CONFERENCE UPDATE: WHY YOU SHOULD BE THERE
JUNE 2017 | VOLUME 25 No. 5
Myrtle rust arrives in NZ Karin Kos Lacebark: a star performer Linda Newstrom-Lloyd and Angus McPherson How do labs conduct the MPI tests? Analytica Laboratories South Canterbury field day Rebecca Lees Visit to Bayer Bee Care Don MacLeod
helping take your bee products TO THE WORLD!
I would like your vote for the upcoming Apiculture NZ Board elections.
Our united industry has potential to bring more export revenue into New Zealand. This will happen when we have an industry body with the determination, resource and skill to: 1. TELL THE NZ SOURCE STORY: We have a natural advantage - our location, native flora, culture and heritage. We must develop and promote this shared story, just like other successful primary sectors. 2. SUPPORT CONSUMER TRUST: We need a recognised industry endorsement of world-class products that meet robust standards for safety and authenticity. This will put product from New Zealand ahead of international competition. 3. AGREE ON ONE MANUKA GRADING SYSTEM: Deal with the confusion in the marketplace; one industry grading system for Manuka honey. I understand the value chain from hive to consumer and have the experience necessary to enable our industry to grow and thrive.
VOTE FOR TONYWRIGHT. STANDING FOR A POSITION IN THE MARKET SECTOR tony.wright@comvita.com 021 386 710
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TABLE OF CONTENTS
Star performers part 4: Lacebarks for autumn pollen and nectar Apiculture health and safety programme formed Wanganui Beekeepers’ Club successfully appeals abatement notice Bee educated at Limehills School Wide-ranging discussions at South Canterbury field day
Myrtle rust arrives in New Zealand
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Conference update
Chemical changes in maturing manuka honey How do labs conduct the MPI manuka tests? Visit to Bayer Bee Care: Monheim am Rhein, Germany Obituary: Terence Charles Gavin (Terry) Manuka Health unveils $3.5 million Wairarapa Agriculture Centre
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From the colonies
Keep your hives disease free
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Front cover: Myrtle rust (Austropuccinia psidii) on myrtus communis (common myrtle). Photo: Forest and Kim Starr, Starr Environmental. https://www.flickr.com/photos/starr-environmental/24278125284/sizes/l This work is licensed under a Creative Commons Attribution 2.0 Generic License. Some rights reserved by Starr Environmental. Photograph orientation changed from landscape to portrait. Link shared courtesy of the Ministry for Primary Industries.
EDITORIAL/PUBLICATION (excluding advertising): Nancy Fithian 8A Awa Road, Miramar, Wellington 6022 Mobile: 027 238 2915 Fax: 04 380 7197 Email: editor@apinz.org.nz ADVERTISING INQUIRIES: Certa Solutions, PO Box 2494, Dunedin 9044. Phone: 0800 404 515 Email: beekeeper@certasolutions.nz PUBLICATIONS FOCUS GROUP: Frank Lindsay 26 Cunliffe Street, Johnsonville, Email: lindsays.apiaries@clear.net.nz DEADLINES FOR ADVERTISING AND ARTICLES: Due on the 6th of the month prior to publication. All articles/letters/photos to be with the Editor via fax, email or post to Nancy Fithian (see details above). Articles published in The New Zealand BeeKeeper are subject to scrutiny by the Apiculture New Zealand Management Team. The content of articles does not Wellington 6037 Ph: 04 478 3367
The New Zealand BeeKeeper is the official journal of Apiculture New Zealand (Inc.). ISSN 0110-6325 ISSN 2537-8058 (Online) Printed by Certa Solutions, PO Box 2494, Dunedin 9013, New Zealand ApiNZ website: www.apinz.org.nz
necessarily reflect the views of Apiculture New Zealand. © The New Zealand BeeKeeper is copyright and may not be reproduced in whole or in part without the written permission of the Publisher, Apiculture New Zealand (Inc.). CONTACTS TO THE NEW ZEALAND BEEKEEPING INDUSTRY: Rex Baynes, AFB PMP Manager PO Box 44282, Lower Hutt 5040 Email: rbaynes@ihug.co.nz Ph: 04 566 0773 American Foulbrood Management Plan www.afb.org.nz
MANAGEMENT TEAM: Chief Executive Officer Karin Kos Email: ceo@apinz.org.nz Secretary Email: info@apinz.org.nz Accounts and Subscriptions Pauline Downie Email: memberships@apinz.org.nz PO Box 25207, Featherston Street,
AsureQuality Limited Phone: 0508 00 11 22 www.asurequality.com EXOTIC DISEASE AND PEST EMERGENCY HOTLINE 0800 80 99 66 www.biosecurity.govt.nz
Wellington 6146 Ph: 04 471 6254 APICULTURE NZ BOARD REPRESENTATIVES: Dennis Crowley
Barry Foster Stuart Fraser Sean Goodwin John Hartnell Ricki Leahy
Pollinator Incident Reporting Form: http://www.epa.govt.nz/Publications/ Pollinator_incident_reporting_form_2014. docx
Peter Luxton Russell Marsh Paul Martin Bruce Wills (Chair)
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NEW ZEALAND BEEKEEPER, JUNE 2017
MYRTLE RUST ARRIVES IN NEW ZEALAND CHIEF EXECUTIVE’S REPORT Karin Kos, Apiculture New Zealand Chief Executive The arrival of myrtle rust to New Zealand shores last month was disappointing but unfortunately not unexpected, given its recent spread along the east coast of Australia and the fact that it is carried by the wind and on people, vehicles and equipment.
