Reishi for everyone!

Authors: Nico Coallier, Geoffroy Renaud

Prologue

Sharing a passion for several years about mycology and biology in general, Geoffroy and Nico had the idea of ​​applying mycology in various ways to beekeeping. Flavo now produces medicinal honeys in collaboration with FNG, such as Reishi-boosted honey which we will discuss in this article. Together, they also validate the potential of mushrooms as a dietary supplement for bees.

Inspired by an article by Paul Stamets published in 2018 in the journal Nature (see here ), they developed their own Reishi extract and application method. For 2 years now, Flavo and FNG have been testing the effect of feeding Reishi on the strength , presence of disease and winter survival of colonies .

Nico being the principal scientist at Nectar Technologies, he is fortunate to be able to access the Nectar 1.20 platform which allows him to easily collect data and set up experiments. At Flavo, we have a history (2022-2023) of more than 170 hives located on more than 20 independent sites, allowing us to test several things in robust experimental designs. All the hives are monitored with Nectar technology and very detailed monitoring is carried out by Nico during each inspection (i.e. monitoring of the strength of the hive, monitoring of the brood, monitoring of the queen, monitoring of the varroa, monitoring aggressiveness, monitoring of treatments and feeding as well as location of hives and more!).

As for Geoffroy, he is currently continuing his studies on the properties of mushrooms at FNG but also through his doctorate at the IRBV . He also has access to mushroom cultivation greenhouses as well as all the necessary laboratory equipment. Together, they want to advance mycology applied to beekeeping and demonstrate the extent of its potential.

We would have understood that Geoffroy has a passion for mycology, but I still had fun asking him about his last memory of a bee sting. Here is his response:

“I was impressed by watching a video portraying a Zen beekeeper from Tokyo who worked without any protection. Each working session with bees began with a meditation, calibrating one's energies and preparing for the encounter with the bee colonies.

While working at the Champs des Possibles, I realized that beehives had been placed between the railway line and the pedestrian crossing, in semi-shade. When I saw a small swarm heading in my direction, I thought of my Zen beekeeper, coupled with the prejudice that the colony must be well accustomed to visitors and disturbances (being close to the CP rails). Apparently, I hadn't reached the Zen maturity I imagined and I was treated to a good dose of injections on my upper body. Fortunately, I was at least zen enough not to panic and to kindly get rid of the bees as nuisances. Since then, I have filled my class with honey plants and I have been around populations of wild bees and bumblebees without having repeated the unpleasant experience of the sting. It seems that I am continuing my ascent towards the necessary zenitude. “

Good reading!

Who is Reishi?

This mushroom from the polypore family has several names including Reishi but also shiny ganoderma, shiny polypore, mannentake or linghzi . The scientific name of its genus is Ganoderma and includes more than eighty species. The most studied of these species, Ganoderma lucidum, specifically refers to this ligninolytic basidiomycete fungal organism naturally present in Europe and Asia. Archaeological traces demonstrate that this mushroom has been used in traditional medicine for 8,840 years (±44 years) (Yuan et al. 2018).

Photo of Reishi growing in FNG greenhouses.

4 of its closest cousins ​​are found in North America, including Ganoderma tsugae or Hemlock Ganoderma, endemic to Northeast America and therefore hardy in Quebec. While bees most likely forage on the exudates of this Quebec Reishi, we work with the Ganoderma most studied and best documented by contemporary science, that is to say Ganoderma lucidum.

The virtues of Reishi honey

Beta glucans, chitin and ergosterol are central to the cellular development of fungal organisms. Although the presence rates can vary from one species and one genus to another, we recognize here an analytical triptych common to the vast majority of fungal organisms.

What then establishes the particularity of the various fungi used by pharmacopoeias around the world are the molecules specific to the various fungi, spores and molecules of the genera and species under study.

Below, 7 bioactive compounds specific to Ganoderma lucidum have been modeled in studies seeking to determine their interactions with various pathogens as well as its signals and impacts in the host.

