.; Godden, M.; Tetrahed Structuresandcolourpropertiesofnewredwinepigments. Gishen, M. R.; Herderich, Day, M.; deSa, Y.; Hayasaka, 727 K.; Pardon, A.E.; Hakansson, 726 725 The effectofpowderymildewinfectiongrapeberriesonjuiceandwine E.S. Scott, A.J.; Markides, I.L.; Francis, B.E.; Stummer, Characterization ofproanthocyanidinsingrapeseedsusingelectr S. Vidal, M.J.; Herderich, V.; Cheynier, 714 E.J.; 2002. Waters, Y.; 15; Hayasaka, 9, News 13(6): Near infraredspectroscopyat NIR. The Australian B. Wine ResearchInstitute(Part 1). Dambergs, M.; Gishen, 713 Asurveyofindustrialstrains of V. 712 Jiranek, P.A.; Henschke, F.J.; Meneses, precursorsandbiosynthesisofthecausativeN-heterocycles2-ethyltetra 711 Mousy off-flavorofwine: P.A. Henschke, P.J.; Costello, butteryaromaperceptioninwinespredictablefromdiacetylconcentr Is P.A. Henschke, J.R.; Bellon, 710 I.L.; Francis, E.J.; J. Bartowsky, Aust. Permeation of2,4,6-trichloroanisolethrough corkclosuresinwinebottles. M.A. Sefton, G.K.; Skouroumounis, D.L.; Capone, 709 708 9 Kitosi .. egZK;Wtr,EJ;Gde,PW;Dy .. lre .. edrc,MJ oyhnl,pigmentedpolymersandre Polyphenols, M.J. Herderich, S.J.; Clarke, C.J.; Day, P.W.; Godden, E.J.; Waters, Peng Z.K.; M.J.; Kwiatkowski, 699 Willi R.J.; Blair, Relationshipbetweenseedcompositionandgrapewinequality. P.G. Iland, R.; Gawel, I.L.; Francis, R.; Ristic, Characterising mouth-feelproper E.J. Waters, M.J.; 698 Kwiatkowski, S.; Guyot, V.; Cheynier, S.; Vidal, P.G.; Iland, R.; Gawel, I.L.; Francis, Anevaluation ofthetechn E.M.C. Robinson, P.B.; 697 Høj, P.J.; Valente, A.D.; Coulter, M.; J.B.F.; Gishen, Field, I.L.; Francis, P.W.; Godden, P.B., Høj, P.; Williams, R.J.; Blair, Producinguniquewinestylesforoverseasmarkets. J.B. Hughes, P.W.; Godden, M.; 696 Gishen, R.E.; Day, The effectsofsalinityonvinesandwines E. Robinson, P.; Valente, L.; Francis, P.; Godden, P.; Clingeleffer, 695 D.; 2000. Blackmore, 54–56; R.; Walker, 15: J. NZ Wine Ind. Aust. ofan Investigation ‘electronic nose’. W. Cynkar, L.; Francis, 694 653a Institutestaffpublications2002/03 APPENDIX 4. 06;2002. 60–61; 188–194. 2002: S Wine Industry Technical ConferenceInc.; Adelaide, 7–11October2001; Proceedingsoftheeleventh Australian wineindustry technicalconference; eds. P.B., Høj, P.; Williams, 162–164. 2002: Australian Wine Industry 7–11Octo TechnicalAdelaide: ConferenceInc.; Proceedingsoftheeleventh Australian wineindustrytechnicalconference; eds. P.B., Høj, P.; Williams, R.J.; Blair, to wine. 157–161. 2002: Conference Inc.; Australian Win Adelaide: SA. Adelaide, 7-11October2001; Proceedings oftheeleventh Australian wineindustrytechnicalconference; 152–156. 2002: Australian Wine Industry Technical Adelaide: ConferenceInc.; SA. Adelaide, Proceedingsoftheeleventh Australian wineindustrytechnicalconferen eds. P.B., Høj, P.; Williams, R.J.; Blair, improvement ofwine. 150–151. 2002: Australian Wine Industry Technical Adelaide: ConferenceInc.; SA. Adelaide, October 2001; Proceedings oftheeleventh Australian wineindustry eds. P.B., Høj, P.; Williams, R.J.; Blair, the 2001largescalewinemakingtrial. 4-5;2003. 948-955; 1 2002: NIR Publications; UK: West Sussex, proceedingsofthetenthinternational conference. (eds.) Nearinfraredspectroscopy: R.K., 249-253. 