It’s a particularly nasty fungus that attacks various species of plant in the myrtle family including pōhutukawa, mānuka and other natives, and also feijoa and guava trees. But at this stage we don’t know how myrtle rust will behave in New Zealand conditions and the impact it might have on our native plant species like mānuka. While bees can transfer the myrtle rust spores from plant to plant, our best information tells us that they do not transfer the spores into the hive. Tests in Australia (where myrtle rust is widespread) did not find myrtle rust spores in beehives. The Aussie experience We’ve talked to our counterparts in Australia, including the Australian Honey Bee Industry Council, as well as individuals involved in the industry. They say that when myrtle rust arrived in Australia in 2010 there were concerns about the impact it might have on honey production; however, Australian beekeepers are still producing jelly bush honey (a Leptospermum species) in affected areas. The key lessons learnt in their response to this disease is the need for a coordinated government, industry and stakeholder response; collaborative research working with countries already impacted by the disease; and longer-term initiatives like seed banking and resistance breeding. These initiatives make a lot of sense and it’s good to see that Kiwi research organisations have already been working with experts in the USA (including Hawaii), South Africa, Australia, Colombia, and Brazil to gain an understanding of how the fungus could affect native and exotic trees in our country. New Zealand researchers also met in Christchurch last month to start to look at what research we currently have and what will be needed over the longer term. For our
Myrtle rust (Austropuccinia psidii) on Agonis. Myrtle rust is a serious fungal disease that affects plants in the myrtle family (family Myrtaceae). Photo courtesy of the Ministry for Primary Industries.
industry, practical measures like seed banking and resistance breeding programmes for New Zealand species are critical. Here at home So what about local beekeepers and the impact on our industry? At this stage MPI has been appealing to beekeepers in affected areas (at time of publication Kerikeri and Waitara/New Plymouth) to not move their beehives if possible, while they gather further information. Fortunately, most hives have been wintered down for the season, so very little movement is expected anyway. MPI also
is advising that there’s no need for any drastic action such as destruction of any beehives. To help with the knowledge on the ground and key contact points, Apiculture New Zealand has provided MPI with local beekeeper and industry liaison people (Paul Martin, ApiNZ Board member and Rob Murray of Tai Tokerau Honey) in Whangarei and Toby Annabell of Egmont Honey in Taranaki. In addition, Apiculture New Zealand will continue to provide regular updates to its members, and is working with MPI to develop a set of management guidelines on myrtle rust for beekeepers.
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Next steps Apiculture New Zealand meets regularly with the wider Government-led Governance Group, which directs the immediate response (containment) to the outbreak and considers what will need to be done if myrtle rust cannot be contained. This includes developing a longer-term community and industry engagement programme, effective research programmes, and information gathering and analysis to understand the economic impact of myrtle rust and how it could potentially affect our economy, including our valuable honey industry. As these take shape, we will be in a position to provide more information to industry, including ongoing responses and managing the disease from a beekeeper perspective. Myrtle rust will also be a topic of discussion at the upcoming ApiNZ National Conference, with speakers from MPI and from Australia. IF YOU BELIEVE YOU HAVE SEEN SIGNS OF MYRTLE RUST: • do not touch it or the plant • take a photo of the rust and the plant • call MPI’s Exotic Pest and Disease Hotline on 0800 80 99 66. If you have any questions about myrtle rust, please email info@mpi.govt.nz. For full details about the fungus, go to https://www.mpi.govt. nz/protection-and-response/ responding/alerts/myrtle-rust
Myrtle rust on Agonis. Photo courtesy of the Ministry for Primary Industries.
Close-up of myrtle rust on pohutukawa. Yellow bumps and brown patches are typical of myrtle rust. Photo courtesy of the Ministry for Primary Industries.
SEE YOU AT CONFERENCE I’m looking forward to attending our annual national conference in Rotorua from Sunday 9 July to Tuesday 11 July. I’ve been meeting with the steering committee recently and this year’s conference is shaping up to be the must-attend event of the year. The conference will be a great opportunity for everyone to get together, hear from inspirational and informative speakers, and discuss some of the big-picture issues and opportunities facing our sector. It is also a showcase for our industry, we’re expecting over 1000 attendees and the largest trade exhibition to be assembled at an apiculture industry conference. Don’t miss out and book your place at conference today. To find out more, go to http://apicultureconference2017.co.nz
NATIONAL CONFERENCE 2017 Rotorua, Sunday 9th, Monday 10th, Tuesday 11 July 2017
Call for entries...
