Episode Transcript

Trevor:  Kirk, we're back.

Kirk:  How are you? And how excited are you talking about mold?

Trevor: So excited. And yes, I made you get up early to talk about mold. It is literal minus 30 here today. So what else would we be doing?

Kirk:  There's no more growing outside.

Trevor: Very very little, very slowly. Maybe spores though when we talk a little bit about spores. So I am Trevor Shewfelt, I'm the pharmacist, and you are...

Kirk:  I'm Kirk Nyquist, I am the registered nurse.

Trevor: And together we are Reefer Medness. So I'm excited about this one.

Kirk:  I know you are, this is a very geeky one.

Trevor: Very geeky.

Kirk:  I'm going to give the audience a head up, we're going to be talking about science and mold.

Trevor: Science and mold, and irradiation.

Kirk:  And irradiate.

Trevor: So I'll do my, I think I'd tell this, the cookie story. Every time irradiations comes up, I'm going to do the cookie's story one more time. So I grew up next to a nuclear reactor, a nuclear research facility. And one of my hockey coaches in high school, one of his sort of areas was irradiation. So they were irradiating. Things like carbon fiber to make them stronger for things like airplane parts and stuff. But as sort of a public relations thing, they irradiated some flour, made it into cookies and gave out the irradiate cookies at public events. I have eaten several irradiating cookies and it didn't do anything to my hair at all.

Kirk:  No, except you glow at night sometimes.

Trevor: Something like that.

Kirk:  Your wife can read off of your glow.

Trevor: Something like... That. So, real quick, we got a couple people from McGill, while the paper was more, but Saji George and Mamta Rani were the two we've got on the file. They irradiated some cannabis to, well, they had some cannabis irradiate. They worked on the what is left over afterwards. So when it comes to a radiation is there to get rid of the bad other stuff growing in it. So mold or fungi, some mold or fungi produce toxins, so those are the mycotoxins that you hear about, plus some bacteria, plus maybe some other stuff, you know, cannabis is a natural product, things get on it, things grow, cannabis has grown and stored in humid conditions. So you have some of this mold. Bacteria, endotoxins, microtoxins on it. We'll talk a little bit afterwards how harmful that might be but we're mostly in this case thinking about medical use for like someone whose immune system isn't so good like an HIV patient, cancer patient undergoing chemo. So those are the kind of things to keep in mind but before but before I let our guests chat about what they did anything else you want to tell the audience on the way in?

Kirk:  Well, just where did you find this story, because it's truly, as you sent it to me, you pushed it to, I Googled it and I found it pretty much everywhere. This research is everywhere, like in the publications.

Trevor: Yeah, yeah, if you've been following our social media, we are trying to, because there's so many, so many stories out there, we're trying to just put some headlines into our social-media feeds, because A, we think they're interesting, B, we can't fit everything we find into the podcast, and C, frankly, we get some interesting reactions. And this is one that was in our social medial feed. Extra bonus for it being a Canadian study. But oh my goodness people got excited about irradiation and maybe we'll talk about that a little more afterwards

Kirk:  Sure, let's go into it. Let's listen to the interview.

Saji George PhD: Excellent. And thank you, Mr. Trevor. I really appreciate you calling us on for this podcast. Myself, Saji George, and I am an associate professor at the department of food science and agriculture chemistry. I'm also chair of the department at the moment. So this is from University of McGill, Quebec, Canada. We work on many aspects relating to you know, the safety of food and food-related compounds, and then came to us, since the cannabis was legalized, came to use to verify what is the safety of things that are going in cannabis domain, and that's how we started this work. Other than that, we also work with many of those finding solutions for these kind of challenges, making our food system, food supply chain, and other kind of producers safe and sustainable. So we work with bacteria that actually produces some of the bio-stimulants and bio-control agents, which actually protects the plant from these kinds of toxins or these kind of pathogenic microorganisms. That forms one of the major domain of our work. Apart from that, we also look into some of those materials that we consume that are getting into our diet, for example, nanomaterials. How are they, and what is the implication of those nanomaterials in food in specific disease conditions like industrial bowel disease, et cetera. So that forms into, to summarize this, majorly we are working on safety of food and also we're working a little bit on the sustainability part of agriculture. So Mamta, over to you for your introduction.

