Source: https://apvma.gov.au/node/19446
Timestamp: 2017-05-22 23:20:44
Document Index: 405736717

Matched Legal Cases: ['art 1', 'art 2', 'art 2', 'art 3', 'art 1', 'art 2', 'art 2', 'art 3']

Transcript for Andrew Negri, Australian Institute of Marine Science | Australian Pesticides and Veterinary Medicines Authority
HomeRegistrations and permitsRegistrations and permitsChemical product registrationChemical product registrationApplying for approvals, registrations and variationsApplying for a label approvalWhich products/chemicals require registrationWhich products/chemicals require registrationDoes my veterinary product need to be registered?Does my veterinary product need to be registered?User guide: what is or isn't a veterinary product?User guide: what is or isn't a veterinary product?Animal cosmetic productsAnimal cosmetic productsExcluded animal cosmetic products—acceptable cosmetic messagesAnimal feed productsAnimal feed productsAcceptable nutritional messagesFeed supplements—acceptable nutritional messagesOther requirements for whole unprocessed animal tissueDevices, equipment and physical barrier productsOther products that don't require registrationRegistration self-assessment tool (veterinary)Does my agricultural product need to be registered?Changes to stockfeed and petfood regulationEND product FAQsNovel productsArrangements for products of low regulatory concernArrangements for products of low regulatory concernRegistration modelsRegistration modelsOral electrolyte products for the treatment of horses in exerciseReserved chemical productsListed chemical productExemptionsSubstances not permitted for use on food-producing animals in AustraliaBefore you applyBefore you applyAbout the approval and registration processWho can applyRestricted chemical productsConditions of approval or registration and label approvalPre-application assistanceMinor variationsTechnical assessmentThe risk analysis processTimeframe and feesInterchangeable Constituent Determination - Item 29What to include in your applicationWhat to include in your applicationData guidelinesEvidence of Good Manufacturing Practice (GMP) for veterinary chemical productsEvidence of Good Manufacturing Practice (GMP) for veterinary chemical productsProviding evidence of good manufacturing practice (GMP)Guidance for preparing information listsDossiersRequired informationStatutory criteriaStatutory criteriaActive constituentChemical productChemical productSafety criteriaEfficacy criteriaTrade criteriaLabelling criteriaSatisfying the statutory criteriaAg Labelling CodeVet Labelling CodeGuidance for applicants – submission of international data, standards and assessmentsApply nowGuide to completing an online applicationGuide to completing an online applicationOnline servicesDecision treeIdentification, applicant and nominated agent detailsData and information listsData and information listsUsing the information list editorAccess to informationFormulationManufacturingPackaging and storageLabelsSubmitting an applicationSubmitting third party informationAfter you applyAfter you applyWhat happens nextPreliminary assessmentRequiring samples or further informationRequiring samples or further informationRequiring additional informationRequiring samples to be given for analysisExtended assessment period and maximum response periodRecategorising an application item or module levelRenew a registrationRenew a registrationAbout the application processTimeframe and feesApply nowTechnical policyTechnical policyNo-observed-effect level and no-observed-adverse effect level: use in animal health risk assessmentsAdopted international technical guidance materialAfter registrationAfter registrationChanges to products and activesChanges to products and activesReporting relevant informationAnnual return of activesAnnual return of activesAnnual reporting of actives–frequently asked questionsLevies charged on registered product salesLevies charged on registered product salesDeclaration of leviable valuesNotice of levy assessmentPrimary and secondary applicationsData guidelinesData guidelinesListing of data guidelinesLabelling codesLabelling codesGlobally harmonized system of classification and labellingListing of labelling codesPermitsPermitsBefore you applyBefore you applyAbout the application processTypes of permitsTypes of permitsAgricultural permitsExport, research and miscellaneous permitsMinor use and emergency permitsMinor use and emergency permitsGuide for determining emergency uses or research purposesGuide for determining minor usesPre-application assistanceWho can applyWho can applyFit and proper person testTechnical assessmentTimeframe and feesAbility to comply with conditionsWhat to include in your applicationWhat to include in your applicationData guidelinesDossiersRequired informationStatutory criteriaStatutory criteriaActive constituentChemical productChemical productSafety criteriaEfficacy criteriaTrade criteriaSatisfying the statutory criteriaLabelling requirementsAfter you applyAfter you applyWhat happens nextPreliminary assessmentRequiring samples or further informationRequiring samples or further informationExtended assessment period and maximum response periodRecategorising an application item or module levelApply nowExtend the duration of a permitSearch for a permitManufacturingManufacturingManufacturing licencesManufacturing licencesManufacturers’ Licensing SchemeApplying for a manufacturing licenceWho can applyManufacturing licence feesLicence conditionsAfter you applyVariation of a manufacturing licenceAudits to obtain and vary a licenceApply nowResponsibilities of manufacturersExporting veterinary chemical productsManufacturing overseasManufacturing overseasGMP in overseas premisesRecognition of