AWARDS & COMPETITIONS
The Roy Paterson Trophy An award for the best innovative idea, invention or new technology to benefit New Zealand beekeepers, whether in the field or at the factory. Entries close – 8.00 am Sunday 9th July 2017. ApiNZ 100% Pure New Zealand Honey - National Honey Competition Where quality, innovation, and pride take centre stage. Entries close for the main competition 2.00pm Sunday 9th July 2017. For the Airborne Commercial Class – Wednesday 31st May 2017. ApiNZ Ecrotek - National Photo Competition A great opportunity to showcase your skills as a photographer, from that classic snap to the proverbial blooper. Entries close 12.00 noon Sunday 9th July 2017. ApiNZ Peter Molan Award - Excellence in Apiculture Science Recognition of an outstanding contribution in the field of apiculture science, this is a special award and comes with a substantial Science Grant. Nominations close on Sunday 30th June 2017. ApiNZ Unsung Hero Award - The Buzziest Bee Recognises the efforts of a member of the apiculture industry who has gone that extra mile, who has given his or her time for the betterment of industry without seeking recognition or reward. Nominations close on Sunday 30th June 2017. For full details on how to enter or nominate an award recipient please go to: www.apicultureconference2017.co.nz Thank you - Conference Steering Committee 2017
Conference website: www.apicultureconference2017.co.nz Conference hotline: 09-520 9198
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NATIONAL CONFERENCE 2017 BEE-ING PREPARED—BUSINESS : KNOWLEDGE : RISK Sunday, Monday, Tuesday 9–11 July ENERGY EVENTS CENTRE – ROTORUA
WHY SHOULD YOU BE THERE? • MPI Mānuka definition – the financial impact
• GREX – export requirements, market access, certification • Myrtle Rust – what could this mean to your business? • Bee Health – pesticides and neonicotinoids • Varroa Management – an international perspective • Global market trends and challenges • Health & Safety – the must do’s • Financial risk management and your business
• Ethics and best practice outcomes • Science – key research updates • Extensive Trade Exhibition
The draft programme details are available now on the conference website: www.apicultureconference2017.co.nz Keynote speakers include Randy Oliver (USA), Yves Le Conte (France), SimonWilliams (Australia), with science input from Australia on myrtle rust. From New Zealand: Dr Mark Goodwin and Dr David Pattemore (Plant & Food), Bryan Wilson and Rachel Linwood (MPI), Dr Pike Brown (Landcare Research), Dr John Mackay (dnature), Richard Toft (Entecol – Wasp Control), Dr Catherine Beard and Arna Litchfield (DOC), Marieke Soeter (EPA), Dr Rebecca Martin (MPI – Myrtle Rust), Iain McLeod (BNZ), Mary-Anne Thomason (Kintail Honey), Prof Peter Dearden (University of Otago), Jason Ward (H&S), Tony Wright (Standards & Compliance).
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CONFERENCE 2018 Confirmed for Blenheim, 1–3 July, Marlborough Convention Centre.
Conference begins for all delegates on Sunday 9 July. We encourage you to register early, book your accommodation and nominate your workshop preference as soon as possible, as limits may apply. Charity auction recipient – “St John” – please support this worthy organisation. Details of all the Competitions and Award opportunities and how to nominate worthy recipients are available on the conference website.
Delegate and trade exhibition numbers will be restricted due to venue capacity, so please register your interest early. Conference 2018 website: wwwapicultureconference2018.co.nz Sincere regards, Conference Steering Committee 2017
• The Roy Paterson Trophy • ApiNZ 100% Pure New Zealand Honey – National Honey Show • ApiNZ Ecrotek – National Photographic Competition • ApiNZ Peter Molan Award – Excellence in Apiculture Science • ApiNZ Unsung Hero Award – “The Buzziest Bee” Please do not hesitate to nominate someone if you believe they meet the criteria. Registration for conference is open. You can register online or by phoning the conference team, who will mail out a hard copy or provide assistance where required. Conference 2017 website: www.apicultureconference2017.co.nz Conference hotline: 09 520 9198
NOTICE OF THE 2017 AGM of Apiculture New Zealand to be held at the Sir Howard Morrison Performing Arts Centre in Rotorua
The AGM will commence at 10.00 am Wednesday 12 July 2017
Members are invited to attend the AGM for Apiculture New Zealand, which will be an opportunity for all members to hear about the Association’s progress over its inaugural year, and plans for the future.
Karin Kos Chief Executive Officer
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CHEMICAL CHANGES IN MATURING MANUKA HONEY RESEARCH Merilyn Manley-Harris, School of Science, University of Waikato The University of Waikato honey chemistry research group has recently published a fourth journal article in their series describing chemical changes in maturing mānuka honey; this article describes the development of 5-hydroxymethylfurfural (HMF) during maturation (Grainger, Owens, Manley-Harris, Lane, & Field, 2017).
Queen bee in a hive in Nelson. Photo: Elliott Kennedy.
Effect of temperature Probably the most important take-home message from this paper is the effect of temperature. At 4 o C, HMF remained at a very low level for 300 days; at 27 o C, the 40 mg/kg limit was reached at ~230 days; at 32 o C, the limit was reached at ~190 days and at 37 o C the same limit was reached at only ~90 days. Thus warming honey, in order to accelerate conversion of DHA to MG, can have the unwanted side effect of increasing the HMF. Warming can also reduce the final levels of MG, as shown in Parts I–III of this series. Research at Waikato and overseas indicate that different honeys can respond differently at the same temperature. This is undoubtedly due to the levels of components other than sugar for example amino acids, phenolic acids and water, which can have the effect both of accelerating production of MG or HMF but also consuming the product. Reference Grainger, M. N. C., Owens, A., Manley-Harris, M., Lane, J. R., & Field, R. J. (2017). Kinetics of conversion of dihydroxyacetone to methylglyoxal in New Zealand mānuka honey: Part IV – Formation of HMF. Food Chemistry, 232, 648–655. http://dx.doi. org/10.1016/j.foodchem.2017.04.066 [Editor’s note: a summary of the first three papers in the series was published in the May 2016 journal, pages 24–25.]