2002: NIR Publications; UK: West Sussex, international conference. procee (eds.) Nearinfraredspectroscopy: R.K., Cho, A.M.C.; Davies, spectroscopic analysisofqualityparametersinredwinegrapes. 2003. 49–53; 47, 18(2): 2003. 22–23; 18–20, 18(2): J. WineInd. N.Z. Aust. culture inpursuitofexcellence. 2003. 517–522; 375: Chem. Bioanal. Anal. 2003. 83:564–573; Food Agric. Sci. in awine-like mediumJ. 2003. 56–63; 9: Grape Wine Res. J. Aust. protein contentwithoutdetrimentaleffect. 2003. 47–55; 9: Res. Grape Wine 2003. 28–39; 9: Grape Wine Res. J. Aust. properties ofChardonnaywines. ai omn asSeto.1:91;2003. 9–16; 17: MassSpectrom. Rapid Commun. 2002. 310–321; 108: Brew. Inst. J. utilisation. and2-acetyl-1-pyrrolineby 2-acetyltetrahydropyridine, 2002. 180–185; ofrneIc;20:145–149. 2002: Conference Inc.; Australian Win Adelaide: SA. Adelaide, 7–11October2001; Proceedings oftheeleventh Australian wineindustrytechnicalconference; 123–127. 2002: S Wine Industry Technical ConferenceInc.; Adelaide, 7–11October2001; Proceedingsoftheeleventh Australian wineindustrytechnicalconference; eds. P.B., Høj, P.; Williams, 44–52. 2002: Australian Wine Industry Technical Adelaide: ConferenceInc.; 7 SA. Proceedingsoftheeleventh Australian wineindustrytechnicalconference; eds. P.B., Høj, P.; Williams, R.J.; Blair, bottle closures. Australian Wine Industry Technica Adelaide: SA. Adelaide, 7–11October2001; eleventh Australian wineindustrytechnicalconference; 2002. 20–22; 16–18, Saccharomyces bayanus Lactobacillus hilgardii Saccharomyces cerevisiae emnaindmnne ut ZGaerwrWnmkr(6) 75;2002. 47–51; NZGrapegrower Winemaker (466): Aust. - fermentationdominance. S 07.J gi.Fo hm 0 0978;2002. 7079–7087; 50: Food Chem. Agric. J. DSM 20176. reveals numerousalteredpatternsofmaltoseandsucrose rp ieRs :1619 2002. 196–199; 8: Res. Grape Wine hydropyridine, to?As.J rp ieRs 8: Res. GrapeWine J. ation? Aust. ams,P.;Høj,P.B.,eds. d.Proceedingsofthe eds. composition andonsensory kr(6) 25,57–58, 52–54, aker (463): nl iaa.Ce.375: Chem. Bioanal. Anal. . to?As.NZ ieId J. Ind. Wine N.Z. ation? Aust. 1 coe 01 Adelaide, –11 October2001; ospray massspectrometry. s . ø,PB,eds. P.B., Høj, P.; ms, ofrneIc;20:27–30. 2002: l ConferenceInc.; ehia ofrne 7–11 technical conference; iso e ie.Bar R.J.; Blair, ties ofredwines. ut ii.64:11–14, 6(4): Vitic. Aust. . ic approachestothe ul I 6 0-2;2003. 200-227; OIV76: Bull. ical performanceofwine e nSuhAsrla ut J. Aust. ies inSouth Australia. o et 4 8749;2003. 4887-4891; 44: ron Lett. e 7–11October2001; ce; e 01 dlie SA. Adelaide, ber 2001; nd dings ofthetenth oil 6 0–1;2003. 307–314; 36: robiol. ent asdriversfora ‘can-do’ 87-189. educes hazeforming .Aead:Australian Adelaide: A. .Aead:Australian Adelaide: A. olatile flavourcompounds ieclu:resultsof d winecolour: e Industry Technical rared reflectance dwn ult.Bar R.J.; Blair, nd winequality. ais ... Cho, A.M.C.; Davies, . e Industry Technical α appleproanthocyanidins 4:6–9 2 4 2002. 