Investigating the Antimicrobial Properties of Scottish Honey: A Citizen Science Approach

Lead Research Organisation: University of Edinburgh
Department Name: The Roslin Institute

Abstract

Honey and other honey bee products have been used as medical treatments throughout history. Following research carried out from the 1970s, and an ongoing and highly successful marketing campaign by the New Zealand Government, manuka honey commands a premium price due to its 'proven' medical benefits. Nearly all of these benefits are due to the strong antimicrobial activity of manuka honey, which are measured in terms of unique manuka factor (UMF).

A valuable industry has grown up around the medical benefits of manuka honey and it is the only honey currently used in human and veterinary medicine for wound healing. Wounds in humans and animals are often painful and healing may be delayed and be prone to complications, which are often due to infection. In medical and veterinary practice honey is routinely used, and dressings that contain medical-grade manuka honey have proven effectiveness for treating complicated wounds in people and animals.

Due to the high cost and limited availability of manuka honey, there is interest from vets and doctors in identifying other sources of honey for use in wound healing, which could be used in both developed and low and middle income countries (LMICs). The ability to effectively treat wounds in the estimated 100 million working horses, donkeys and mules in LMICs using cheaper or even locally produced honey would create a huge improvement in animal and human welfare and significant socioeconomic benefits.

Researchers at the Royal (Dick) School of Veterinary Studies have investigated the antimicrobial effects of a range of honey from across the world, and a surprise result was that honey produced by bees foraging on Scottish heather plants were as effective at inhibiting the growth of common wound microbes as manuka honey. A number of other studies are beginning to support this observation, and we would like to investigate a wider range of Scottish honey, sourced and tested with support from a team of citizen scientists, to confirm the potential antimicrobial activity of heather honey.

The citizen scientists involved in the project will come from beekeeping associations and secondary schools with apiaries from across Scotland, and will take part in a two-day, laboratory-based workshop at the Easter Bush Science Outreach Centre at the University of Edinburgh. We will use and compare different experimental methods to test the antimicrobial properties of honey samples from their own hives and control samples, and also analyse the moisture and pollen content of the honey samples. The results of these experiments will create a new set of data that can be used by scientists and clinicians at the University of Edinburgh to plan further citizen science and research projects.

The citizen scientists will also provide feedback on a new educational toolkit "Honey for Horses", created as part of the project to support secondary school teachers to develop investigations with their pupils into the antimicrobial properties of honey. This toolkit will be made freely available online at the end of the project for use in secondary schools.

Technical Summary

Honey samples from citizen scientists will be cultured aerobically prior to the workshop to check for microbiological contamination. Any contaminated honey will be excluded and replacement samples obtained where possible.
Citizen scientists will be sent details of three methods for testing the antimicrobial properties of honey (honey agar assay, disc diffusion assay, well diffusion assay, all including commercial honey and sucrose solution controls) before the workshop. On workshop day 1 they will be supported by the project team to consider each method and design their experiments, ideally comparing all three methods. A commercial laboratory strain of E. coli will be used for all assays (DH5alpha).
1) Honey Agar Assay: Nutrient agar plates containing honey at a range of concentrations inoculated with bacterial suspension and incubated at 37C overnight. Presence or absence of bacterial growth recorded.
2) Disc diffusion assay: Sterile 8.2mm diameter absorbent discs soaked in a range of honey concentrations placed onto inoculated Nutrient agar plates and incubated at 37C overnight. Zones of inhibition measured around each disc.
3) Well diffusion assay: 8.2mm wells bored into the surface of inoculated agar plates and 180ul diluted honey at a range of concentrations placed in each well. Plates incubated at 37C overnight and zones of inhibition measured around each well.
Citizen scientist honey samples and controls will also be commercially tested for quality, including diastase and hydroxymethylfurfural(HMF) levels, pH and water content. Antimicrobial activity will be assessed in terms of total activity (TA) and non-peroxide activity (NPA). The pollen in all samples will be analysed to determine floral source and manuka samples will be tested for Leptosporin for authentication.
Citizen scientists will extract pollen from each honey by centrifugation and prepare microscope slides using fuchsin stain. Moisture content will be measured using a refractometer.

