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A data driven response to “Aspergillus in Cannabis: Evaluating the Evidence and Crafting Sensible Policies”.

Kevin McKernan, Sherman Hom, Ben Amirault, Yvonne Helbert, Steve McLaughlin, Liam Kane, Brendan McKernan, Matthew Brown, Nathan Houde, Heather Ebling, Victor Magaletta
Medicinal Genomics, Beverly, MA 01915

On March 31, 2025, the Society for Cannabis Clinicians (SCC) posted an article on the evidence for Aspergillus pathogen testing (A. flavus, A. fumigatus, A. niger, and A. terreus) in Cannabis authored by members of the Cannabis Industry Alliance of Oregon (CIAO). While we agree with these authors that the cannabis microbial testing standards need considerable reform, we believe organizations in the testing community have a unique perspective on this problem. It is not uncommon for organizations like the CIAO (representing producers) to argue for reduced testing. We offer this data with acknowledgement of our own conflicts, since we are also vested in the cannabis testing market. However, we believe the best resolution to these opposing conflicts is data transparency.

Collecting Cannabis Microbial Testing Data
In this spirit, we have collected cannabis microbial testing data from 14 states through Freedom of Information Act (FOIA) requests organized by Yasha Kahn of MCR Laboratories (Kahn et al.). We independently FOIA’d the state of Massachusetts to peer review Kahn et. al’s work and received identical data, albeit with different anonymization of the METRC barcodes the state used to ensure laboratory confidentiality. We then constructed a public web-based viewer of these testing failure rates at https://medicinalgenomics.com/lab-testing-data-portal/.

What is evident from these data is that the vast majority of commercially-grown cannabis passes required microbial testing. In fact, the failure rate for Aspergillus pathogen testing in states like Michigan and Nevada, which have been testing for the pathogen the longest and with the most transparency, is just 2.6-3.8% (Figure 1).

Additional data collected by Kahn et al, shows that the failure rate for all microbial testing in California is below 1% and dropping (these data are PDF collections and do not itemize individual microbial tests)(Figure 2). California only uses species specific testing for 4 pathogenic Aspergillus, E.coli and Salmonella.

The authors from the CIAO, when discussing the ubiquity of environmental Aspergillus, are confusing all 400 known species in the Aspergillus genus with the 4 pathogenic
species that are tested for in Cannabis (Wang et al.). There is no point in this conjecture when millions of tests have been run in the industry and public data exists as to its actual prevalence in various states and throughout seasons.

Calculating the Cost of Aspergillus Testing

The CIAO article suggests a $5 billion Aspergillus testing tax on the cannabis industry, which includes:

• Lab testing costs
• Remediation of microbial contaminants prior to sale
• Cleaning of facility
• Preventative maintenance and modifications of facilities
• Waste

However, the document later recommends that growers implement various pre- and post-harvest practices to reduce their microbial risks. As a result, it doesn’t seem fair to include the cost of cleaning and preventative maintenance modifications in the calculation, since the authors are recommending those actions. We agree with many of those recommendations as Aspergillus is not the only pathogen tested for and those requirements will not be relaxed if you simply remove Aspergillus testing. Hence, Aspergillus pathogen testing should not bear the cost associated with activities required to pass other regulations.

Furthermore, the assumptions made about the size of the cannabis testing market do not align with recent data. For example, the amount of waste generated from Aspergillus failures is significantly less than the authors assert. The Cannabis industry is estimated to be a $34 billion industry in 2025 (Whitney economics). A 1% failure rate would place this cost at $340 million if one assumes zero remediation or extraction of failed samples, which is allowed in many states. This also assumes the entire $34B Cannabis market is tested raw material, where a considerable portion of that market is packaging and non-raw material cost of goods sold.

A second market report places the entire Cannabis lab testing market in the US under $829M per year. This includes pesticides, heavy metals, cannabinoids, water activity and microbial testing. The Aspergillus pathogen test is just 1 of the 3-7 microbial tests required by at least one US jurisdiction. All of the Aspergillus tests in these 14 states reach a maximum of 311,095 tests/year in 2022 as seen in Figure 3. Even if you were to attribute the entire cost of all microbes in a microbial screen (labs often run 3-6 tests for E.coli Sal, TYM, BTGN, Coliform and TAC) to Aspergillus at $100/test, this amounts to only $31M in testing costs per year.