overseas GMP authoritiesOverseas GMP documentationAcceptable evidence of GMP complianceAPVMA audits of overseas manufacturersContract manufactureContract manufactureRelease for supplyPerforming release for supplyResponsibilities in contract manufactureGood Manufacturing Practice (GMP)Good Manufacturing Practice (GMP)The legislative basis for the APVMA’s role in manufacturing quality and licensingManufacturing PrinciplesVeterinary manufacturing permitAuditsAuditsArranging and undergoing an auditAudit ratings and scoresAudits of Australian veterinary chemical product manufacturing sitesAudits of overseas manufacturing sites by APVMA-authorised auditorsAudit closureThe GMP audit cycleRisk-based scheduling of GMP auditsLicensed manufacturersImport and exportImport and exportImporting agricultural chemicals or veterinary medicinesImporting agricultural chemicals or veterinary medicinesResponsibilities of importersExport certificatesExport certificatesBefore you applyBefore you applyAbout the application processTimeframe and feesWho can applyCommon types of certificatesWhat to include in your applicationWhat to include in your applicationInformation to be provided with a request for a section 70 certificateInformation to be provided with applications for certificates of exportApply nowHolders and nominated agentsHolders and nominated agentsChange a holder or nominated agentChange a holder or nominated agentAbout the application processTimeframe and feesDefinition of roles and actions for holders, nominated and authorised agents and authorising partiesMaking changes to veterinary products and actives—clarification for industryGetting assistance from the APVMAGetting assistance from the APVMAGeneral assistancePre-application assistancePre-application assistanceApplying for pre-application assistanceGetting the most out of pre-application assistancePre-application assistance— timeframes and feesHow to withdraw a request for pre-application assistanceHow to apply for pre-application assistanceTechnical assessmentOther Australian Government requirementsOther Australian Government requirementsDrinking water guidelines—pesticidesLimits on use and disclosure of informationLimits on use and disclosure of informationInformation and the use of informationLimitation periodsApplication informationInformation listsAuthorising partyTrade-related aspects of intellectual property rightsExceptionsSeek review of a decisionSeek review of a decisionReviewing decisionsApproval or variation of an active constituent or registrationPermitsRenewal of registrationCertificates for exporting agricultural and veterinary chemical productChange the holder or nominated agentManufacturing licencesApplications and decisionsApplications and decisionsApplication summariesApplication summariesApplication summariesTransitional arrangementsTransitional arrangementsApplicationsChemical reviews commenced before 1 July 2014Chemicals and ProductsChemicals and ProductsSearch registered chemical products and permitsSearch registered chemical products and permitsSearch PubCRISUsing PubCRISReport a problem with a chemicalReport a problem with a chemicalAdverse Experience Reporting ProgramAdverse Experience Reporting ProgramThe importance of the programScope of the programWho can report an adverse experience?Adverse Experience Reporting Program for veterinary medicinesAdverse Experience Reporting Program for agricultural chemicalsWhat happens nextHow do I report an adverse experience?Using chemicalsUsing chemicalsInformation for farmersInformation for farmersChemicals and farmingUseful links for farmersInformation for veterinariansChoosing a chemicalUnderstanding pesticide chemical labelsConcerns about chemicalsUsing chemicals correctlyUsing chemicals correctlyWorkplace health and safetyProtective clothingSpraying chemicalsUsing veterinary medicinesUsing pool chemicalsChemicals in the newsChemicals in the newsInformation about glyphosate useCurrent review of 2,4-D and IARC’s assessmentHendra virus vaccineHendra virus vaccineProduct registrationSafety, health and side effectsAdverse reactions and how to report themSummary of adverse experience reports made to the APVMA about Hendra virus vaccineRelated linksParkinson's disease and agricultural chemical regulationPyrethrin and pyrethroid use on catsPyrethrin and pyrethroid use on catsAPVMA's roleToxicityGuidance for cat ownersSpray driftPesticides and veterinary residuesChemical reviewChemical reviewListing of chemical reviewsChemicals nominated and prioritised for reconsiderationChemicals nominated and prioritised for reconsiderationSystem to prioritise nominated chemicals for reviewChemical review processChemical review processLegislative basisRegulatory scienceHistory of the chemical review programThe reconsideration processTimeframes for chemical reviewsSupplying chemicals and chemical products in AustraliaSupplying chemicals and chemical products in AustraliaSupply and sale of hormonal growth promotantsHydroponic Product Sellers and agvet chemicalsActive constituentsActive constituentsAPVMA standards for active constituentsActive constituents not requiring evaluationGeneral list of impurities and classes of impurities of toxicological concernProcess for developing a standard for active constituentsApproval of active constituentsTrade-related aspects of intellectual property rights protectionHealth based guidance valuesHealth based guidance valuesAcceptable Daily Intakes for Agricultural and Veterinary ChemicalsAcute Reference Doses for Agricultural and Veterinary ChemicalsFAISD handbookHormonal growth promotantsCompliance and EnforcementCompliance and EnforcementReport suspected non-complianceRecallsCompliance monitoringCompliance monitoringThe ongoing