Planned Impact

The project will improve the scientific literacy of the citizen science cohort, especially any beekeepers and teachers without a formal scientific background. It will also bring all participants up to date with current research into the role of honey in wound healing, honey bee health and genetic diversity and give them an opportunity to meet and interact with the scientists and clinicians involved. We anticipate that by choosing to focus on bees, an area of considerable public interest, and revealing the scientific processes required to investigate the role of honey in wound healing via the citizen science workshop and educational toolkit, we will improve public trust in and understanding of both science and scientists.

Via pairing secondary schools with local beekeeping associations and the educational toolkit, we expect to increase the sustainability of school apiaries and encourage young people to aspire to careers in science and beekeeping, in particular highlighting apprenticeship opportunities in both areas and encouraging young people from a range of backgrounds to consider these options.

The project funding will enable the addition of a citizen science strand to the existing portfolio of public engagement with research (PER) at The Roslin Institute and EBSOC, realising a long-held ambition to include participatory research in our PER activities. The experience gained by the project team research and public engagement staff will provide a blueprint for future PER projects and insight into how a citizen science approach may be incorporated into other areas of Roslin Institute research.

The only source of medical grade honey currently available is manuka honey from New Zealand. This expensive, finite and ultimately unsustainable resource cannot continue to supply the medical device (dressings) market indefinitely. As our population ages, and the prevalence of individuals with traumatic wounds and wounds associated with diseases such as type 2 diabetes increases, the demand for honey-impregnated devices, and consequently other sources of medically active honey, is only likely to increase. Assuming this project demonstrates and strengthens the findings of previous work, Scottish heather honey could be an alternative source of medical grade honey for the future. This would have significant benefits for the Scottish beekeeping industry, tourism, and for human and animal health.

Publications

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Description This project was designed to test the antimicrobial effects of Scottish heather honey with a cohort of school teachers and beekeeper citizen scientists through a series of in person workshops. The project was due to start in 2019 but was severely disrupted by the COVID-19 pandemic. An introductory online project launch workshop took place in November 2020 to introduce the Roslin scientists, bee keepers and school teachers.

Our planned in-person two-day workshop for our cohort of teacher and beekeeper citizen scientists (CS) was not possible in 2021 due to COVID-19 restrictions, but we outlined details of an alternative plan to deliver the project online by 31 July 2021, delivering two online workshops in May and June 2021. Experiments were performed between the two workshops by project team researchers at the Roslin Institute, with interaction and feedback from citizen scientists (CS) at their schools and beekeeping associations.

The first of the two half day online workshops for citizen scientist project participants (beekeepers and teachers from across Scotland) was held online in May 2021. Short talks were given by project team members on the antimicrobial properties of Scottish honey, and pollen analysis in honey, and followed by opportunities for questions and discussion. Participants then split into two working groups to discuss and plan the experimental work that the project team would carry out on their behalf before the next workshop (the experimental work would have been carried out by the citizen scientists themselves in the original, pre-COVID-19 plan for the workshop). Each group focused on a different experimental protocol, to determine the best approach for analysing the antimicrobial properties of their honey samples. A third protocol was saved for a catch-up session for participants not able to join the live online workshop. This process revealed how researchers use the scientific method, and required participants to consider all elements of experimental design, including controls, variables and health and safety. Most of the participants did not have a formal science background, and many reported the experience as a great insight into how science works, as well as feeling a sense of achievement having contributed to the design process. How to prepare pollen slides from honey for microscope analysis, was demonstrated, which generated a lot of interest and questions from the particpants, who were very excited at the prospect of carrying out their own experimental work. After the workshop, participants were sent a microscope and other equipment and consumables to enable them to carry out this aspect of the practical work themselves, sharing their results via our online MS Teams platform. The session provoked much lively discussion and conversation, and participants kept in touch with the project team and each other afterwards via the Honey for Horses MS Teams online platform. This enabled them to share questions and advice, and the results of their at-home pollen analysis, which again promoted further discussion and sharing of existing and new-found expertise.