A better way to assess the market impact was published by Valdes-Donoso et al. where you take the 50 pound batch size in California ($25,000 @$500/lb) and apply the $50-$100 test price. A mere 0.2%-0.4% of the cost of the batch, not the 14.7% of the entire cannabis market ($5B/$34B) suggested in the CIAO article. In California, over 99% of the samples pass this test. Multiple market reports and public data refute the $5B CIAO estimated cost of Aspergillus pathogen testing.

The likely cause for this discrepancy is CIAO’s use of Oregon as a proxy for the rest of the country. The low failure rates detected in states like California, Nevada, Michigan, Hawaii, and Alaska may come as a surprise given that Oregon’s Aspergillus testing failure rate reached 11% with just 5,601 tests (Figure 4). Oregon is an outlier for Aspergillus pathogen testing. The labs had to implement testing under tight timelines prior to proficiency testing (PT) to certify any laboratories. As a result, there were some labs with failure rates as high as 64%. If you allow for any lab in the state that produced a single Aspergillus test, the variance in the lab to lab results compared to Nevada and Michigan is striking (Figure 5). This testing was run for only 4 months before legal pressure was applied to the regulators for test removal.

It is important to note that the regulators are tax funded and cultivators produce more tax receipts than testing labs while the consumers have little financial voice in this matter.

Double Standard
The SCC article suggests removing Aspergillus pathogen testing requirements and retrospectively collecting more data regarding causality. This approach fails to address the elephant in the room regarding the challenges in collecting such safety data with federal prohibition. The reason the Benedict et al. study was even permissible at a federal agency like the CDC is that it relied on Insurance claim data with actuarial consequences. Any attempt to derive perfect causality would require controlled studies that are not permissible under federal prohibition.
The authors are also missing the testing ethos that exists in regulating food that has carried over into the cannabis industry.

If the ‘perfect causality of death’ metric was applied universally, we would not be testing for pesticides, heavy metals or even cannabinoids as none of those have been linked to an acute death in Cannabis where Aspergillus has. If we are not willing to test for the only documented causative agents for loss of life in cannabis users, why test for anything?
There is literature on these non-microbial analytes being harmful in food or other settings, but it’s usually a long chronic exposure that leads to a very difficult causality test. The Aspergillus pathogen is the only analyte that has been documented in the cannabis literature as an acute cause of death via Aspergillosis (Gargani et al., Hamadeh et al.,Faccioli et al.). Non lethal cases are also documented in Zaga et al., Remington et al., Kaufman et al., and Marks et al.. All four of these cases required long and expensive treatments on immunocompetent patients. Anbar et al. describe an immunocompetent cannabis patient requiring lobectomy from Aspergillus niger. Many more of these case studies are documented by McKernan et al.
The CIAO article also made some assumptions regarding the Benedict study suggesting the ICD10 codes speak purely to immunocompromised patients. However, this is not true because 57% of the cases were immunocompetent and this is also reflected in the literature above.

Such invasive clinical work ups are not documented for pesticide and heavy metals found in cannabis. This is not to disparage the necessity of pesticide and heavy metal testing. We are advocates for pesticide and heavy metal testing as many of those analytes are neurotoxins that contaminate the cannabis-schizophrenia associations that are often used to push for prohibition. The schizophrenia field has also suffered from prohibition in that most studies never chemotype the cannabis. With CBD being studied in clinical trials to treat psychosis, this results in extraordinary confounders to these studies. Nevertheless, schizophrenia was listed as the main reason why Florida Governor Ron DeSantis rejected adult use cannabis reform in the state.
Many commonly used fungicides like myclobutanil, produce hydrogen cyanide when vaped and are enriched in vape pens due to the selective concentration of these analytes during extraction (Russo et al., Russo et al.). Many others are endocrine disruptors or neurotoxins known to induce psychosis (Serrano-Mendino et al.). We will never get to the bottom of this crucial debate if we remove pesticide testing, because we can’t find immediate death after consumption of pesticide contaminated cannabis. Their risks are superimposed from other industries.

Pesticide testing works. A comprehensive study in Canada surveying 327 pesticides, showed a 92% positivity for pesticides in the black market and only 6% in the tested markets (Gagnon et al.). You will also note that as California removed Total Yeast and Mold Testing and implemented more focused Aspergillus testing, pesticide use went down (Figure 2).