responsibilities of holdersThe ongoing responsibilities of holdersConditions set by the APVMAMaintaining particularsProduct stewardshipCertificates issued under section 70 of the Administration ActEvidential certificatesAdministering holder and nominated agent detailsMonitoring and audit activitiesMonitoring and audit activitiesApproved analystsApproved analystsCriteria for Appointment as Approved AnalystAudits and inspections frequently asked questionsCurrent compliance campaignsGood Manufacturing Practice (GMP)Good Manufacturing Practice (GMP)Arranging and undergoing an auditAuditing of Australian veterinary chemical product manufacturing sitesAudits of overseas manufacturing sites by APVMA-authorised auditorsSwimming Pool Sanitation and the Agricultural and Veterinary Chemicals Code 1994Advertising agricultural and veterinary chemical productsCompliance and Enforcement PolicyCompliance and Enforcement PolicyDisclosure of informationCompliance and Enforcement FrameworkCompliance and Enforcement FrameworkOur compliance and enforcement principlesOur compliance and enforcement authority and powersAPVMA Compliance—our risk-based approachResponsive compliance and enforcementOur powers to manage risk and enforce complianceNoncompliance and recallsPowers to manage risk or enforce complianceEnforcement actionsCivil and criminal actionsAPVMA administration and oversight of compliance and enforcementCompliance and Enforcement guidelinesCompliance and Enforcement guidelinesExemptions from certain offence provisions of the Agvet CodeCancellation or suspension of a permit for noncompliance or prior convictionsCancellation or suspension of a registration or permit to prevent imminent riskConsent to import chemical productsEnforceable directionsEnforceable undertakingsFormal warningsInfringement noticesNotices to attend, give information or produce documents or thingsStop supply or recall noticesSubstantiation noticesSection 99 noticesOur scienceOur scienceOffice of the Chief ScientistAPVMA standard on good regulatory scienceScience FellowsRegulatory Science NetworkThe application of science to regulatory risk assessmentThe application of science to regulatory risk assessmentNanotechnology regulationThe risk analysis frameworkAboutAboutAPVMA basicsAPVMA basicsWhat we regulateWhat we don't regulateWhat we don't regulateProducts we don’t regulateProduct advice and competitionUse of chemicalsWorkplace health and safetyLab testing and product trialsManagement of use of pesticides and veterinary medicinesHow we regulateOur structureOur structureChief Executive OfficerOur boards and committeesOur boards and committeesAdvisory BoardAudit CommitteeRegistration Liaison CommitteeAPVMA cost recovery arrangementsAPVMA cost recovery arrangementsCost recovery questions and answersAcronyms and glossaryAcronyms and glossaryDefinition of termsAbbreviations and acronymsOther informationOther informationCorporate governanceOur functions and powersThe role of the APVMAThe role of the APVMAThe APVMA's regulatory activitiesThe APVMA's regulatory activitiesAccess to safe and effective agricultural and veterinary chemicalsOngoing compliance to ensure safety and efficacyThe Record and RegisterA trusted regulatorA trusted regulatorOrganization of economic cooperation and developmentVICHJoint FAO/WHO Meeting on Pesticide ResiduesJoint FAO/WHO Expert Committee on Food AdditivesHarmonisationCollaborative and overseas assessmentsThe Rotterdam and Stockholm conventionsCommunication, decision making and informationCommunication, decision making and informationCommunication between applicants and the APVMACommunication between applicants and the APVMAWhen we will notify applicantsConsultation and collaborationConsultation and collaborationPublic consultationsTransparencyData handling by the APVMAData handling by the APVMAApproach to managing confidential commercial information (CCI)Applications without written consent for reference productsExternal adviceVision, mission and purposeChemical regulation basicsChemical regulation basicsOverview of agvet chemical regulationThe chemical and permit databasesProduct regulation according to riskLegislative frameworkRisk managementService charterHow we make decisionsHow we make decisionsBackground to the reforms and the section 6A guidelinesOverarching principles and processes for the effective and efficient regulation of agricultural and veterinary chemical productsApprovals and registrationsPreliminary assessmentPermitsLicencesRecategorising applicationsAltering applicationsSection 159 in the context of applications under the Agvet codeLimits on use of informationReconsidering approvals and registrationsVarying relevant particulars and conditionsModule DescriptorsFormulation typesAccess to information held by the APVMAAccess to information held by the APVMAAgricultural and Veterinary Chemicals Code requestsFreedom of InformationFreedom of InformationFreedom of information disclosure logAccess, accountability and reportingAccess, accountability and reportingQuarterly performance statisticsQuarterly performance statisticsQuarterly report: January–March 2017Quarterly report: January–March 2017Analysis by type of applicationMajor and non-technical product applicationsApplication by item numberAverage decision timeOther regulatory functions2016–17 year to date performance overviewQuarterly report: October–December 2016Quarterly report: October–December 2016Analysis by type of applicationMajor and non-technical product applicationsApplication by item numberAverage decision timeOther regulatory functions2016–17 year to date performance overviewGlossary of termsQuarterly report: July–September 2016Quarterly report: July–September 2016Part 1: Overview July–September 2016Part 2: Applications for products, actives and permitsPart 2: Applications for products, actives and permitsPerformance statistics July–September 2016: Applications commencedPerformance statistics July–September 2016: Applications finalisedPerformance statistics July–September 2016: Average decision timePerformance statistics July–September 2016: Performance by assessment typePerformance statistics July–September 2016: Preliminary assessment applicationsPerformance statistics July–September 2016: Staffing considerationsPerformance statistics July–September 2016: Timeframe performancePerformance statistics July–September 2016: Work-in-progressPerformance statistics July–September 2016: Pre-application assistancePart 3: Other timeframe performance results July–September 2016Part 1: OverviewPart 2: Applications for products, actives and permitsPart 2: Applications for products, actives and permitsApplication typesApplications commencedApplications finalisedAverage decision timeAverage decision timeInterpreting average decision timeInterpreting timeframe performancePerformance by type of product assessmentPre-application assistancePreliminary assessmentStaffing considerationsTimeframe performanceWork-in-progressPart 3: Other timeframe performance resultsAPVMA Regulator Performance FrameworkAPVMA legal expenditureAnalysis of time to complete a product applicationAnnual Procurement PlanAuditsInformation Publication SchemeInformation Publication SchemeInformation publication scheme agency planPesticides and Veterinary Medicines Product SalesPublic Interest Disclosure ProceduresAgency contracts over $100,000Indexed list of filesStatutory appointmentsPortfolio Budget StatementsCareersCareersAPVMA Graduate Development ProgramConditions of EmploymentEquity and DiversityProcedures for Determining Breaches of the APS Code of Conduct and for Determining SanctionSocial mediaSocial mediaAPVMA Social Media PolicyAPVMA Twitter policyDisclaimerTerms of UseOnline servicesNews and publicationsNews and publicationsNewsAPVMA relocationAPVMA relocationAPVMA Relocation Advisory Committee terms of referenceRelocation—frequently asked questionsRegulatory updateMedia releasesPublic consultationsPublic consultationsClosedPublic Release SummariesTrade Advice NoticesCrop groupsProjectsProjectsImproved access to chemicalsImproved access to chemicalsThe crop groupings projectThe crop groupings projectCrop groupings project—consolidated responses to phase I consultation—March 2016Lower regulatory approaches to registrationLower regulatory approaches to registrationApplication profiling and registration pathwaysApplication profilingOnline fast-track registration systemExpanded list of notifiable variationsStandards and monographs for low risk productsContestability of the efficacy assessmentContestability of the efficacy assessmentAgricultural scientific reviewersVeterinary scientific reviewersEventsEventsPrevious APVMA eventsPrevious APVMA eventsAPVMA Advisory Board Futures Forum 2014APVMA Advisory Board Futures Forum 2014APVMA Advisory Board Futures Forum 2014 - SummationAPVMA Nanotechnology Regulation Symposium 2014Industry Information and Education Session in Sydney on 1 June 2015Industry Information and Education Sessions in Melbourne on 17 August 2015Program and speaker information—Canberra Industry Information and Education Session 15 and 16 October 2015Industry Information and Education Session in Canberra 15 and 16 October 2015Industry Information and Education Session in Canberra 15 and 16 October 2015Transcript for Andrew Negri, Australian Institute of Marine ScienceTranscript for Chris Lee-SteereTranscript for Distinguished Professor Jim RiviereTranscript for Distinguished Professor Nancy Monteiro-RiviereTranscript for Dr Jason Lutze, APVMATranscript for Dr Phil ReevesTranscript for Dr Raj Bhula, APVMATranscript for Ms Janis Baines, FSANZTranscript for Professor Michael RobertsIndustry Information and Education Sessions in Canberra 8–9 November 2016Program and speaker information—Canberra Industry Information and Education Sessions 8–9 November 2016Industry Information and Education Sessions in Melbourne on 9 May 2016PublicationsPublicationsAPVMA GazetteAnnual reportsCorporate documentsManualsAdverse Experience Reporting Program annual reportsPortfolio Multicultural Plan 2013–15TrainingTrainingTraining workshops and eventsTraining workshops and eventsPermits eLearning module (text only)Registrations eLearning module (text only)eLearning modulesOnline services updateFormsContact usContact usAccessibility This content is current only at the time of printing. This document was printed on 23 May 2017. A current copy is located at https://apvma.gov.au/node/19446 Print this page
You are hereHomeNews and publicationsEventsPrevious APVMA eventsIndustry Information and Education Session in Canberra 15 and 16 October 2015Transcript for Andrew Negri, Australian Institute of Marine Science Transcript for Andrew Negri, Australian Institute of Marine Science
Australian Models for aquatic risk assessment—seagrass study and development of alternative test methods
Thanks everybody for hanging around until this late stage in the day. Thanks to the APVMA for getting me up to talk about the new tests that we're developing for particularly seagrasses and photosystem II herbicides. Just to give you a bit of a background, I'm from Townsville at the Australian Institute of Marine Science. We're a Commonwealth authority. We're publicly funded. We do get some external funding from elsewhere. We have around 200 staff and we basically work around the top end of Australia so everywhere from the Great Barrier Reef right around there to the Ningaloo Reef on the other side. We're essentially a little bit the same as CSIRO.