Source - Mushrooms and Active Biomolecules

Medicinal mushrooms are full of benefits, associated with various bioactive principles corollary to various stages of organism development. While the scientific literature demonstrates that bioactive principles are unique to each species, a common core allows us to begin to understand the various mechanisms at work.

At the heart of every fungal organism cell is an ergosterol-rich membrane, a chitin cell wall coated with triple-ellipse 1-3, 1-6 beta-glucan. These three bioactive components stimulate the microbiota in their own way and can provide significant quantities of fiber even when solubilized.

Beta glucans also act more specifically as immunomodulators (ie adaptogens), prompting a recalibration of the immune system's response to environmental stress and stimuli, while reducing cholesterol and blood sugar levels.

The particularity of fungal beta-glucans, namely their triple-elliptical shape, significantly accentuates the effectiveness already recognized in plant beta-glucans, as studied in oats. Thus, while beta-glucans 1-3, 1-6 make it possible, for example, to reduce the level of cholesterol in the blood, the maximization of the contact points made possible by the triple-elliptical shape of the fungal compounds makes this action more rapid and more efficient.

Ergosterol is a molecule belonging to the sterol family, and therefore acts as a stimulant in the host. For fungal species, it plays the role of cholesterol in mammalian cell membranes. A particularity of interest of this molecule is that it reacts to light by becoming ergocalciferol, a precursor molecule of vitamin D2, whose health benefits far exceed the assimilation of calcium. The importance of this vitamin in preventing cytosine shocks and therefore reducing the incidence and violence of autoimmune reactions has been extensively studied during the most recent pandemic.

To return to the particularities of reishi, its millennial reputation has long given it the nickname “mushroom of immortality”. Not only does it have an impressive list of significant therapeutic effects, it is also known as an “enhancer” or “tonic” of the beneficial effect of other virtuous herbs and mushrooms. Thus, some schools of traditional Chinese medicine integrate reishi transversally, that is, reishi is integrated into all of their herb and mushroom formulations.

In vivo and in vitro studies as well as population studies support the benefits of reishi on cardiovascular health, the regulation of blood pressure, cholesterol levels and blood sugar levels. Reishi helps delay viral growth, reduces inflammation and supports protection against free radicals and tissue damage. Reishi has also been shown to invigorate immune functions while inhibiting the growth of tumors. It protects and tones the liver while relieving symptoms of anxiety, insomnia and depression. Studies have also shown that regular intake of reishi has a significant impact on improving cognitive functions, energy and respiratory functions. Of course, this list is not medical advice and is in no way intended to replace the advice of a health professional.

For a summary of all the benefits of Reishi, visit the Health Canada monograph or the FNG website .

How is it integrated into honey?

At Flavo, our honey will be churned for 86 hours at very slow speed in order to preserve all its properties and flavor. We always add a little propolis and wax to give the honey even more flavor.

When we prepare Reishi honey we mix the extract at the end of churning and leave to infuse for a minimum of 48 hours before taking away. Unpasteurized honey combined with the umami that Reishi brings is a very surprising delight!!!

It's our best seller and I personally still eat it by the spoonful every day for its benefits but also for its taste!

Only for humans?

As discussed in the introduction to this post, we are also testing the benefits of Reishi in bees. Indeed, we are currently working on a scientific publication but here is a note on our progress in researching the benefits of Reishi in bees.

Bee consuming a Reishi-based solution.

Note that while these natural health ingredients still have a lot to teach us, the scientific literature is already very abundant regarding their remarkable qualities and applications for therapeutic interventions for humans but also for many living organisms.

For example, a recent study demonstrates that when a chitin supplement is offered to bee colonies battling Varroa destructor infestations, the time to remediate a gaping wound can be reduced by 150% to 900% depending on the dosage. (Özkırım et al. 2022).

On the other hand, Stamets and his team demonstrated in 2018 strong effects in reducing the viral dose of virus associated with varroa destructor (e.g. an observed reduction of 79 times at the level of the deformed wing virus) (Stamets et al. 2018).