The second and final half-day online workshop for citizen scientist participants and project team was held in June 2021. The session started with an expert analysis of the participants' own pollen analysis images, shared via the Honey for Horses MS Team, and presentation of the results from the commercial pollen analysis carried out by Minerva Scientific. Both of these prompted many questions, comments and a lively debate about apparent inconsistencies between the two methods. Participants also shared their experiences of using the at home/school pollen analysis kits, which were mostly very positive but were not without some challenging moments. In the next part of the session the results of the honey agar, liquid culture and disc diffusion experiments the participants had designed were described, again with opportunities for questions and discussion. The results indicated that the liquid culture method was the most reproducible, and it clearly showed that a significant number of the participant's honey samples had significant antimicrobial activity. Results of the commercial analysis, which indicated that this activity was due to hydrogen peroxide and was not comparable with the activity resulting from Unique Manuka Factor.

Following the workshops, feedback from the citizen scientists was overwhelmingly positive, with all participants reporting that they strongly agreed or agreed that the project had achieved its aims. These were to provide an enjoyable experience for participants; improve participants' scientific knowledge; bring participants up to date with current science and clinical practice related to honey and wound healing; give participants the opportunity to meet and interact with scientists and clinicians; and involve participants in the process of experimental design.

Holding the online introductory sessions and providing an interactive MSTeams space for materials and conversations related to the project meant that we had additional time and interaction with the CS beekeepers and teachers. These online interactions turned out to be very valuable which we wouldn't have realised had the project not been disrupted by COVID-19. Given their success for this project for future projects of this type we are likely to include online introductory sessions and an interactive online space.

Even with the delays and adjustments to the project there has been real enthusiasm from the CS beekeepers and teachers to engage with the activities that we planned for the project in the analysis of pollen and the microbial content of honey and in the development of the honey for horses toolkit. This was made particularly clear in how engaged the participants were in the online launch we had for the project in November 2020 and emphasises how powerful Citizen Science projects like this can be in engaging interested stakeholders.

We also found that having a contact point within the community helped greatly with encouraging participation in the project. In our case Mark Barnett is a scientist and also a bee keeper which helped to facilitate the link between participants, scientists and the bee keeping community. The project was also helped substantially in linking with schools and the bee keeping community by other ongoing projects involving bees within the Easter Bush Science Outreach Centre (EBSOC).
Exploitation Route The final part of the workshop was an exploration of which elements of the project should be used to create the proposed schools toolkit, including whether the pollen analysis experiment should be included, what sort of extra support might need to be included for teachers, and whether there were any non-science curriculum links that could be included. Both teacher and non-teacher participants made valuable contributions, and an additional outcome was the formation of a teacher advisory group to support the development of this resource.

Through participation in the project the beekeepers have expanded their understanding of the potential of the honey they produce for clinical treatment of wounds in horses. They may take this knowledge forwards in diversifying the market for their product. The results of the commercial analysis of hydrogen peroxide activity prompted much impassioned debate, and a productive discussion about how this claim could be investigated further. Participants identified that the next experiment should involve pre-treating the honey samples with a peroxidase and repeating the liquid culture, something that is outside the scope of this project, but has the potential to be a future undergraduate research project.
Sectors Agriculture, Food and Drink,Education