We've got a range of facilities up there in Townsville. We've got some great lab facilities. We're about to get out on the reef both on the west coast and over on the east coast here. We've got the world's most advanced experimental aquarium system in the world, which was just commissioned about two years ago. We call that the National Sea Simulator.
With today's talk I'm going to be talking about herbicides and the Great Barrier Reef to start off with. Just as an introduction, why and how we became involved as a research agency rather than why do we do it, rather than consultants et cetera. I'll talk then about seagrass and PS2 herbicides and some of the background to the techniques that I'll be discussing later. I'll then get into a couple of acute eco‑toxicology tests that we've developed for seagrass and they are based around the effect on photosynthesis. What I'd like to do then is discuss a longer‑term experiment where we take the effects on the photosystem and then have a look at how that progresses to affect the whole plant. We'll look at a few ways of applying these new acute tests and that will be it.
Herbicides are detected in the Great Barrier Reef. They are applied on farms up and down the coast and if you look here you can see sugar cane in yellow around the Mackay Whitsunday area and up in the wet tropics. We also have a lot of grazing down in around the Fitzroy area and we get a lot of photosystem II herbicides from these coastal zones getting into the marine environment during the wet season when we have these monsoonal rainfalls. I think Chris will be talking a lot about these various scenarios in the next talk.
The photosystem II herbicides are the ones that we're most interested in. The reason for that is that they are the ones that we find in greater concentrations in the Great Barrier Reef. They work by blocking electron transport in photosystem II. They are an excellent pre‑emergent herbicide but the issue with that is that photosystem II is conserved right across pretty much all plants and so they can have some negative effects on non‑target species.
Here is a typical flood plume in the wet season. You can see there's a lot of turbid water heading out there into the Great Barrier Reef. This water contains a lot of particulates, it contains nutrients, and this is where the herbicides get into the near‑shore coastal regions. This is a bit of a close up but when you look from space you can sometimes see flood plumes, which go from literally hundreds to thousands of kilometres—well, the GBR is over 1000 kilometres long and almost the entire length of the GBR can be turbid from these particular flood plumes when you have very heavy rainfalls. You can see here with the turbid water it actually decreases the light that gets down to the seagrass. This is the major threat for seagrass on the Great Barrier Reef. It's actually this reduction in light. The reason we see PS2 herbicides there more than most other pesticides is that they are relatively mobile.
I want to talk today about a few of the key organisms that we're interested in that aren't normally looked at in terms of regulatory testing. Tropical micro‑algae. There's certainly been a lot of eco‑toxicology tests of temperate micro‑algae but is there anything special about tropical micro‑algae? I'm not going to talk much about that today except to say that we are interested in corals, seagrasses, et cetera, but it's the bottom of the food chain that's also very important.
Mangroves. Again, I'm not going to be talking much about mangroves today except that photosystem II herbicides could certainly affect mangroves. They're difficult to work with because they're so large but they are a really key habitat‑forming species in the Great Barrier Reef and in fact all the way around the coast. We shouldn't ignore them.
Corals, which are actually animals but they may be affected by herbicides because they are a symbiotic organism that host micro‑algae within their cells, they host dinoflagellates and these provide about 70 to 90% of the energy that the coral needs for growth and reproduction. If there's an effect of the herbicides on the dinoflagellates this may have a flow on effect to the coral itself. Finally seagrass, which is another very important species. It creates a very important habitat and it's an important food for various organisms on the reef.
Why did we become interested? It was about in the year 2000 when photosystem II herbicides were first detected in the Great Barrier Reef Marine Park. Around the same time there was a new instrument called the pulse amplitude modulated fluorometer which held great promise in being able to detect in a non‑invasive way effects of various contaminants or other pressures on plants. The Reef CRC at the time asked us whether or not this could be a useful technique to look at the effects of photosystem II herbicides that had just been discovered on corals. Here you can see a diver with a diving PAM, it's an underwater pulse amplitude modulated fluorometer. You can put it up against a plant or a coral and you can measure a proxy for the efficiency of photosynthesis in that plant or in the coral.
I'm just going to give you a very brief background on how it works. When light hits photosystem II in a plant, the energy can be either used for photochemistry or it can be emitted as fluorescence or dissipated as heat. The PAM fluorometer measures this fluorescence in two different ways. You use this ratio to describe the photosynthetic efficiency. It has a very good correlation in most scenarios with photochemistry.
When PSII herbicides get into the system they block this electron transport in photosystem II. Once this happens, if there's relatively high light, you get oxygen radicals formed within photosystem II and this causes damage. The other thing is that you get an increase in fluorescence, and this changes the ratio, it drops the ratio and indicates a drop in photosynthetic efficiency. You can measure the amount of binding of the herbicide to photosystem II, and you can measure damage to photosystem II using different measurement techniques with the PAM fluorometer.