Additionally, flower nectar is the habitat of complex fungal communities (von Arx et al. 2019; Olaitan et al. 2007). The complexity of the composition of nectar therefore does not come down to sugar water. However, beekeepers feed their hives in preparation for winter with a syrup composed in the majority of cases only of water and refined sugar (i.e. white sugar). The impact of this food intake to avoid mortality due to a lack of reserves during the winter can be significant.

Several positive or negative effects can be induced through diet (Frizzera et al. 2020; Tawfik et al. 2020; Oskay et al 2021; Barker et al 1978; Agarwal et al. 2023). Several additives are present on the market in an attempt to increase the positive impacts of a sugar-based diet (eg: SuperDFM , HiveAlive , etc.). None of its supplements use mushrooms in their recipes despite having certain potential.

Other beekeepers prefer to feed with honey and this is still possible. On the other hand, we must also be careful since autumn honey has higher humidity than honey harvested earlier in the season and some suggest that this can lead to dysentery and a proliferation of bacteria in reserves (Oskay et al 2021 ). We must not forget that the domestic bee is not native to our climate and the shorter harvest periods result in a shorter period for the maturation of the honey and therefore this end of season honey has not matured long enough which could explain this effect. Thus, syrup feeding is generally recommended but is not the only possible method and this is why we will test two methods of applying Reishi.

A first method where we simply added our extract to the end of season feeding then a second method by dripping explained in more detail in the following sections.

We decided to tackle the unexplored potential of Reishi first but we have several studies underway into other mushroom species as well as combinations. In the next sections, we will focus on the effects of Reishi on survival from last winter, the reported effects of treated hives that survived the winter and the immediate effects of the treatment carried out during the 2023 season.

Effect on winter survival

In 2022, we decided to add a Reishi solution to our end-of-season feeding syrup. In fact, beekeepers will feed the hives with a syrup (generally 3:1 at the end of the season) in order to compensate for the short summer season in our region and promote better colony survival.

It was therefore logistically easy to add varying doses of Reishi to the feeding syrups and to analyze the impact on the survival of the hives. We therefore carried out a minimal treatment of the hives against varroa (uniform use of formic acid and a spray of oxalic acid at the beginning of November, two organic products) in order to better quantify the effect of Reishi. Note that we had an unidentified event that resulted in a heavy winter loss last winter. The sample is made up of 113 hives which were wintered in 3 outdoor sites located in Estrie, Quebec.

We then analyzed the data using the CoxPH model. This survival model allows the analysis to avoid bias due to censored data. Indeed, we do not know the date of death of the hives that survived until the end of the experiment, i.e. June 15, 2023. Since some hives could die following this date, we must consider this element in the modeling to avoid to overestimate survival. Below are the survival curves estimated by the model for varying doses of Reishi. Doses consist of the number of feedings in which a supplement was added. The effect of reishi is significant (p-value: 0.01) and increases the probability of survival by a factor of approximately 27% per dose administered. We observe an average survival 13.4% higher compared to the average survival for hives treated with a minimum of one dose of Reishi and a maximum of 4.

Effects carried over to the following season

We wanted to validate whether an effect (or a bias in our analyses) could have been caused by a reported effect. A reported effect would be an effect of a treatment in 2022 which is transposed to the health of the hive in 2023. We therefore identified the hives which survived and which received a Reishi treatment in 2022 then compared with the survivors who did not. not having received treatment.

The dataset includes 31 treated hives that survived versus 4 untreated hives, so the analyzes are only exploratory due to the small sample size. However, no carryover effects were detected. We still observed a slight trend in the rate of varroa fall where the milked hives seemed to show slightly higher rates, suggesting greater hygienic behavior.

Analyzes were performed on reports between the start of the season and August 1, when Reishi treatments in 2023 began. We will continue to evaluate the effects over several seasons in the future.

Immediate effects

In 2023, we took our thinking further. Honeybees show two phenotypes (two different gene expressions), summer bees which live about 40 days and winter bees which can live more than 200 days and which have a much larger fat body (Alburaki et al. 2022).

