“You can just point to the fact that we have been deemed essential, why are we not legal?”
^ Matt Hawkins, managing partner of Entourage Capital, a private equity firm with $200 million invested in Green Thumb Industries and other cannabis producers, is hopeful that the COVID-19 pandemic will accelerate the cannabis legalization movement.
“Without providing these businesses the relief needed to carry out the recommended public health and worker-focused measures, we are putting these hard-working people—and ourselves—at risk.”
^ In a press release, Rep. Earl Blumenauer (D-OR), founder and co-chair of the Congressional Cannabis Caucus, advocated for cannabis businesses to have access to Small Business Administration (SBA) loan programs. He, along with Rep. Ed Perlmutter (D-CO), introduced legislation April 23 that would make cannabis businesses eligible for SBA COVID-19 relief programs. Source: U.S. Congressman Blumenauer’s office
“While there has been some progress in scaling back the war on people who use marijuana, it is still wreaking havoc in much of the U.S.”
^ In an April 20 report titled “A Tale of Two Countries: Racially Targeted Arrests in the Era of Marijuana Reform,” the American Civil Liberties Union (ACLU) found that racial disparity in cannabis arrests has increased since 2015, with black people more likely to be arrested for cannabis possession, even in states that have legalized cannabis. Source: ACLU
Cannabis sativa is the species comprising hemp (which is bred for fiber, seed and/or oil) and marijuana (which possesses tetrahydrocannabinol (THC)). Both varieties share many of the most common and serious diseases that can result in significant production loss.
Due to different growing environments (outdoor versus a controlled environment), hemp crops may have more exposure to disease agents than marijuana plants. Unfortunately, there is little pathology work done on Cannabis sativa crops, and many problems are not even diagnosed, let alone diagnosed properly. Below are diseases the Nevada Department of Agriculture (NDA) plant pathology laboratory has detected, as well as best practices for field scouting and disease management.
Diseases Caused by Fungi and Oomycetes
Fungi are the most common pathogens that infect many plant species. They produce mycelium (white filaments) and/or spores, and some produce special structures for winter survival and dissemination. Plants infected by fungal pathogens may exhibit symptoms of leaf spots, mildew, blight (more extensive browning or sudden death), wilt, canker or root rot, all of which can turn lethal. Oomycetes are fungus-like organisms such as Phytophthora and Pythium that commonly cause various diseases on plants, including on Cannabis sativa.
Fusarium wilt (Fusarium oxysporum and F. solani)
Fusarium wilt is a destructive disease on Cannabis sativa plants. The fungi, Fusarium oxysporum and/or F. solani, infect vascular tissue, causing plant wilt and death. When infection progresses systemically, pathogens can be passed from mother plants to cuttings. Infected plants can exhibit poor growth, may be unresponsive to added nutrients or fertilization, and can wilt even with adequate soil moisture.
- Diagnostic characteristics: Vascular tissue turning dark brown. A cross or longitudinal section of the basal or lower stem often reveals the vascular browning.
- Prevention/Treatment: The fungi survive in soil, growing media or dead plant tissue, and the disease may persist for years if not treated. Therefore, prevention is the best management practice for this disease. For marijuana crops, mother plants should be kept healthy and free from Fusarium infection through periodic monitoring and testing. (Most plant pathology labs at university extensions can do these tests. NDA’s plant pathology lab only accepts samples from Nevada.) Once mother plants, cuttings and/or growing media are found to be infected, they should be destroyed, and affected areas should be thoroughly disinfected with a bleach solution to prevent spread. For hemp crops, a field free of Fusarium diseases should be selected. If Fusarium wilt occurs in the first hemp crop, it is highly recommended to rotate with other Fusarium-resistant crops to mitigate disease pressure by starving the fungi.
Fusarium stem canker (Fusarium spp.)
Some Fusarium species cause stem canker or stem rot, in addition to vascular wilt. Two species, F. oxysporum and F. solani, are found to be associated with stem canker disease on hemp.
- Diagnostic characteristics: Affected plants exhibit dark lesions or cankers on stems, and white mold is often found on the surface of cankered areas. Internal tissue of stems may rot.
- Prevention/Treatment: Stem canker has more extensive infections on branches and non-vascular tissue, whereas vascular wilt is more restricted in vascular tissue, causing systemic wilt. Fusarium stem canker has not been observed on marijuana plants. Like the management practice for Fusarium wilt disease, the best way to avoid stem canker disease is to select a field that has not been cultivated with a Fusarium-susceptible crop (tomato, melon, sweet potato, etc.).
Fusarium root rot (Fusarium spp.)