Mamta Rani: Myself Mamta Rani and I'm assisting Dr. Saji in all these projects he mentioned and we are mainly working on bio-stimulants, but this project what we will be discussing today was on mycotoxin of cannabis and we will we discussing like how we started, what we did and what are the outcome of this project. Dr. George will explain it more.

Trevor: Thank you. So just for the audience, I'll read the title and then we'll we'll talk about the paper. So "Detection of Mycotoxigenic Fungi and Residual Mycotoxins in Cannabis Buds Following Gamma Irradiation." So how about give us a little bit of a background? So why and you talk about a lot in the paper, but why does fungus on cannabis matter? Why would a cannabis consumer care?

Saji George PhD: Very important question. So as we know, nowadays, many food producers or these cannabis producers that are coming into the market, they are not sterile. And fungus actually has a major role because these guys can grow on diverse kind of matrices. And cannabis being grown under humid conditions, cannabis being the bud being very moist and kind of stuffs, they are particularly vulnerable to fungal attack. Fungal attack in the sense fungus spores, they can go and then they can sprout there and the fungus can grow. And obviously when the fungus grow, they actually produces the secondary metabolites called mycotoxins, which they produce for their own purpose to make them more competent in that matrix or in the microimmune where they are growing. But those secondary metabolite are extremely toxic to humans. And so, and I have to also mention that the fungus is not just getting from the growing or the place where the cannabis is grown, but down the lane for the food in the supply chain, they can enter at any node. It can be during the storage, it can be doing their transportation or, you know, at a retailer, for example. So at any stage, these kind of funguses can come in contact with these kinds of substance and when they grow, they can actually produce toxic. So there is this question about monitoring the cannabis producers for their sterility in terms and their suitability in terms of consumption. So that question, as you can see, has not been addressed too much. There are some publications in the domain in literature. Talking about the identification of microtoxins or these microtaxing producing fungous in cannabis producers. But then we asked that question, like the industry standard, the best practice for processing these produce in order to get rid of those kind of obnoxious pathogens or their metabolite, the best practice is to subject it to gamma radiation. There are many other ways of Play session. But gamma radiation is the number one in terms of its astringency or efficiency in removing the pathogens, in removing their spores, and also in removing the metabolite. So now we ask that question, is that really capable of removing it? Why I stress on that really part is... When you test in conventional, like if you take a cannabis produce that is coming into the market and subject it to the normal industry standard kind of testing protocol, you may see it's sterile. It's good. It's a good to go kind of. But we ask this question, first, is gamma radiation good enough to completely remove the sports and the mycotoxin and also the reminiscence of these DNA fragments? That are that in a part of the genes that actually are in the formation of this mycotoxin. And also we asked this question, current testing methods, are they good enough to test in order to pick up the reminiscence of these omniscient agents present on the cannabis products?

Trevor: Okay, and you did mention in the paper, so I'll just highlight for people, you know, I'm an otherwise healthy adult, so you know me inhaling a fungal mycotoxin might be good, might be bad, might affect me, might not, but it seems to be an especially big deal in the medical end of cannabis if we have someone who's immunosuppressed, a cancer patient undergoing chemotherapy, an HIV patient. Other people who, for whatever reason, their immune system isn't up to snuff, inhaling bacteria, fungus, mycotoxins might be especially bad for them. So it seems to be an especially big deal for the medicinal end of things.