Because of its perceived toxicity, the international Codex Alimentarius , with certain exceptions, indicates a limit of 40 mg/kg of HMF in honeys and, although there is no legal domestic New Zealand limit, exporters usually try to ensure their product is below 40 mg/kg. HMF forms by a chemical reaction in maturing honey of any type: it is principally formed from fructose, which constitutes roughly half of the sugars present in the honey. Because it is a chemical reaction, it will continue for the entire life of the honey, processing, packaging, retail and domestic use. In mānuka honey the two reactions that form MG and HMF are unrelated but can impinge upon each other indirectly, because both reactions depend upon the presence of proton donor and acceptor molecules (amino acids, phenolic acids, etc.), and DHA, MG and HMF can all consume amino acids by reacting with them. Water, which can act as either a proton donor or acceptor, is very limited in honey and tied up by the sugar molecules. The research found that the increase of HMF versus time was first-order (logarithmic) during an induction period and zero-order (linear) thereafter; the duration of the induction period depended upon the temperature and was shorter at higher temperatures. A relatively simple computer model was able to simulate the formation of HMF in mānuka honeys at 37 o C and the behaviour of mānuka honey was mimicked by clover honey doped with DHA, or by artificial honey with DHA and appropriate amino acids added.
WE WANT YOUR PHOTOS! The Publications Committee welcomes photos for the journal. Pop a camera in the truck and snap away when you find something interesting. The safest way to supply a digital file is in a high-quality jpeg format. If you’re thinking big (such as a potential front cover photo), these need to be as large as possible (300 dots per square inch (dpi) at the size they are to be used, in portrait format (vertical rather than horizontal). Regular digital photos are only 72 dpi, so are not suitable for the front cover. Please provide a caption and the name of the photographer so we can credit them. Email photos and captions to editor@apinz.org.nz
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NEW ZEALAND BEEKEEPER, JUNE 2017
HOW DO LABS CONDUCT THE MPI MANUKA TESTS? RESEARCH Steve Howse, Dr Gary Depree, and Dr Maria Tourna Analytica Laboratories In April 2017, MPI announced a proposed new definition for mānuka honey, which includes some new requirements for laboratory testing. This article gives an overview of how the testing is carried out by laboratories, and some thoughts about how to understand the results. The testing uses well-established techniques, and prices are in the range of $180–$200 + GST per sample if all five tests are requested for a honey sample.
MPI’s proposed mānuka definition was announced on 11 April 2017 A great deal of useful information is available on MPI’s website (www.mpi.govt.nz) about the proposed definition of mānuka honey. The proposed definition includes testing for a combination of five attributes (four chemicals and one DNA marker from mānuka pollen) to distinguish mānuka honey from other honey types, and to identify monofloral and multifloral mānuka honey. A summary of these tests and required results is given in Table 1. All four chemical markers can be analysed in a single test carried out by the laboratory. The DNA pollen test is a separate test using entirely different equipment and testing procedures. Take care if your test results are close to the required levels If a honey sample is tested a number of times, you will not get exactly the same result each time. Because there is variability in both a sample and in the testing process, results will naturally vary across a range. Laboratories measure this when setting up a test, and refer to it as Uncertainty of Measurement (UoM). Many readers will have experienced this when asking for re-tests of samples in the past. UoM is very important to take into account when comparing your test results against required levels such as those in the proposed MPI mānuka honey definition. If a result for a sample is quite close to the threshold, there is a chance it will produce a result on the other side of the threshold if re-tested (by you or by someone else). For example: • Assume that a method has a UoM of plus or minus 10%, and you are aiming for a minimum of 400 mg/kg.
Required levels for multifloral mānuka honey
Required levels for monofloral mānuka honey
Chemical markers 3-phenyllactic acid
Greater than or equal to 20 mg/ kg and less than 400 mg/kg
Greater than or equal to 400 mg/kg
2’-methoxyacetophenone 2-methoxybenzoic acid 4-hydroxyphenyllactic acid
Greater than or equal to 1 mg/kg Greater than or equal to 1 mg/kg Greater than or equal to 1 mg/kg
DNA pollen Cq value of less than 36* Table 1. Summary of tests and required results. * The results of an MPI manuka pollen DNA test are reported as a ‘Cq’ value, and for a sample to meet MPI’s standard for manuka honey, the Cq value needs to be less than 36.
If you have test results close to an important threshold, we suggest that you talk with your laboratory about their UoM for that testing method. In general, most methods would have a UoM of at least plus or minus 10%, and it can easily be higher.
• You submit a sample, and get an actual result of 420 mg/kg, which is above the threshold. • However, it falls within the UoM range of 360–440 mg/kg for the method compared with the threshold level of 400, and there is a reasonable chance that your sample could give a result below 400 mg/kg if re-tested.