Causality is matrix independent
It is important to emphasize that our food testing standards do not rely on proof of E. coli that is found in lettuce not having enough deaths from lettuce alone to be a concern. We know Shiga Toxin-producing E. coli (STEC) has created illness and deaths from red meat, and if it is found on a new matrix, its pathogenicity does not need to be re-established on leafy greens or cannabis.

Fungal infections are estimated in 2024 to cost over $11.5B annually in the US with Aspergillus being the most frequent infection (CDC). Aspergillosis alone is a $1.3 Billion cost to the health care system in the US as of 2019 and is rising with its increasing presence in COVID-19 related fatalities (Garcia et al.). Both cannabinoids and nicotine are published with therapeutic activity against COVID-19 suggesting introduction through cannabis use described in the Benedict study is a valid concern (Breemen et al., Tizabi et al.). The CDC review on fungal infections notes:

“These numbers are based on administrative coding data. Due to underdiagnosis and underreporting, these data likely underestimate the true burden of disease.”
This underscores the precedent in public health to prevent harm through species-specific testing, even when deaths are not widespread in a given matrix — much like how we regulate E. coli in lettuce or Listeria in cheese.

Digging in the wrong place
What is also apparent in the Kahn et al. data release is that Total Count tests like Total Yeast and Mold (TYM), Total Aerobic Count (TAC), Total Bile Tolerant Gram-Negative bacteria (BTGN), and Total Coliform (Coliform) tests produce an order of magnitude higher cannabis failure rate than species specific testing like E.coli, Salmonella and Aspergillus (Figure 6).

 

The Total Count Tests are a de facto ban on beneficial microbes which are the best defense against these pathogenic microbes. Non pathogenic Trichoderma and Bacillus have been shown to push pathogenic Aspergillus out of the niche and these biocontrol microorganisms trigger the Total Count tests (Calistru et al., Octarya et al.) .

This de facto ban on beneficial microbes leads to a cat and mouse game of using multiple low concentration pesticides so no one pesticide is over the limit. Even worse, novel pesticides get used to evade the pesticide testing menu and have been detected in multiple studies (Sullivan et al., Gagnon et al., Russo et al.). Pesticide testing isn’t a cumulative test. You can have 10 pesticides all under the limit but their sum is not considered a risk.

The second issue one needs to consider is that there are no adequate standards for Total Count Tests as you can’t make 1,000-1,000,000 microbes and prove your test hits them all. So, instead, the TYM tests are Proficiency Tested (PTed) with a focus on just 2 organisms for TYM testing (Candida albicans & Aspergillus flavus). Candida albicans is not known to colonize Cannabis. Using a whole genome shotgun sequencing of over 1244 cannabis cultivars, Kannapedia.net has the most comprehensive public documentation of the cannabis microbiome and Candida albicans has yet to be found over 2 reads/million in cannabis flower underscoring the inadequacy of current Total count test standards.

Contrast this to species specific testing where the standards are readily available from multiple providers as the target analyte is precisely defined and these tests target microbes that have in fact been found in Cannabis. Why have a Total Count test that impacts the growers’ yield if you can’t make legitimate standards for such a test? This is the source of much lab shopping frustration in the cannabis market. Highly variable total count tests with no standards afford lab shopping.

In the spirit of transparency, we have also publicly listed the heterogeneity of the microbial testing regulations by state, demonstrating a growing consensus on relaxing the use of total count testing with adoption of more standardized tests.

For example, over the last six years, the fraction of US jurisdictions with legal cannabis programs that require a total aerobic bacterial/microbial or a total bile tolerant Gram Negative Bacterial count test has decreased by 17% and 29%, respectively. On the other hand, the fraction of US jurisdictions that require an Aspergillus pathogen and/or a STEC test has increased by 34% and 26%, respectively.

Some states are lifting the TYM limits to 100K CFU/g (Michigan, Connecticut, Maryland, and Florida), or making them a non pass/fail test (an indicator test as in New York) or eliminating them all together, e.g., California, Hawaii and New Mexico. We have been actively advocating for No pass-fail criteria (indicator tests) or Total count test elimination so the testing labs can focus on pathogens we have standards for.