Saji George PhD: Exactly. So I was coming to that point. So what with these kind of things present in these producers, like if it is a very low quantity, whatever it is, if it is passing the industry standard, why should we bother about this? That's the question, right? So, you know, if you know if it's a normal healthy person, as you mentioned, may not be much of an issue. Obviously there is issue, but it may not manifest immediately. Like, you know, it may add on to the smoking, for example, it may add onto the burden to the lung, like the toxic impact or the implications on the lung. But this issue becomes a very evident and immediately hazardous for people with these kinds of medical conditions, where their immune system is not up to the mark. When are the patients or people who are taking it for medicinal purpose usually may be undergoing a palliative therapy, pain management, or maybe for some other reasons, people may be low-wage immunate. People may have. Some issues with the immunity. And that's where even a single spore actually matters. If the thing that goes into the lung, then there is now protection provided by the body through the immune system. As a result, it can actually manifest as an infection of the lung or the mycotoxin. The smallest quantity that is present can actually cause injury to the lung tissue. yes.

Mamta Rani: Can I add something?

Trevor: Absolutely. Please.

Mamta Rani: Yeah, because in the US, there are more than 19 cases reported so far of the same like when people were consuming microtoxin in the cannabis and there were so many fatalities. But somehow, I don't know, I did not see many reports in Canada since cannabis got legalized. Only few cases where it was just this reaction, like no fatality, no complication, just a little bit like some adverse reaction they call it. So I don't know whether our doctors are very careful in prescribing or our producers are making it, producing it very carefully. But in the US, I see there were. 19 cases reported in last, more are lasting 10-15 years. There were some cases in 1988 and 1989 case and more are after 2010. So I am really, there is a report on cannabis in Health Canada, But not many reports I'm really surprised. Is it because of the regulation? It's the same regulation. I am really concerned about the regulation because mycotoxin in cannabis is treated same as in the food testing industry. It's same because it should be actually different. That's our biggest concern here. It shouldn't be treated as other food commodities. It should be treated special actually.

Trevor: Okay, no, that's a good, good point. I'm glad that we don't have a lot of key supports in Canada either. And I'd love to think it's because our growers in Canada are especially careful. But yeah, I can see why it would be, it should be different for something we inhale into our lungs than something that goes through our stomach. That makes a lot of sense.

Saji George PhD: Maybe I can add on to that why it is lesser in Canada. I mean it was very interesting. I was just reading about the Health Canada report where they actually looked into these kinds of pesticides, mycotoxins, heavy metals in legal and illegal sources. The cannabis that is you know sourced from legal and legal sources and they show that when it is coming from illegal sources almost all the samples they are actually having these kind of presence of mycotocins or pesticides or heavy metals, all those are likely to cause injuries to the lung or through the burst. So I believe down the border, I'm not sure how far is the legal producers getting into the market, or it may not be that stringency in the production, and the testing is as stringent as a canary. So I just wanted to add on that part. Now once we've legalized it, it becomes a little bit better in terms of the product quality. I just wanted to add one more point. Like we discussed why are we bothered about the presence of mycotoxin in this product from a person's medical point of view, something like if there is a compromised immune system, it may be more deleterious. But also this is a workplace safety concern. Like there are people who are working in these kinds of industries. It is actually a closed kind of a place where there is... High humidity, high presence of these, and there is lots of these. If you take the particles from these facilities, there are likely to be presence of the spores or the mycotoxins or the fungus spores are likely be present. So it may also show that there is a chance of an implication for workplace safety as well. I mean, it's not directly related to the work we did. It implies that if there is a presence of these funguses or spores in the produce, obviously we can imagine that could also be present in the in the atmosphere where it is grown.

Trevor: That's a good good point. Go ahead.

Mamta Rani: To preserve the potency of the cannabis buds, it has to be processed under certain temperature and humidity has to. So in that environment, it's very good environment, like perfect for fungi to harbor. So if there is even small spores there, they can, you know, in that humid and high temperature, it can really flourish. Big concern, actually, for the workers who is handling and who is working. For their health is a huge concern.

Trevor: Okay, so now let's move on to the actual paper. So we've got the problem sort of laid out. What did you guys do and then what did you find?