The advantage of the PAM fluorometer is that it is non‑destructive and it's rapid. It has very high precision. It doesn't matter how many chloroplasts you've got there, it's the ratio of the fluorescence that's important so it's very precise. Importantly it measures the effects of PSII herbicides at the site of action in the plant.
I've shown you there a probe but you can also use it in a weld plate format. The imaging PAM takes, a two‑dimensional picture across surfaces as well. The disadvantage is that it's not as sensitive to other contaminants that don't have direct effect on photosystem II. However, if a contaminant affects photosystem II and damages photosystem II then it may be applicable in that case. It's used in heat stress studies in corals a lot before they bleach. Finally and we'll be talking about this, when you use a technique like this you need some supporting evidence to indicate whether this technique has any ecological relevance or not.
The initial focus of the work back in about 2003, we exposed corals to herbicides and we saw certainly there was reduced photosynthesis, there was damage to photosystem II as indicated by the PAM and we saw coral bleaching. You can see that this coral over here, although it's alive, the coral has actually ejected the symbiotic micro‑algae to reduce the amount of photo‑oxidated stress in the coral itself. It can live for a little while like that but if it doesn't get a population of symbiotes back it will run out of energy. The other thing that we found and particularly Ross Jones's work back then who works with us now was that diuron was certainly one of the most potent of the herbicides that were detected in the Great Barrier Reef.
It was around this time that the APVMA were doing their preliminary review on Diuron and we were working on our corals. We hadn't published anything at that point. We got a visit from DuPont at that time. It was really interesting because they knew a hell of a lot more about Diuron than we ever would and still do. They had several eco‑toxicologists that came along and there was some really good information transfer there. They had two questions, why the focus on Diuron and AIMS is a federal government agency and we see ourselves as the honest broker so we certainly weren't picking on Diuron. It was just that Diuron was detected in the marine environment much more often than other herbicides and its high potency made it really the pesticide that we really wanted to use as a reference. That one was easy to answer.
The next question was, are the effects of PAM fluorometry at all environmentally relevant? That was a tough question. What we did after that was put a couple of PhD students onto this problem. The first one was Marie Magnusson and she did a lot of work in a range of areas. One of the things that she looked at was whether or not there was any correlation between traditional micro‑algal growth assays and biomass assays over 72 hours with the results you get from PAM fluorometry. Here you can see a series of flasks using a typical regulatory‑type test. They've been exposed to different concentrations of herbicides that increase as you head from left to right.
This is part of Marie's results. She used micro‑algae that were isolated from the Great Barrier Reef. You've got the inhibition of growth on this axis and on this axis you've got the inhibition of photosynthetic efficiency from the PAM fluorometer. You can see that there's a really nice correlation. The PAM fluorometry data is much tighter than the growth data but you've essentially got a one‑to‑one correlation over that 72‑hour period. The reason for this is that when the herbicide affects the photosynthesis, there's very little energy storage in the diatoms and in the green algae that she looked at and there's a pretty much immediate effect on growth, which isn't the case with corals and seagrass. They're much more complicated. We would consider that in many situations PAM fluorometry can be a very good and ecologically relevant indicator for eco‑toxicology for micro‑algae at least.
More recently there's been a consensus that if any organisms are at risk from herbicides, it's much more likely to be seagrass than corals. We've moved into that area a bit more in recent times. To recap, seagrass are habitat‑forming. They are food for dugongs and turtles and I mentioned before that they are under threat from light limitation from those flood plumes I showed earlier on. When you have flood plumes, and the turbidity can be increased for very long periods of time, for up to months and recent data indicates for up to six months, you get an attenuation of light. Over years we have very heavy rainfall in the Great Barrier Reef because these sediments keep re‑suspending over time and eventually they flush out of the system. This is when you find dead turtles and dead dugongs up on the beach, after this period of time. Seagrass are really important for the people in the north that are interested in them.
There was some very early work that had been done on exposing diuron or exposing seagrasses to diuron, work by Haynes, and they showed that there was an effect on photosynthesis of 0.1µg per litre which is pretty low. It's lower than concentrations that have been found in the marine park which is up to around 1µg per litre in near‑shore environments. But the work wasn't that well designed to answer an eco‑toxicological problem. They didn't do concentration response curves, for instance. We needed to do something a little bit more robust.
In about 2011 the National Environmental Research Program (NERP), we were lucky enough to get some funding in that to do some work on pesticides. We did some work on persistence. We also did some work on toxicology. Two of the questions were to figure out how toxic were a range of pesticides to a range of seagrass species using PAM fluorometry. The other one was, as I mentioned before, if there's an effect on photosynthesis how does this translate to whole plant effects in the long term?
I'll just start with one of our acute tests. Here's some seagrasses that are potted. We bring them in from the environment. We acclimate them in the lab. They've got their root rhizome complex there so everything's intact. We looked at two species and four herbicides. We did a 72‑hour flow through experiment. I'll explain why in a minute. We measured the inhibition of photosynthesis and damage to photosystem II. We also measured growth but there was no impact on growth over that period. That's just an image of what the lab looked like before the Sea SIM was developed. It was still a pretty good system.