Fusarium root rot is a destructive disease on hemp but is not seen in marijuana. Several Fusarium species, namely F. oxysporum, F. solani, F. redolens, F. tricinctum and F. equiseti, contribute to this disease.
Pythium crown and root rot (Pythium aphanidermatum)
Pythium aphanidermatum, a pathogen belonging to the oomycetes, attacks hemp crops, causing damping off in the seedling stage or death of plants during the season.
- Diagnostic characteristics: Early infection signs include mild leaf yellowing, curling or necrosis (death of plant cells) along the edge. Plants may wilt temporarily in warmer temperatures. Examining the crown area of the stem may reveal necrotic lesions and internal discoloration (visible by cross section). As the disease progresses, lesions or cankers will expand at the basal stem and infection may spread from the crown area upward to the main stem and lower branches, eventually killing the plant.
When scouting, look for water-soaked lesions and cankers around the basal stem when a plant starts to show foliar symptoms. A canker where the stem meets the soil line is the most diagnostic symptom for Pythium crown rot. During the late season, grayish to white-colored mold (Pythium mycelium) may be visible on the surface of the crown area. This pathogen can cause severe root rot in certain conditions; however, it appears to be more aggressive on crown and stem tissue, especially if the soil surface is covered by plastic mulch.
- Treatment: Removing mulch helps reduce soil moisture and disease incidence temporarily, but it may not stop the disease development in plants that have already been infected. A best practice to eliminate or reduce the disease is to treat seeds with approved fungicides or a diluted bleach solution.
Powdery mildew fungi are a group of widespread, well-known obligate fungal pathogens that negatively impact a diverse array of host plants in a variety of environmental conditions. Cannabis sativa plants grown indoors are prone to powdery mildew, and due to the difficulty of eradication and limited products approved for Cannabis, the disease can be troublesome to control. Specific fungal species causing powdery mildew on Cannabis are not fully known, but in one marijuana cultivation facility, the powdery mildew fungus was preliminarily found to be closely related to Golovinomyces ambrosiae. In Kentucky, Golovinomyces sp. was reported to cause powdery mildew on hemp. This is a significant finding because knowing the pathogen’s species will help develop solutions for effective treatment and management.
- Diagnostic characteristics: Powdery mildew is easily identified on plants: It produces patches of grayish white, powder-like fungal mycelium and spores on leaf surfaces. Fungal spores may be carried by air currents and disseminated inside a cultivation facility through air circulation.
- Treatment: Powdery mildew is considered treatable; however, treatment options for Cannabis may be limited. As such, once an infection takes hold, eradication of this fungus from the facility is recommended. Removing infected plants as soon as they are noticed and sanitizing the infected area with a diluted bleach solution or other disinfectants can help eradicate early infection and stop the disease from spreading.
Latent fungal infection in seeds
Cannabis seeds may carry some fungal pathogens with or without visible seed damage. Some fungi may infect Cannabis seeds, causing poor germination or affecting viability of seedlings. Some imported hemp seeds are found to be infected by species of Alternaria, a genus of fungi considered to be ubiquitous, opportunistic pathogens. Although they are frequently associated with hemp seeds, their impact on seeds and plant health is unknown. Another fungus frequently isolated from hemp seeds is Rhizopus, a fungus that causes fruit rot on other crops. Infection of hemp seeds by Rhizopus may impact seed germination and seedling health, but there is no experimental data to prove it.
- Diagnostic characteristics: Close examination of individual seeds may reveal some fungal mycelium growth during the routine germination test.
- Treatment: Treating seeds with a protective and approved fungicide or diluted bleach solution was found to improve seed germination and crop success. State extensions can also test seeds for pathogens.
Disease Caused by Bacteria
Bacteria are microscopic, single-celled organisms. Most bacteria are culturable on artificial nutrient media and cause soft rot (decomposed plant tissue), fire blight (“scorched” lesions), wilt or root rot.
There is a group of bacteria that is not culturable on nutrient media called fastidious prokaryotes. Phytoplasmas are one type of fastidious bacteria that live in phloem sieve tubes of live plants and are transmitted by insect vectors. Typical symptoms of fastidious prokaryotes include witches’ broom, systemic yellowing, stunting, chronic decline and abnormal growth.
Phytoplasma witches’ broom
Hemp is prone to phytoplasma infection, and once infected, plants exhibit severe witches’ broom-like symptoms.
Diseases Caused by Viruses
Viruses are submicroscopic particles made up of protein and nucleic acid. They infect plant cells and use the plant’s nucleic acid replication systems to produce more viruses to interfere with plant growth. Common symptoms caused by viruses are leaf mosaic, twisting, distortion, chlorosis, yellowing, necrosis, stunting or death.