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MPI MANUKA POLLEN DNA TEST What is the background to the test? MPI’s Mānuka Pollen DNA test works on the principle that bees collecting nectar from mānuka plants will also collect some mānuka pollen at the same time, which will end up in the honey. As pollen grains contain DNA, we can use DNA testing techniques to test honey for mānuka pollen. Real Time PCR – the purified DNA has special reagents added to it, including small sequences of DNA (primers) that have been specifically designed to target DNA sequences that are unique to mānuka, and some DNA probes that have chemicals bound on to them that fluoresce. The sample is loaded onto a PCR instrument, which is pre- programmed to correctly run the test.
What results come from it, and how do I understand them? The results of an MPI mānuka pollen DNA test are reported as a ‘Cq’ value, and for a sample to meet MPI’s standard for mānuka honey, the Cq value needs to be less than 36. To understand this, a brief explanation is needed. Real Time PCR relies on things called ‘primers’ and ‘probes’ that are specific to mānuka DNA, and are added to the sample as part of the PCR reagents. When testing a sample, the PCR instrument goes through a series of cycles, and in each cycle: • The primers and probes attach to any mānuka DNA which is there; and • A copy of the targeted areas of DNA is made; so that • At the end of the cycle there are two times as many copies of the mānuka DNA than there were at the start of the cycle. For example, if there were 10 pieces of mānuka DNA in a sample at the start, there will be 20 at the end of the first cycle; 40 at the end of the second cycle, 80 at the end of the third cycle; and so on. MPI has set a threshold level of mānuka DNA that must be in a sample being tested by PCR, and the Cq value is the number of PCR cycles needed to reach that threshold. The more mānuka DNA in the sample at the start, the fewer cycles will be needed to reach the threshold. If a sample takes 36 or more cycles to reach the threshold (a Cq value of 36), then the original honey sample does not have enough DNA in it to meet MPI’s proposed mānuka definition. A Cq result of 36 is very close to the method’s reporting limit. Because of Uncertainty of Measurement at this level, we suggest that a Cq result of 34.5 or higher is treated as being at risk of failing if re-tested in future.
DNA stands for ‘deoxyribonucleic acid’, and is found in cells of every living organism (including plants like mānuka). The DNA of all plants is actually quite similar; however, different species or varieties of plants have their own unique sequences of DNA, and DNA tests are designed to look for these differences. How is it done? The pollen DNA test uses a technique called Real Time PCR, which is one of the common ways of doing DNA testing. Generally, the test requires about three days to complete from start to finish. The equipment needed is not too expensive (perhaps $100,000 in total). However, the testing process is quite expensive, with a number of manual steps and expensive consumables. Overall, labs are tending to charge $90–$100 + GST per sample. There are five main steps in the testing process: Isolate the pollen from the honey sample – some honey is weighed into a testing tube, and after being dissolved with water the sample is spun in a centrifuge. The pollen forms a pellet in the bottom of the tube as it spins, and the rest of the dissolved honey can then be poured away. Break up the pollen and release the DNA from inside it – after washing the pollen to remove any honey residues, an extraction solution is added. Among other things, it contains very small and hard micro-beads. The sample is put onto a very aggressive shaker (called a bead beater) for a few minutes, and the micro-beads bash the pollen grains and break them open. This releases their DNA into the liquid in the tube. Isolate the DNA – the liquid from the tube is poured through a special ‘affinity’ column which binds any DNA in the sample onto it, while letting the liquid and other things in the sample pass through. After a couple of washing steps, the purified DNA is washed off the column and is available for analysis.
Results are reviewed and approved for reporting – the PCR instrument will produce a result for each of the samples that have been tested. A suitably qualified member of the lab team will review these results, including quality control samples, before approving results for reporting. Quality control Laboratories carry out a number of tests and checks each time that they analyse samples for quality control purposes. In the case of DNA pollen tests, these will include: • Blank QC samples, which should have nothing in them, and therefore check for contamination in the lab. • Control samples, which are honey samples the lab uses repeatedly to check that the method is working correctly. The lab will know what results to expect for these honeys, which are usually chosen to be at the bottom and top of the normal range seen in commercial samples. • Duplicate samples, which may be the same sample tested twice in the same batch, or may be a sample from a previous batch that is re-tested. The purpose is again to confirm that the testing is producing consistent results using a ‘live’ sample submitted by a customer. • An ‘internal control’. Each sample is not only tested for mānuka DNA, but also for DNA found in all plants, to confirm that the testing process has worked for that sample. These general plant DNA tests should work well in all samples—if there is a poor result for the plant DNA markers, it will cause the lab to consider repeating the sample.