Here are some of the results from that initial study. Here you can see the inhibition of photosynthesis is measured by the PAM fluorometer and as the herbicide concentration increases you get an increase, you get inhibition of photosynthesis and diuron is quite a lot more potent than tebuthiuron, for instance. From concentration response curves like this we can drive the IC50 values, that's the concentrate, the inhibition concentration. That can then be used to define how potent one toxin is against another or how sensitive one organism is against another. We can also derive IC10s for that which we would often consider as the threshold for toxicity.
We certainly found effects of diuron at concentrations that can be found in the Great Barrier Reef. I think one of the important things is that we found that as you add diuron to the seagrass the effect happens relatively quickly and by 24 hours it's reached a maximum. It doesn't matter if you run that experiment for seven days. You still get a similar inhibition of photosynthesis. These are results from 72 hours but they are identical to the results we had at 24 hours and identical to some of the results we had over an 11‑week period in terms of inhibition of photosynthesis. It means we can probably get away with shorter assays.
This is just an example of one of the data sets from that paper we published a couple of years ago. Two different seagrass species. The IC50s, the IC10s are pretty close to each other. It's a very precise method. Tebuthiuron up at the other end of the scale, atrazine somewhere in the middle.
The next thing we did was to see whether or not we could develop a miniature test. In this test we used a different species which had leaves which were about a centimetre long. We wondered because we do this with corals. We don't put entire corals into an assay. We actually snip off branches of corals, we heal them and we put them in assays, so we wondered if we could do the same with seagrass. We got another student on board, Adam Wilkinson. He would snip these leaves off and put them into a 12‑well plate to see if he could develop an assay that was as valid as the previous one that I described with the potted plants. We wanted to see whether the response and sensitivity was similar and whether or not we got those results in a relatively high throughput compared to these big pots in a shorter duration.
This is a series of images of two leaves starting at zero hours. This leaf is in controlled conditions and you can see by the blue colour that it's got a high level of photosynthetic efficiency over that 24 hours. In fact, we can run it for longer. We can run it for 48 hours and the photosystem is still intact over that time. When we add 10µg per litre of diuron just to give a dramatic effect, you can see that it takes up the herbicide and it increases the fluorescence as indicated by the colour changing from green through the dirty red on the right‑hand side. You can see that the maximum effect is happening somewhere between eight and 12 hours.
One of the helpful things is that as opposed to weeds on a farm which often take up the herbicide through the root system, the seagrass and many other aquatic plants take up many of these herbicides actually straight through the leaves. It was great they didn't take it up through the vascular system where we would snip them there so it looked to be a pretty good system.
This is just a graph of the potted plants that I mentioned before, of the species we used, of the same species we used in the weld plates versus the weld plate data for inhibition as we increased the concentration of diuron. We also used hydroponic set ups. It was pretty simple though. We just took the seagrass, we removed the soil and sediments from the roots, we let them acclimate for a few days and we put them into the same system. They all had almost identical responses to diuron using the PAM fluorometer. The IC50s that we got from that were very similar to what we got from the other species, the potted plants. It's a very robust system. It's idea for comparing one plant against each other or herbicides against each other.
What we wanted to do next was to try and link the effects on photosynthesis with the effects on the whole plant and to do that we needed to run the experiments for longer. Again, we used two species. We used diuron as the reference, 11 weeks and a flow‑through system. We looked at the inhibition of photosynthesis and damage to photosystem II, but in this case we looked at the effects on the energetics of the plant which is where we expected the other sub‑lethal effects to be and also the growth and survival over that 11‑week period.
I'm just going to show you very much a summary of the results. Or the numbers that you see are the effect sizes when we had a significant result. If there was no number there was no significance or we didn't show it. I just want to try and keep things simple here. You can see that we've got some different concentrations of diuron here. We've got effects on the photosynthesis. Significant effects at 0.3µg per litre which increases when we increased the herbicide. The same goes with damage to photosystem II although it was not quite as sensitive.
If we look at the CN rations and delta‑13 in the leaves, when these increase it indicates that there is less carbon fixation through photosynthesis over that period of time. You can see decreases there as a bit of an indicator that there's less carbon from photosynthesis. What's really important is the starch though. The starch is how the plants store their energy. They store it in the root rhizomes which are really big on seagrasses. You can see here that over that period the energy level or the energy had dropped right down. Even though we didn't see an effect on growth and mortality until the highest concentration, that those seagrasses weren't in good nick at that particular time.
Here is just a diagram of seagrass and you can see that there's a lot of space here for storage of starch in that root rhizome scenario. It's a lot different to something like duck weed which is often used in aquatic toxicology. It's very difficult actually to use the shorter assays for a week or two weeks and expect to see changes in growth in these leaves because they just keep pumping the energy into the leaves to let them grow more. If you use very high concentrations of herbicides which are unrealistic you probably wouldn't get that effect, they would probably just die straight away but we wanted to keep things relatively environmentally sound.