Saji George PhD: So I just mentioned that industry practice for sterilizing this is through the different strategies. But one is the gamma radiation. It's a very good method. So first, we worked with an industry partner where they supplied us the materials which is not going through that gamma radiation and the materials that is going through the gamma radiation to see the effectiveness. We tested these materials for the presence of the fungus all together. So how many, what type of fungus are present. So we have done culturing them to visualize them. And also we have something that we cannot see. We can actually do go through another tool called metagenomics. So we take extract all the DNA and then we isolate and then run through those kinds of instruments which can tell us what kind of a fungus or varieties of funguses are present, and also bacteria that are present. So we do those kinds of metagenomics or ITS, all these to actually identify the microbiome generally to say, or type of bacteria and fungus present in the sample. What is our finding? The finding is gamma radiation is really good in reducing those pathogens. In fact, most of those of nauseous or pathogenic funguses and bacteria, they will be taken care of, which means if you are doing the proper way of gamma radiation, it will actually kill or eradicate most of those fungusses and bacteria. But to our surprise, what we find is it is not completely sterilizing. Gamma radiation is supposed to be a sterilizing agent, but it's a radiation. So obviously there may be shadow regions where this extremely difficult kind of form, especially the spores of the fungus can still stay and it is not completely eradicating or eliminating. In a sense to say that it is a total sterilization protocol. So that's what I was defining from that part. And then we also. Extended these examination into samples that we collected from the legal sources. So, you know, we sampled from SQD and SQ.

Mamta Rani: It's good to see. Actually, it was unknown samples. We cannot...

Saji George PhD: So we don't know who produces it, but we are actually.

Mamta Rani: Blindfolded.

Saji George PhD: blindfold. We are taking from legal sources. And we conducted the same kind of assays. And there we also, I mean, I have to also tell you that we are not really restricted to the ELISA-based assay. We also went on to mass HPLC-based analysis in order to see the compound beyond what we can actually be detected by the traditional assays So So when we do that. What we identify is, in traditional assays, you may find it absent, which means in the industry standard wise, these are the standard protocols that are used. It is below the detection limit of these traditional assay that are use by the industry. But if you are going a little bit more, like advanced instruments, then you can pick up the signals of mycotoxins. Even those are present in the samples that are coming into the market, although most of the sample, I should say, they are pretty good in terms of the fungal load or in terms of the mycotoxin. But if you are actually doing with advanced instrumentation, to pick up the toxins as the molecules or to look for the genes that is corresponding to the toxin. If you are using PCR-based techniques, so we will be able to find the presence of these things in many of those samples that came to the market. Especially one sample was really bad, right? I mean, some of them. So in terms of, you know, this is also coming to the legal sources. I mean illegal in the sense is where we can, you know it's presumed to be a good high quality. Showing that there are cracks in the system, in the quality control of these materials that are getting into the market. You want to add something?

Mamta Rani: Actually, big concern was the viable spores, because viable spore, they cannot be detected with even HPLC or LC-MS, these advanced instruments. They will be detected only with very traditional culturing techniques because these scores cannot be detected. And they can even tolerate these gamma radiation or heat sterilization very easily because they have very thick spore wall that can protect them from heat and radiation. And these spore, even a single spore, it can be inhaled by any immunocompromised person and can colonize their lungs will be very, very fatal. So, presence of spores was big concern and I found one sample was having spores of aspergillus flavors that produce aflatoxin. There were some other like minor, but this was the big concern. Aflatoxin producing hungous spore presence was detected in one of those samples.

Trevor: No, and that's fascinating. And we won't, people can read the paper if they want to see, you know, that you did, you know culturing and then you did QPCR and then you did Eliza and like you said, you did some HPLC. We'll leave some of the details, but, so you did find that if you use basically more stringent testing, you're finding, you know, not in everything, not all, but we're fine where it would have normally passed. We'll call it Health Canada standards, that there actually still are some harmful both spores and mycotoxins in there. So now that you've found that, did you put any thought into what the industry should do to, you know, what should we do about that?