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MPI MANUKA CHEMICAL MARKERS TEST What is the background to the test? MPI’s Mānuka Chemical Marker test looks for naturally occurring chemical compounds in honey samples that are also known to be found in mānuka nectar. As honey is primarily made from nectar, these chemical markers confirm that the honey includes enough nectar from mānuka to be called a monofloral or multifloral mānuka honey. Quality control Laboratories carry out checks each time that they analyse samples for quality control purposes. In the case of chemical marker testing, these will include: • Blank QC samples, which should have nothing in them, and therefore check for contamination in the lab. What results come from it, and how do I understand them? Results are reported in milligrams per kilogram (mg/kg) in the honey. One mg/ kg is sometimes called a ‘part per million’ (there are one thousand milligrams in a gram, and one million milligrams in a kilogram). These are the same units used to report other tests that the honey industry may be familiar with—tutin, DHA, MG, HMF, and leptosperin. • Control samples, which include honey samples the lab uses repeatedly, and
How is it done? The Chemical Marker test uses a technique called ‘liquid chromatography–tandem mass spectrometry’ (LC-MS/MS), which is used for other tests such as tutin. The test requires one to two days to complete from start to finish. The equipment needed is quite expensive (over $600,000), but the preparation process is relatively inexpensive in terms of both labour and consumables. Labs are tending to charge $90–$100 + GST per sample. Weigh and dilute the honey sample – after weighing some honey into a testing tube, it is dissolved and diluted with a pre-prepared extraction reagent. This prepares the sample for analysis, putting it in the right condition to be run on the mass spectrometer. Analyse the sample on the mass spectrometer – the diluted honey is put into small vials and analysed on the mass spectrometer. Liquid chromatography is used to separate the various compounds found in the sample, before the concentration of those compounds is measured by the tandem mass spectrometer. All four of MPI’s chemical markers are able to be analysed in the same run through the instrument if the lab wishes to do so. Results are reviewed and approved for reporting – the mass spectrometer produces a result for each of the samples that have been tested, which is reviewed by a member of the lab team. After confirming they are satisfied with the results, and with quality control samples, they are approved for reporting. A summary of the testing process is:
samples which have had known amounts of the chemical markers added to them by the lab. The lab will know what results to expect for these honeys and will check this when reviewing results. • Duplicate samples, which may be the same sample tested twice in the same batch, or may be a sample from a previous batch that is re-tested. The purpose is again to confirm that the testing is producing consistent results using a ‘live’ sample submitted by a customer. • An internal control (or system monitoring compound), which confirms that the preparation and analysis process has gone well. A known amount of this internal control is added to each sample during preparation, and the lab expects to see this in the results for that sample after testing. If the amount of systemmonitoring compound is low (or high), it is an indication that the sample may need to be retested.
To interpret the results for a sample, simply compare them to the levels in MPI’s proposed definition. However, if you have a result that is close to the minimum level required by MPI, there is a chance that a re-test of the honey may see that result change to fall below that minimum level due to the variability in honey samples and in testing methods. See comments earlier in this article. This is particularly important for the three chemical markers that have a required level of 1 mg/kg (which is very close to the method’s reporting limit). In this case, we suggest that a result of 1.5 mg/kg or lower is treated as being at risk of failing if re-tested in future. Left to right: Analytica technicians Luca and Kris with the AB Sciex 6500 mass spectrometer, used for analysis of compounds like tutin and MPI chemical markers in honey. Photo courtesy of Analytica Laboratories.
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VISIT TO BAYER BEE CARE: MONHEIM AM RHEIN, GERMANY OUT AND ABOUT Don MacLeod Over the past few years I have had the opportunity to meet with Dr Christian Maus, Lead Scientist of Bayer Bee Care, when he has been in New Zealand. Dr Maus has repeatedly invited me to visit the Bayer Bee Care Center in Germany.
A private trip to Europe for three weeks in April meant that I could visit the Bayer Bee Care Center on 2–3 May before returning home. This is a report of that visit. The Bayer Bee Care Center is located on a 160-hectare campus at Monheim am Rhein in Germany, just a few kilometres south of Dusseldorf. This site includes the worldwide headquarters of Bayer Crop Science and Bayer Animal Health. For Bayer Crop Science, it is the centre of their insecticide and fungicide research and development (herbicides are located at another site). Meetings with staff My first meeting was with Dr Christian Maus and Coralie van Breukelen-Groeneveld, Head of the Bee Care Center. Our discussion covered their activities and purpose and what is happening in the wider world with respect to pesticides and their effects on bees. The Bee Care Center at Monheim was opened in 2012 to help build a transparent and collaborative platform to promote bee health. Bayer Crop Science and Bayer Animal Health have recognised that they needed a complete understanding of their products and the health of pollinators, especially honey bees. Bayer believes that the challenges faced by beekeepers keeping bees healthy are multifactorial; i.e., there is no one factor that causes all honey bee problems. The factors include varroa, control of pathogens, a healthy food supply and environment for bees. It costs Bayer up to US $100 million (plus 11 years’ research and development) to bring a new pesticide to market, and companies now need to assure pollinator safety. Bayer has identified that there is increasing public interest in not just honey bees, but
Tunnel house research trial at Bayer’s Hofchen Research Station, 2 May 2017. Bayer is trialling a new spraying technique on flowering canola. Note the nuc in the right-hand corner of the tunnel house. The methodology is described in the OECD field test 75 Honey Bee (Apis mellifera L.) Brood Test under semi-field conditions. From left to right: Volkmar Krieg, Peter Trodtfeld, Coralie van Breukelen-Groeneveld, Dr Ulrich Krieg.