74; 72, 68–69, 64): -terpineol inwine. alHnck C X X X C X X X X C X X X X C =denotesholderofChair C X Elizabeth Waters X Randell Taylor X X X Creina Stockley X X X C Mark Sefton X Sakkie Pretorius Alan Pollnitz X Ken Pocock X X X Ingrid Oats X Hans Muhlack X X Matthew Holdstock Peter X Markus Herderich X Paul Henschke Yoji Hayasaka X Peter Godden X Holger Gockowiak Mark Gishen Leigh Francis Peter Eichinger Jeff Eglinton X X X Rachel Edwards Maria deSa Miguel deBarros Lopes Catherine Daniel Adrian Coulter Amanda Cook Rae Blair Sally-JeanEveline Bartowsky X Høj CXXXXX BellXXXXX tf ebrMngmn Rsac nutySrie omncto nltclSrieIfrainBoaeyOcptoa elhStaff Cod Occupational Health Biosafety Information AnalyticalService Communication IndustryServices Research Management Staff member InstituteCommittees APPENDIX 5. P.B. Varietal differentiationofgrapejuicesbyprotein Høj, I.S.; Pretorius, E.J.; Waters, K.F.; Pocock, G.A.; Baldock, Y.; Hayasaka, Differentiation of Australian wineisolatesof P.A. Henschke, 734 J.M.; McCarthy, E.J.; Bartowsky, Standardisedmethodologyfortestingmalolacticbacteriaandwineyeastcompatibili A.J. 733 Markides, P.A.; Henschke, P.J.; Costello, situmeasurementofwhitewineabsorbanceinclearand In E.J. Waters, R.; Gawel, M.A.; Sefton, 732 M.; Kwiatkowski, G.K.; The Skouroumounis, useofnearinfraredspect M. Gishen, P.B.; Høj, I.L.; Francis, A.; Kambouris, W.U.; Cynkar, M.B.; Esler, D.; 731 Cozzolino, R.G.; Dambergs, (2003) Winemaking propertiesof P. 730 Henschke, J.; Eglinton, 729 .1() 73;2003. 27-31; 18(3): J. 2003. 122-126; 9(2): Res. GrapeWine J. Aust. 2003. 127-137; 9(2): 2003 138-148; 9(2): Grape Wine Res. J. Aust. using amodifiedlaboratoryspectrophotometer. 2003. 69-76; NZGrapegrower Winemaker (473a): Aust. measurement. 18-20. (473a): Grapegrower Winemaker dioySern teigSern teigTcnlg n aeyNegotiation andSafety Technology Steering Steering Steering Steering Advisory Saccharomyces bayanus X XCCX iiilosrain fteptnilfrrdwnmkn.As.NZ Aust. -initial observations ofthepotentialforredwinemaking. Oenococcus oeni X X X XXX using randomamplifiedpolymorphicDNA(RAPD). XX X X X fingerprinting. Aust. NZ Wine Ind. NZWine Aust. fingerprinting. ty. Aust. J. Grape Wine Res. GrapeWine J. Aust. ty. roscopy forgrapequality X X in colouredbottles X X e 67 Appendices 68 Institute Staff Second row Fourth row Front row Third row Third Absent The staff of The Australian Wine Research Institute Jane McCarthy, Emma-KateWhite Kate Beames,BelindaBramley, Jeff Eglinton,MatthewHoldstock,KateHowell,DanielleLeedham, Mark Sefton,MatthewCream, Greg Ruediger, PeterGodden Tracey Siebert,KenPocock,Sally-JeanBell,Ella Robinson,Gordon Elsey, DavidBoehm,Geoff Cowey, Ingrid Oats,PaulSmith,MarkusHerderich, LeighFrancis,PatrikJones,EvelineBartowsky, AlanPollnitz, Elizabeth Waters, KevinPardon, Richard Muhlack,PeterEichinger, MariolaKwitakowski,SakkiePretorius Dillon,JulieMcConnell,DanielCozzolino,DimitraCapone,BobDambergs, Simon Catherine Daniel, Jaro Cynkar, Guzinski,HolgerGockowiak,PaulHenschke,LesJanik,JeanMcIntyre, Wies MariadeSa, Peter Høj,RandellTaylor, MigueldeBarros Lopes,AnthonyHeinrich,DiegoTorrea, AdrianCoulter, Melissa Francis,RaeBlair Mark Gishen,AnnaCatalano,RachelEdwards, AthinaMassis,RhondaPacker, PaulineJorgensen, Hans Muhlack,MerranDaniel,GayleBaldock,GeorgeSkouroumounis, JennyBellon,Lorelie Flood, Jelena Jovanovic,Yoji Hayasaka,SandraLloyd-Davies,HeatherSmyth,KateLattey, KerryWilkinson, Narelle D’Costa,AmandaCook,Heather Donnell Aggie Janusz,JuliaCrossman, Rachel Brown, NicolaSleep,MariaMills,HeatherBrooks, Creina Stockley,