These winter bees begin to be born at the beginning of August in Quebec. Feeding larvae is essential to good bee health, especially since epigenetics plays a much more important role in bees than in humans. Thus, our hypothesis is that by feeding bees from the beginning of August, we should promote more resilient winter bees and therefore better winter survival.

As the honey boxes are still present at the beginning of August, we had to develop a new application method. Based on the oxalic drip treatment, we made a 1:1 solution into which we infused the Reishi extract. The application of this solution was done at each visit (ie approximately every two weeks) on 50% of our livestock. This allows us to avoid contaminating the honey since we only apply 50ml of Reishi syrup directly to the bees. To mitigate the potential bias of bees cleaning the syrup, we also fed the control hives with a 1:1 solution without Reishi. The hives were chosen randomly in each apiary to reach 50% of each apiary but to avoid effects linked to the strength of the hive or the location. A total of 108 hives made up the sample, including 54 treated and 54 controls.

Application of Reishi by dripping.

Subsequently, we analyzed the data using linear mixed models to detect potential instantaneous effects. We call instant effects, effects observed in the days following an application. We decided to launch this analysis following Nico's observations who noted that the treated hives were visually much stronger and more active. We simply wanted to validate his intuition and we were surprised by the results.

Effect on varroa fall rate

We evaluated the effect of Reishi dripping treatment on the observed varroa shedding rate. We supplemented the control hives with a 1:1 solution without Reishi. For a period of 10 weeks starting August 1, 2023, we supplemented during each visit at ideal intervals of 10 to 14 days but some periods were shorter or longer. We therefore normalized the fall rate over 48 hours (ie count/48h). Varroa mites on the plateau located below the mesh floor of the waterfall were counted. All decks were oiled and cleaned at each visit. Towards the end of summer and autumn, the activity of the hives being less, it remains possible to make an exhaustive and precise count of the varroa found on the plateau.

We expected no effect on varroa fall rate since our initial goal was to treat bees with the aim of affecting their winter survival. However, during the analysis we observed large differences between the groups. Figure 1 shows the change in varroa for hives that were treated (Reishi) or not (Control). The first count following the application of Reishi is generally higher for treated hives of 7.72 varroa on the fall rate over 48 hours (p-value < 0.005). Additionally, Reishi is strongly correlated with a decrease in the rate of falling of -0.2 per week (p-value <0.012). We analyzed the data using a mixed model following a Poisson distribution. We compared several models and selected the most parsimonious.

Progression of varroa depending on the week of the year for the 10 weeks of the experiment.

Effect on hive condition

During each visit, we also counted the number of new frames covered with bees, the number of frames covered with brood, and the number of frames with nectar and pollen reserves. In addition, we identified whether the queen was healthy and noted any abnormalities. No immediate effects were detected in terms of hive strength, brood quality or the presence of the queen. An observation was made by Nico who nevertheless noted more active hives but we did not collect data on this variable.

Effect on the fall harvest

We also counted the number of frames of honey harvested. A full frame is determined to be capped on a minimum of 80% of the frame. Otherwise, we combine incomplete frames when counting and the value is rounded down. Here is the distribution of the fall harvest.

We can see a higher harvest at the level of the hives treated during the month of August, which is consistent with the observations of higher activity despite similar bee populations. Reishi could promote stronger immunity and therefore more active bees! More data and research needs to be done on the immediate effects of treatment since this was not our research objective. Our main objective remains to increase winter survival by reducing the necessary treatments.

Conclusion and next steps

During the winter, FNG will deploy a major update to its analytical laboratory relating to active ingredient extracts. It will thus be possible to refine with more precision the variabilities in the levels of active ingredients present in the extractions of mushrooms, mycelium and spores.

The current studies could be replicated according to various concentrations of multiple active ingredients and thus identify with greater precision the bioactive compounds determining the effects described above. In short, moving from an observational “quantification by input” analysis to quantification by active ingredient. It will then be possible to adjust the formulations according to the various treatments implemented.