many countries worldwide. Bayer wishes to be transparent in its research efforts and encourage open discussion. The company has even had groups opposed to pesticides tour their facility at Monheim and talk to the researchers. They do not anticipate that the European Union will lift the ban on certain seed treatment uses of clothianidin and imidacloprid soon. Their personal opinion is that it is now a political matter and expect that the ban will remain in place. I asked them what they would see with the Bayer takeover of Monsanto, in particular Monsanto’s patent on the use of RNA interference (RNAi) techniques to control varroa. They did not expect the takeover would occur until 2018 at the earliest and there had been no discussion of this patent at their level. I was concerned it had blocked
bumble bees and solitary bees. Both the public and media are asking questions about their safety and welfare. Bayer has recently extended its Bee Care research programme and has now commenced research testing of pesticides against honey bees, bumble bees and solitary bees. Note that no country has yet mandated pesticide testing on bumble bees and solitary bees as a registration requirement to date. I find it very refreshing that a corporation is conducting pure environmental research for a future unknown use, whereas our Government is focusing research only on probable outcomes with forecast positive benefits. A major purpose of the Bee Care Center is to share their research on honey bees with as large an audience as possible, so Bayer has rolled out its Bee Care programme in
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research around the world that could have helped beekeepers. They said they were aware of at least two German universities already researching the use of RNAi to control varroa. Bee testing and risk assessment in ecotoxicology I next met with Dr Maria Teresa Almanza. Her group is part of the Environmental Safety work that is conducted on site. Dr Almanza has nine staff and a dedicated lab conducting bee testing for Tier 1 studies. Her team is also doing work on testing bumble bees (bombus terrestris) and solitary bees using the red mason bee (Megachilidae osmia spp). She describes bees as “little biological black boxes” that are just beginning to teach us how they function. Currently the team is also doing a lot of work renewing pesticide registrations for the European Food Safety Agency, which means ecotoxicity testing of honey bees. Dr Almanza also said that Bayer was now testing the tank mixture as per the label recommendation, which I welcomed as it includes spray tank adjuvants. We toured Dr Almanza’s laboratory where they showed me their present methods for complying with OECD Guidelines. We then discussed the limitations they had experienced. They showed me their testing methods for the OECD 213 Acute oral and OECD 214 Dermal Test and the brood test for honey bees.
• bumble bees: larvae testing is not possible as the bumble bee caps the larvae cell, opens it for feeding and then recaps it. Note that bumble bees always lay their eggs only in clean new wax cells. Varroa control Bayer has its varroa control products business in the Bayer Animal Health division. I met Dr Klemens Krieger, Head of Special Projects, Bee Health. Bayer is looking for new acaricides suitable to control varroa, but none have been discovered to date. A key issue is that all treatments have some adverse side effects, and at low rates to protect bees they do lose some effectiveness. The other issue is that the worldwide market for varroa control products is very small when compared with other animal health markets, so it is a challenge to get resources. Dr Krieger confirmed that later in 2017, Bayer is going to release the Bayer PolyVar Yellow 275 mg beehive gate i . This is an acaricide embedded in a polymer entrance gate, which the bees pass through on entering the hive and are coated with flumethrin to control adult varroa. Results to date have shown excellent varroa control for periods up to five months. Dr Krieger does not know at what stage market release will occur in New Zealand, but he expects it will happen. Bayer had hoped to release three beehive gate products, one containing flumethrin and one of which was to contain coumaphos in order for beekeepers to have three products to manage resistance to any one chemical. Unfortunately, they have some challenges continued...
We discussed the limitations of the dermal test, as it does not represent actual pesticide spraying and is of academic interest only. The OECD field test 75 Honey Bee (Apis mellifera L.) Brood Test under semi-field conditions in a tunnel house (see photo on previous page) has proven to be very reliable based on Bayer’s global experience. Bayer had been experiencing problems with the OECD guidelines for chronic pesticide testing of brood. They were experiencing high larvae death rates at day 8 with no explanation, and the test was not providing consistent results when subject to multiple testing using the same materials. Bayer expressed concerns about the method and test design, and feels that the fragility of the egg and larvae are contributing to their problems. They will continue the testing but expect major changes for the method. Their goal is to develop a robust modelling approach to simulate hive, crop and environment. They are struggling with Tier 3 studies as it is impossible to reduce severe disruption of the hive. As for the other pollinators, the bee tests are not often suitable. Examples include: • red mason bee: acute oral testing is not possible, as the bee can choose not to feed for some days. Larvae testing is not possible, as the bee seals the egg with pollen and nectar sufficient for larvae growth and development and never
reopens the cell until the bee hatches. Also, the red mason bee lays cells on top of each other: the first laid is the last to hatch
Bayer’s Chief Beekeeper Hivemaster Peter Trodtfeld with the old apiary house at the Hofchen Research Farm. The apiary house is no longer used.
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NEW ZEALAND BEEKEEPER, JUNE 2017
developing a polymer to hold and release coumaphos. I advised them that New Zealand beekeepers were adverse to using coumaphos due to honey residue concerns. Bayer’s hivemaster I had a very good afternoon with Peter Trodtfeld, the company’s chief beekeeper. He has a number of hives on the Monheim site, some managed by himself and others by commercial beekeepers, along with his own office and hive workshop and honey extraction setup. Peter took me to Hofchen, the oldest Bayer Research Farm ii , which they use for semi-field studies to investigate pollinator safety of their products. Dr Ulrich Krieg, head of the Experimental Research Station, showed us around the site. As alluded to earlier, we inspected tunnel houses set up over flowering canola to test a new spraying technique that was designed to prevent bee losses during flowering. These tunnel houses each contained a nuc-sized hive. One was an unsprayed control and the other tunnel houses had single but different treatments. The research station also had a traditional apiary house that was used in Europe for housing bees (see photo on page 17).