Most importantly, Total count tests cannot satisfy Koch’s postulates, which is required for the Bradford Hill tenets of causality. Koch’s postulate requires that one isolate a single pathogen and reintroduce it to a host to demonstrate its pathogenicity. This cannot be performed on Total count tests as one does not know which microbes one is counting. As a result of the failure of specificity for total count tests, one cannot find clinical literature discussing 10,000 CFU/g being safe and 11,000 CFU/g being harmful as no causality can ever be determined. As a result of this, all of the clinical literature is written in the language of species specific tests.

The CIAO study also assumes that Aspergillus is an epiphyte and thus an unavoidable ubiquitous microbe. This is not true. Aspergillus is an endophyte and an epiphyte and can gain access to the plant through the substrate it is grown in via the root system. Testing substrate for Aspergillus is highly recommended.

These Aspergillus ubiquity arguments are not rooted in actual prevalence data observed across millions of cannabis tests and conflate all 400 Aspergillus species with the 4 human pathogenic species that are actually tested for. Much of the confusion over Aspergillus ubiquity lies in the testing volumes sampled. The studies cited to make these claims of Aspergillus ubiquity use air sampling of cubic meters to count all 400 species (Curtis et al.). Cannabis testing relies on 1g or 1ml sampling. This is a 1 million
times smaller volume being sampled in cannabis testing with a 100 fold exaggeration of the Aspergillus genus with the 4 Aspergillus species being tested in cannabis. This 100 Million fold exaggeration should be abandoned given the large volume of public testing data on Cannabis Aspergillus prevalence now in hand.

Punja et al. has also made note that the microbial burden on cannabis is correlated with the cultivar being grown, suggestive of heritable traits that can consistently alter and predict the CFU/g by orders of magnitude. This implies cannabis genetics and substrate cleanliness are critical aspects of successful growing strategies. The low fail rates in many states imply this is a manageable problem with proper strategy and nullifies any claims of futility by ubiquity.

In conclusion, we would recommend a data driven approach to regulations that looks at multiple states actual Aspergillus pathogen failure rates compared to the vastly higher failure rates seen in Total count tests. We recommend species-specific testing as only these tests can satisfy Koch’s postulates. Only these organisms decorate the clinical literature as a result of this specificity. Only these tests can offer valid standards.

Aspergillus flavus, fumigatus, niger, and terreus, are known to be valid clinical pathogens with substantial economic burden. Their pathogenicity cannot be conveniently ignored because controlled cannabis studies are impossible to perform under federal prohibition. This is not the safety ethos employed in the food industry. Salmonella is a quantitative concern whether it comes from chicken or leafy greens. There is no higher body count in the cannabis literature than Aspergillosis and other pesticides and heavy metal tests performed in food and cannabis are not held to this double standard, where causality of immediate death in the prohibited matrix must first be established.

We have been making these recommendations to regulators for over a decade and they would result in 4 of our tests being removed from the market in favor of 1 single Aspergillus test. This would result in a significantly more cost-effective microbial screening process for manufacturers and consumers. This contradicts any accusations of our conflicts of interest driving this data driven approach.

It is also prudent for the voice of the consumer to be heard in this discussion as they do not have the organized lobbying power of the large manufacturers on the Board of the CIAO. In some states like Connecticut, Hawaii, Michigan, Nevada, and New Mexico, where Total count tests have been relaxed, they have implemented species specific testing to address consumer complaints. If Total count tests were completely removed and generated significant consumer concern, they could be replaced with more targeted, lower failure rate tests for other specific microbes that have literature support for human pathogenicity such as Fusarium and fusariosis or Fusarium derived mycotoxins or mucormycosis. This focused strategy results in lower failure rates and laser-like specificity on quantifiable clinical risks that can survive legal scrutiny in a post Chevron doctrine regulatory environment. High fail rate tests without standards are unlikely to prevail under such scrutiny.

In summary, we agree with the CIAO/manufacturers that the cannabis microbial testing is in need of reform, but we ask that the community look at the data and target the tests that resemble tarot cards, lack standards, and induce the highest failure rate and lab shopping in the field. A close review of these data leads to an obvious conclusion. Relax or eliminate the Total Count tests and use species specific tests for lower failure rate and more clinical relevance.

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