 
Description The project has given the teachers and beekeepers an insight into the clinical uses of honey to treat wounds in horses and insight into research at the Roslin Institute and R(D)SVS into honey, bee health, and genomics. The teachers can see how they could use this information for educational purposes. The Citizen Science beekeepers have new knowledge about the scope and potential value of heather honey. The final part of the workshop was an exploration of which elements of the project should be used to create the proposed schools toolkit, including whether the pollen analysis experiment should be included, what sort of extra support might need to be included for teachers, and whether there were any non-science curriculum links that could be included. Both teacher and non-teacher participants made valuable contributions, and an additional outcome was the formation of a teacher advisory group to support the development of this resource. Through participation in the project the beekeepers have expanded their understanding of the potential of the honey they produce for clinical treatment of wounds in horses. They may take this knowledge forwards in diversifying the market for their product. Following the workshops, feedback from the citizen scientists was overwhelmingly positive, with all participants reporting that they strongly agreed or agreed that the project had achieved its aims. These were to provide an enjoyable experience for participants; improve participants' scientific knowledge; bring participants up to date with current science and clinical practice related to honey and wound healing; give participants the opportunity to meet and interact with scientists and clinicians; and involve participants in the process of experimental design.
First Year Of Impact 2020
Sector Agriculture, Food and Drink,Education
Impact Types Cultural,Societal

 
Description Bee-ology 
Organisation Science and Advice for Scottish Agriculture
Country United Kingdom 
Sector Public 
PI Contribution Research staff, the Easter Bush Campus Apiary team and the Public Engagement team at the Roslin Institute were co-founders of Bee-ology in 2018, bringing scientific, beekeeping, and education and engagement expertise to the collaboration, which aims to get Scottish high school pupils hands-on with real-life bee science. Via the facilities at the Institute's Easter Bush Science Outreach Centre we run hands-on workshops for teachers and pupils, providing access to molecular biology techniques and specialist equipment including PCR, gel electrophoresis and microscopy, as well as scientific staff who support all workshops. This has enabled pupils to carry out microscopic and molecular surveys for two species of Nosema parasite, currently a major threat to both commercial and hobby apiaries. We have also supported the partnership to grow from two to three high schools by inviting teachers from the founder schools and a new school in our UKRI-funded project "Investigating the Antimicrobial Properties of Scottish Honey: A Citizen Science Approach", building valuable new relationships within the partnership. In addition we have helped the partnership to secure ongoing project funding from the Royal Society.
Collaborator Contribution Our lead collaborators at the three high schools involved in Bee-ology - all teachers involved in running their school apiary - have played a key role in engaging and involving their pupils in the network, collecting samples, carrying out investigations at school, and bringing their pupils to the Easter Bush Science Outreach Centre. They have supported our researchers and public engagement team to provide scientific information at the appropriate level for their learners, and contributed new ideas and research questions to the projects we have run together. They also played a key role in our UKRI-funded project "Investigating the Antimicrobial Properties of Scottish Honey: A Citizen Science Approach". Also involved in Bee-ology are colleagues from SASA (Science and Advice for Scottish Agriculture), who contribute their expertise in entomology, bee diseases and bee farming to the partnership.
Impact Recruitment of teacher beekeepers to the UKRI-funded Honey for Horses Citizen Science Exploration project Initial and follow-on funding from the Royal Society Bee science schools workshops at the Easter Bush Science Outreach Centre for Beeology partnership. Creation of standalone Bee Science workshop, bookable by any school visiting the Easter Bush Science Outreach Centre
Start Year 2018
 
Description Scottish Beekeepers Association 
Organisation Scottish Bee Keepers Association
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution Mark Barnett started a Citizen Group with the SBA. The 'Investigating the Antimicrobial Properties of Scottish Honey: A Citizen Science Approach project' is helping to enhance the activity of this group. For this project we are working with them to increase the value of heather honey.
Collaborator Contribution The SBA helped with recruitment to the project and provided a platform to publicise the project, through the The Scottish Beekeeper magazine and through their wider networks and activities.
Impact This collaboration resulted in funding of the project 'Investigating the Antimicrobial Properties of Scottish Honey: A Citizen Science Approach'.
Start Year 2017
 
Description British Bee Keepers Association Magazine Article 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Jed Russell a member of the Honey for Horses team published an article entitled 'Scottish Honeys - Could they have medical applications alongside Manuka?' in the British Bee Keepers Association Magazine. The article described how Jed was involved in one of Roslin's most recent and exciting undertakings - the Honey for Horses Citizen Science Project, funded by a grant from UK Research and Innovation (UKRI). The workshops themselves and the engagement with schools and the beekeepers were detailed in the article as well as the overall outcomes and next steps.
Year(s) Of Engagement Activity 2022
 