Also these different leaves all grow at different rates. These ones are not growing at all. These ones are growing superfast and these guys are growing somewhere in the middle so it's very difficult to get accurate growth measurements. What that allowed us to do was come up with a conceptual model for the effects of PSII herbicides on seagrass. Started off with a sustained lowering of the photosynthetic efficiency and damage to photosystem II. This led to less carbon fixation. The energy in the root rhizomes was then mobilised into the leaves so the leaves could capture more light and the stored energy was then consumed and at that point you finally got mortality and effects on growth rates in the leaves.
Interesting thing about this is that's a pretty good news story for seagrass in some respects but in other respects these effects are almost identical to the effects you see when you have light attenuation, from sediments in the water, from those turbid events when you have the river water flowing into the marine park. It could be, and we suspect that if there are any effects of herbicides on seagrass in the Great Barrier Reef, it's most likely that they're just adding to the effect in a very similar way to the light attenuation because their modes of action are very similar from the effects on photosynthesis.
I'm just going to quickly show you some of the applications for some of the new tests. For the miniature test that I explained, it's great because you can just apply it so quickly to a range of different scenarios. In this case Adam exposed the seagrass to different temperatures at the same time as different diuron concentrations. You've got high inhibition of photosynthesis up here and you can see that there's this surface of effect and you actually get a great effect of the herbicide at the extreme temperatures. We've seen exactly the same thing with corals but we haven't been able to do this many treatments all at once because this is about 36 different treatments all combined together.
The other thing that you can do is to do matched data sets. Under identical conditions we looked at 10 different photosystem II herbicides and you can look at their IC50s and you can rank them in terms of potency against seagrass. We also looked at an emerging contaminant, an emerging PSII herbicide as well.
I think this is a really important one. It is about ground‑truthing some of the guidelines that were developed for water quality. This is a species sensitivity distribution for diuron that's being developed or proposed for the new ANZECC guidelines. You can see that as you increase the herbicide more species become affected as you go up that curve. Down the bottom here you can see that if your concentration of herbicide is down here, you would expect 95% of species to be protected. The ANZECC guidelines or the new guidelines, the proposed guidelines, might have a 85% species protection set at 0.3µg per litre for diuron. In fact, in the marine park where it is high conservation value world heritage area, I expect that the Marine Park Authority will apply the 99% protection level which is 0.08µg per litre which is quite a lot lower than what it is at the moment.
If we just compare where our effects on photosynthesis from the PAM fluorometry fit here, we're seeing a 10% effect at about 0.3µg per litre, and we're seeing a 50% effect up here on the curve as well. I'd be quite happy to say that you'd get a reasonable amount of protection of marine organisms down here and that seagrass would be well protected down there as well.
I think days like today are fantastic where researchers get to meet end users. Up in the north we've been holding a pesticide working group for the Great Barrier Reef because it's quite an issue up there. It's considered one of the main threats to the Great Barrier Reef in terms of pollution after nutrients and sediments. We've been holding working groups up there as part of the NERP project up until 2014 and we hope to continue something like that into the future as the new research program NESP comes on board.
I'd just like to finish up by making some conclusions about PAM fluorometry. Remember it measures PSII herbicide effects at the site of action, which is handy for that group of pesticides, herbicides. It reaches a maximum after two hours for micro‑algae, up to four hours for some herbicides with some seagrass species. We have to test that and make sure that we're at the maximum. But you don't really need to do experiments for longer if effects on photosynthesis are your end point. Measures damage to photosystem II. The effects on micro‑algae as measured by the PAM fluorometer can be ecologically relevant but it's a much more complicated scenario if you're talking about corals and seagrass but you can see effects on reproduction in corals and you can see effects on the health of the seagrass and on their growth and survival.
Just some general conclusions now. I think that it's important, the reason we get involved in this is we think it's important that regulations and guidelines take into account the organisms that add value to world heritage areas like the Great Barrier Reef, these species that we really want to protect like corals and seagrasses, et cetera. The effects of PSII herbicides on seagrass are probably not going to effect seagrass by themselves in the marine environment. They probably add to the pressures so when we do laboratory tests the results we get really probably underestimate what happens in the real world because there are other effects of low salinity, et cetera. Maybe the formulations increasing toxicity as well. It hasn't been tested. I think that some of these, if we have new photosystem II herbicides then techniques like this are ideal for measuring their relative potency. We need to continue to communicate because by communication we can tailor the research that we do to better meet the needs of the end users.
I'd just like to thank everybody for turning up today but also our research team, our collaborators at James Cook University and UQ, NERP who funded much of the work and also the APVMA. Thanks very much for flying me down for this. Thank you.
Errors and omissions excepted; check against delivery.URL: https://apvma.gov.au/node/19446Content last updated: 11 December 2015Content last reviewed: 11 December 2015
EventsPrevious APVMA eventsAPVMA Advisory Board Futures Forum 2014
Industry Information and Education Session in Canberra 15 and 16 October 2015Transcript for Andrew Negri, Australian Institute of Marine Science