Left to right: Dr Ralf Nauen (Principal Scientist, Research Insecticides), Don MacLeod, Dr Christian Maus (Global Lead Scientist Bee Care). Photos supplied by Don MacLeod.
Bee toxiogenomics Have you considered how our honey bees can ingest toxic nectar and survive? The best example is our naturally occurring toxins, tutin honey dew from the tutu bush, which a honey bee ingests and then forms honey in the hive. Tutin honey dew has no effect on the honey bee, but karakin nectar from the karaka tree is highly toxic to honey bees— they do not make it back to the hive. What is going on within the bee?
Dr Ralf Nauen, Principal Scientist, Research Insecticides has been studying how Bayer can protect bees with comparative toxicology and functional genomics through The Toxicogenomics Project. Dr Nauen started with the observation that honey bees are not affected at all by the naturally occurring insecticide nicotine and the neonicotinoid insecticide thiacloprid, which has a very low toxicity to bees iii . But the neonicotinoid imidacloprid is 2300 times
Peter Trodtfeld’s Bayer Bee Care Center (flat roof building) with hives.
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Communicating to beekeepers I met with Gillian Mansfield, the
more toxic to honey bees than thiacloprid when LD50 acute values are compared. Not all neonicotinoids are equal in their effects on bees. Dr Nauen’s team of researchers examined the 46 P450 cytochrome enzymes in the gut of the honey bee and have identified a single enzyme that rapidly neutralises the thiacloprid molecule by adding an OH molecule to the chemical. Within two hours of exposure by the honey bee to the pesticide, it has been almost completely neutralised. This enzyme is not present in bumble bees or solitary bees, which means they are susceptible to thiacloprid poisoning. The enzyme is not active against the imidacloprid molecule, so it remains highly toxic to all species. The Bayer team has identified the CYP genes that form the cytochrome P450 enzymes in the three bee types. Dr Nauer and his team have gleaned a great deal of information on how a bee can ingest a toxic substance and the reasons they can manage that substance safely or die. So what is this research leading to? Dr Nauen believes that Bayer will be able to eventually model the effects of different pesticides on beneficial insects, develop pesticides that show complete safety to honey bees and further our understanding of how insects grow resistance to insecticides. One day, we hope that someone in New Zealand will commence a similar study on tutin and karakin toxins.
Bayer now has 20 sustainable agriculture projects worldwide that involve pollinators. In Eastern Europe, Bayer is encouraging the planting of wild flowers (annuals) to diversify pollen and nectar sources for honey bees and bumble bees. I discussed our Trees for Bees programme. Juliana admitted planting trees was a very good idea, so I have agreed to put them both in contact with Dr Linda Newstrom-Lloyd. Overall I was very impressed with the scientists working on the Bee Care team at Bayer. It is fair to say that Bayer are addressing a public relations issue with their Bee Care Centers, but they are backing this effort up with good science. They are working extremely hard to understand more about our number one pollinator, the honey bee, and are also researching other pollinators to understand their role in agriculture. This is a major investment in all our pollinator’s future for which Bayer’s efforts should be acknowledged. Disclosure My travel expenses to and from Europe and accommodation in Monheim were at my expense. I was in Europe for other activities and tacked this visit onto the end of my trip. I received some local transport assistance from Bayer, such as hotel pickups and local transport only and they shouted me local meals. Bayer has not imposed any confidentiality requirements, nor has it checked this report in advance of publication.
Communication Manager at Bayer Bee Care Center. She is looking at how Bayer can best communicate with beekeepers, especially with their global programmes ‘Feed a Bee’ and ‘Healthy Hives 2020’, which is being run in the USA and now launched in Chile. We discussed our colony loss survey in New Zealand as well as the COLOSS data in Europe and elsewhere. Sustainable agriculture Dr Juliana Jaramillo, Scientist Bee Care is working on pollinators and sustainable agriculture. In a study of melon growing in Brazil, Bayer observed that 450,000 hives were used to pollinate 12,000 hectares of melons. The only reason that so many hives were being used was that the hives were very weak and had poor populations of worker bees. The challenge for sustainable agriculture in these areas is to have trained beekeepers who know how to manage healthy hives so less are required. Brazil also has native pollinators, solitary and social, and no one knows how useful these could be in pollinating various crops. Research on non- Apis species has just started and Bayer wishes to understand how they interact with Apis mellifera. Do honey bees displace native solitary bees when foraging an area? Bayer is working with the help of guidelines from the IPBES iv reporting network.
i https://beecare.bayer.com/media-center/news/detail/a-new-way-of-protecting-bees-against-varroa-mites ii https://beecare.bayer.com/media-acenter/beenow/detail/hoefchen-estate-field-trial-station-ensuring-crop-protection-products-do-not-affect-bees iii https://beecare.bayer.com/media-center/news/detail/thiacloprid-is-safe-for-bee-colonies iv http://www.ipbes.net/ The Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services (IPBES)
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