Description Honey for Horses Online Project Launch November 2020 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Four teachers and seven beekeepers attended the online project launch. Nicola Stock, Emily Clark, Tim Regan, Patrick Pollock and Mark Barnett introduced content related to the project. Mark Barnett did a live demonstration of laboratory testing of honey for microbial contamination. Attendees listened and had the opportunity to contribute and engage with the project. Due to the ongoing Covid19 pandemic plans for the two day live workshop has been adapted to a two half day on-line workshops in May and June 2021.
Year(s) Of Engagement Activity 2020
 
Description Honey for Horses Online workshop 1: April 2021 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Study participants or study members
Results and Impact The first of two half day (four hours) online workshops for citizen scientist project participants (beekeepers and teachers from across Scotland). Short talks from project team member Dr Patrick Pollock about his work on the antimicrobial properties of Scottish honey, and guest speaker Matthew Richardson on pollen analysis in honey, were followed by opportunities for questions and discussion, hosted by Dr Nicola Stock and Dr Emily Clark.
After a short break, participants split into two working groups to discuss and plan the experimental work that the project team would carry out on their behalf before the next workshop (the experimental work would have been carried out by the citizen scientists themselves in our original, pre-COVID-19 plan for the workshop). Each group focused on a different experimental protocol, to determine the best approach for analysing their honey samples. A third protocol was saved for a catch-up session for participants not able to join the live online workshop (see below). This process revealed how researchers use the scientific method, and required participants to consider all elements of experimental design, including controls, variables and health and safety. Most of the participants did not have a formal science background, and many reported the experience as a great insight into how science works, as well as feeling a sense of achievement having contributed to the design process.
Participants then reconvened for a demonstration by Dr Mark Barnett of how to prepare pollen slides from honey for microscope analysis, which generated a lot of interest and questions from the particpants, who were very excited at the prospect of carrying out their own experimental work. After the workshop, participants were sent a microscope and other equipment and consumables to enable them to carry out this aspect of the practical work themselves, sharing their results via our online MS Teams platform.
This was followed by a presentation from Dr Tim Regan of the results of the initial screening that we carried out on the honey samples submitted by the citizen scientists, and the session closed with information about next steps for the project.
The session was recorded for participants not able to attend the live online session, and an additional online session was run to enable these participants to plan a set of experiments using the third experimental protocol.
The session provoked much lively discussion and conversation, and participants kept in touch with the project team and each other afterwards via the Honey for Horses MS Teams online platform. This enabled them to share questions and advice, and the results of their at-home pollen analysis, which again promoted further discussion and sharing of existing and new-found expertise.
Year(s) Of Engagement Activity 2021
 