Saji George PhD: Very good question. Before answering that question, let me first tell you, when you find a mycotoxin that is present, which is not in according to the normal industry standard, this produces or this product may be good to go. But you have to understand there is no tolerable limit for mycotocin. Even, you know, usually the, we say 5 PPB, that So I think, you know, according to the Health Canada standards. We put some regulatory limit for the presence of the mycotoxin that can be allowed in the products. You have to understand that that is mostly related to our ability to detect, not to say that one molecule of mycotaxin can be tolerated. No, it is the body. I mean, there is no tolerable limit for mycotoxin. I just want to say. So. Our finding that using advanced instruments we still can pick up the signal is significant when it comes to even normal human being and obviously there is more concern when it is a immunocompromised persion. Now coming to the next question, what can we do about it? There are a couple of things. So this is once we identify the, you know, this is about the risk mitigation. How can we reduce the risk of mycotoxins and fungus present in cannabis? First of all, it shows that industry has to have some stringent control over the production. So it shows, that obviously, I mean, like I alluded to you before, that when Health Canada's report shows that illegal sources contain more of these things than the legal sources, which means when it is legally procured, there are some industry standards, good agriculture practice that is going in, which actually helps in reducing these mycotoxins. So, obviously, that is the first thing we have to improve the good agriculture practices or practices that are fully in place in production stage itself. So the growers should be mindful about it and so that we can reduce it. So we have implement those kind of antimicrobial basically to take out the fungus from the facility. So whatever steps that we do or controls that we should be exercised. It also brings to the point that as regulatory stage, we have to implement stringent testing criteria, like looking for whether it is truly devoid of these kinds of fungus, the live strain, live spores, and the mycotoxin presence. So that means we have to go beyond some of those right now, the standards that are put in place is not good enough to detect. So probably you have to make more stringent kind of analysis of these samples. Basically, it can be an HPLC-based method or a MS-based where we can actually pick up the sickness of these chemicals. Now, having said these two mitigation things, I have to say that still it will be very difficult to actually get rid of, first of all, getting rid of a mycotoxin or a fungus that is established on a product. I told you gamma radiation is the best in practice, that's the best industry practice to actually stabilize the product. But the thing is, so that tells us that we have to prevent the generational mycotoxin at the very early stage and also throughout the supply chain. So one of the things that we are working on is to help the producers, the growers, to actually produce quality produce which does not have or which will minimize the chance of fungus establishing on plant. So that is through biocontrol. We have identified bacterial strains, for example, that will prevent these flat toxin producing or don't produce, is another mycotoxin producing fungus from establishing the plant on to the plant. So they produce some chemical compounds, these are like a bio-stimulant, I'm sorry, bio-surfactant that actually targets these kinds of fungus and then they can actually win the fungus from establishing on the plant. So at that production stage, we can actually help the plant to grow faster, better, because it's also a bio stimulant property and also control the fungus from establishing onto the plant, if they are not establishing on to the plant they're not going to produce the mycotoxin. So that is the idea. So we are working with the growers to actually take and our preliminary research show that it is really good in terms of reduction in the fungus itself. So even before the gamma radiation, we can see there's a substantial reduction in the number of fungus colonies that we can count. And also we are now testing on the microtoxin in these kinds of samples and we find it is substantially reduced in comparison to the control. So these are some of the strategies that we can implement in order to reduce the burden or the risk from mycotoxin or the fungus that causes this kind of infection in humans.

Mamta Rani: So as Dr. Saji mentioned, use of biocontrols, it will be actually taking charge of two things at a time. First, we can control mycotoxin and enhance the growth. At the same time, we can reduce the use of chemicals because another problem in cannabis is the presence of pesticides, fungicides, this is big because they are using it a lot to prevent. So by using these biocontrols, we can actually take control of those things also. So it will be really a good idea to promote these biocrontrol in cannabis industry because this way we can control pesticides and other chemicals or even because whenever we use any chemical pesticides or even fertilizers, they will always bring heavy metals and other toxic substance with it. So by using these, everything will be, you know, controlled. Everything will, it will help in all other aspects.

Rene:  Fascinating stuff. But that's as far as we're going to be able to go today. We'll continue this conversation about cannabis and irradiation and proof that Trevor is in fact a fun guy. We'll continued that next time on Reefer Medness - The  Podcast.