We are also calling on beekeepers interested in participating in the study to contact us during the winter of 2023-2024 in order to be a user of the supplement next season. We would like to reach a sample of more than 2500 hives in order to improve the analyzes and better understand how fungi interact with bees.

We will also add a component of analysis of the microbiota of treated bees as well as more experiments in the future in order to continue to validate and develop mycology applied to beekeeping.

Contact us if you are interested in participating or if you have any questions :)!

References

HOBBS, Christopher (2020) Medicinal Mushrooms - The Essential Guide. StoreyPublishing. USA.272p.

THIBAULT, Maurice and TWEDEEL, Russell J. (2016) Mushrooms: Bioactive molecules of medical and pharmacological interest. Multi-world editions. CANADA. 577p.

Yuan, Y., Wang, Y., Sun, G., Wang, Y., Cao, L., Shen, Y., Yuan, B., Han, D., & Huang, L. (2018). Archaeological evidence suggests earlier use of Ganoderma in Neolithic China. Kexue Tongbao/Chinese Science Bulletin , 63 (13), 1180–1188. https://doi.org/10.1360/N972018-00188

Özkırım, A., & Küçüközmen, B. (2022). Chitosan-based gel application on model bees ( Apis mellifera L.) for healing bite wounds caused by Varroa destructor . Journal of Apicultural Research , 61 (1), 45–51. https://doi.org/10.1080/00218839.2021.1935129

Stamets, PE, Naeger, NL, Evans, JD, Han, JO, Hopkins, BK, Lopez, D., Moershel, HM, Nally, R., Sumerlin, D., Taylor, AW, Carris, LM, & Sheppard, W.S. (2018). Extracts of Polypore Mushroom Mycelia Reduce Viruses in Honey Bees. Scientific Reports , 8 (1). https://doi.org/10.1038/s41598-018-32194-8

von Arx, M., Moore, A., Davidowitz, G., & Arnold, AE (2019). Diversity and distribution of microbial communities in floral nectar of two night-blooming plants of the Sonoran Desert. PLoS ONE , 14 (12). https://doi.org/10.1371/journal.pone.0225309

Frizzera, D., del Fabbro, S., Ortis, G., Zanni, V., Bortolomeazzi, R., Nazzi, F., & Annoscia, D. (2020). Possible side effects of sugar supplementary nutrition on honey bee health. Apidology , 51 (4), 594–608. https://doi.org/10.1007/s13592-020-00745-6

Tawfik, AI, Ahmed, ZH, Abdel-Rahman, MF, & Moustafa, AM (2020). Influence of winter feeding on colony development and the antioxidant system of the honey bee, Apis mellifera. Journal of Apicultural Research , 59 (5), 752–763. https://doi.org/10.1080/00218839.2020.1752456

Oskay, D. (2021). Effects of diet composition on consumption, live body weight and life span of worker honey bees (Apis mellifera L.). Applied Ecology and Environmental Research , 19 (6), 4421–4430. https://doi.org/10.15666/aeer/1906_44214430

Barker, R.J., & Lehner, Y. (n.d.). AND SUCROSE SYRUP AS MAINTENANCE FOOD FOR CAGED HONEY BEES .

Olaitan, P.B., Adeleke, OE, & OOla, I. (2007). Honey: a reservoir for microorganisms and an inhibitory agent for microbes. In African Health Sciences (Vol. 7, Issue 3).

Agarwal, R., Bansal, A., Saini, A.S., Raj, A., Kumar, A., Saini, S., & Gharde, SK (2023). Bee nutrition and artificial food. ~ 1635 ~ The Pharma Innovation Journal , 12 (6), 1635–1641. www.thepharmajournal.com

Alburaki, M., Madella, S., Vu, P., & Corona, M. (2022). Influence of honey bee seasonal phenotype and emerging conditions on dietary behavior and susceptibility to imidacloprid. Apidology , 53 (1). https://doi.org/10.1007/s13592-022-00922-9