Description Honey for Horses Online workshop 2: June 2021 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Study participants or study members
Results and Impact Second and final half-day online workshop for citizen scientist participants and project team. The session started with an expert analysis by guest speaker Matthew Richardson of participants' own pollen analysis images, shared via the Honey for Horses MS Team, and a presentation by Dr Emily Clark of the results from the commercial pollen analysis carried out by Minerva Scientific. Both of these prompted many questions, comments and a lively debate about apparent inconsistencies between the two methods. Participants also shared their experiences of using the at home/school pollen analysis kits, which were mostly very positive but were not without some challenging moments.
The next part of the session was led by Drs Mark Barnett and Tim Regan, who took participants through the results of the honey agar, liquid culture and disc diffusion experiments they had designed, again with opportunities for questions and discussion. The results indicated that the liquid culture method was the most reproducible, and it clearly showed that a significant number of the participant's honey samples had significant antimicrobial activity. Dr Patrick Pollock then revealed the results of the commercial analysis, which indicated that this activity was due to hydrogen peroxide and was not comparable with the activity resulting from Unique Manuka Factor. This prompted much impassioned debate, and a productive discussion about how this claim could be investigated further. Participants identified that the next experiment should involve pre-treating the honey samples with a peroxidase and repeating the liquid culture, something that is outside the scope of this project, but has the potential to be a future undergraduate research project.
The final part of the workshop was an exploration led by Dr Nicola Stock of which elements of the project should be used to create the proposed schools toolkit, including whether we should include the pollen analysis experiment, what sort of extra support we might need to include for teachers, and whether there are any non-science curriculum links that could be included. Both teacher and non-teacher participants made valuable contributions, and an additional outcome was the formation of a teacher advisory group to support the development of this resource.
Following the workshop, feedback from the citizen scientists was overwhelmingly positive, with all participants reporting that they strongly agreed or agreed that the project had achieved its aims. These were to provide an enjoyable experience for participants; improve participants' scientific knowledge; bring participants up to date with current science and clinical practice related to honey and wound healing; give participants the opportunity to meet and interact with scientists and clinicians; and involve participants in the process of experimental design.
Despite the challenges of converting the project from an in-person to online format, participants reported that the "online process was well-designed and executed" and told us "congratulations for delivering such an excellent course. The onset of Covid and the many changes you had to make in order to deliver this Citizen Science Project must have been extremely challenging. However, for me, the participant, it ran like clockwork. You turned many negatives into positives in the way the course was successfully and creatively delivered."
Participants reported that they really enjoyed the opportunity to carry out their own pollen analysis experiments on their honey samples, using microscopes and other materials posted out to them by the project team. A beekeeper participant reported that "making the pollen slides and learning more about my honey" was the best part of the project for them, and a teacher participant said that their students "were delighted to see the various shapes and sizes of pollen". All of the equipment purchased to enable this analysis was loaned out and later returned to the project team, building our capacity to run this activity with participants in future bee science engagement activities. It also acted a proof of principle that complex and valuable equipment could be provided to members of the public in this way, and used successfully without direct supervision.
An unexpected but very positive outcome of participants carrying out this analysis at home or at their school was the involvement of a large number of additional people in this element of the project, including relatives and flatmates, pupils, and in one case families from the wider community via a community engagement event run at a participant's beekeepers association.
Feedback from participants indicated that the project had communicated its science content effectively, with comments including "knowledgeable speakers and researchers - they are able to clearly articulate the science behind the honeys antimicrobial mechanisms" and "The team made us very welcome, the whole event was inclusive and informative. The people involved were extremely professional but never left the non-scientists behind. They took great care to make the information given accessible to all and were always very to answer any questions and received suggestions."
Participants were keen to share their new-found knowledge with others, and all expressed an interest in taking part in future science engagement projects on topics including bee health, climate change and bee breeding, as well as the antimicrobial effects of Scottish honey.
They appreciated "the inclusion, the quality of the information and professionalism of everyone on the team and the opportunity to take part." "Thank you so much for giving me the opportunity of joining the project. Everyone has been so welcoming and encouraging. It was really nice to feel part of a research team and making decisions on the design of the experiments being carried out."
Year(s) Of Engagement Activity 2021
 
Description Honey for Horses online experimental design catch-up session: May 2021 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Study participants or study members
Results and Impact An additional online session held after Honey for Horses workshop 1, to enable participants not able to attend that workshop to be involved in the experimental planning process. Drs Mark Barnett, Tim Regan and Nicola Stock facilitated the session, which focused on designing an experimental approach to analyse the antimicrobial activity of honey samples in liquid culture. Only one of the three expected participants, a history teacher who has set up a new apiary at their high school, attended the session, and they were very enthusiastic about the task in hand despite their lack of scientific background. They reported really enjoying the opportunity to get an insight into the scientific process, and felt a large sense of personal achievement having contributed in such a significant way to the design of one-third of the project's experimental work.
Year(s) Of Engagement Activity 2021
 
Description News Article for the Scottish Beekeeper Magazine 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Mark Barnett was invited to write an article in the Scottish Beekeeper Magazine. He described the project and expected outcomes. The magazine is read by more than 2000 members of the Scottish Bee Keepers Association.
Year(s) Of Engagement Activity 2021