Cannabis mosaic and leafroll
Leaf mosaic and other virus-like symptoms have been observed on both marijuana and hemp crops. These symptoms suggest viral infections, but no specific virus has been detected. Some cannabis growers consider tobacco mosaic virus (TMV) to be the culprit, but lab tests in Nevada were negative in several cases.
- Diagnostic characteristics: Leaf mosaic, twisting, distortion, chlorosis, yellowing, necrosis, stunting or death. Another virus-like symptom on Cannabis crop is leaf curling. Curled leaves appear yellowish or light green and feel coarse to the touch. It is unknown if this symptom is caused by a type of virus (such as potato leafroll) causing leafroll symptoms. Both mosaic and leafroll may occur on the same plant, suggesting dual infections. We have no formal report of leafroll virus in Cannabis.
- Prevention/Treatment: Viral diseases on Cannabis sativa are difficult to treat, and limited treatment products are available. Most viruses are transmitted by insect vectors, so monitoring and controlling insects is key to mitigating virus infections in the field. For indoor Cannabis cultivation, it is recommended to frequently wash hands with soap to prevent the introduction of TMV and other viruses.
By Shouhua Wang, Ph.D. Dr. Shouhua Wang is a plant pathologist at the Nevada Department of Agriculture and a lead scientist for the state plant pathology laboratory.
Study Shows A Link Between EVALI Cases and Adult-Use Dispensary Access
“One possible inference from our results is that the presence of legal markets for marijuana has helped mitigate or may be protective against EVALI.”
^ In a study published in the Journal of the American Medical Association (JAMA), Coady Wing, Ph.D., Ashley C. Bradford, MA, Aaron E. Carroll, M.D., MS, and Alex Hollingsworth, Ph.D., from Indiana University found that the number of e-cigarette or vaping associated lung injury (EVALI) cases was lower in states with legal adult-use cannabis dispensaries. “This association was not driven by state-level differences in e-cigarette use, and EVALI case rates were not associated with state-level prevalence of e-cigarette use,” researchers said. Source: JAMA
Democratic Senators Call for SBA Loan Access for Cannabis Industry
“We … ask the Subcommittee on Financial Services and General Government to include language in your forthcoming legislation to help extend SBA loan programs to legal cannabis businesses.”
^ Democratic Sens. Rosen, Wyden, Duckworth, Bennet, Gillibrand, Sanders, Markey, Booker, Menendez, Merkley and Harris signed a letter asking the Senate Appropriations Committee to include language in the Financial Services and General Government and Related Agencies Appropriations Bill (fiscal year 2021 ) prohibiting the Small Business Administration from denying loan applications to cannabis operations in state-legal markets. Source: U.S. Senate
COVID-19 Throws A Wrench in THE Cannabis Reform Movement
“People are scared. They don’t want to touch a pen or paper.”
^ Melissa Fults, executive director of Arkansans for Cannabis Reform, is one of many cannabis advocates who are seeing legalization efforts hit roadblocks amidst the global COVID-19 pandemic, as social distancing measures put ballot drives on hold and lawmakers grapple with the crisis. Source: Politico
“I don’t understand! How can this batch fail when previously tested cultivars all pass?”
“How can we have pesticides in our extract? We don’t spray pesticides on our plants!”
“How is there a presence of metals in our formulas? This doesn’t make any sense at all!”
It is unfortunate, but these are valid statements that have been made by many people working at cannabis operations. Contamination, potency reduction and other concerns can arise during the cultivation, processing and manufacturing of infused- cannabis products.
It’s no secret that problems arise in production environments; however, what delineates a successful cannabis producer from one in constant remediation is understanding analytical testing and its vital role in shedding light on the health and quality of your operation.
The statements above can be alarming to members of the leadership team who do not come from a scientific background. The cannabis industry has done a great job attracting talented people from diverse backgrounds in horticulture, consumer packaged goods and general manufacturing to bring a collective knowledge base to the table. Yet, when your industry is based on the foundation of biological and chemical principles, a member of the executive leadership team needs to be a scientist who can use his or her experience to discover the right analytical methods to monitor spikes in concentration levels of specific contaminants, propose solutions to mitigate harm to production and more. In addition, this person needs to help answer certain operational questions such as: Are my parameters sufficient for optimal extraction? Does quality remain when the extraction process is scaled up? Are we appropriately tracking all process data to analyze yields and run times to optimize production?
Century-Old Testing Methods (With Enhancements)
Testing laboratories in the cannabis industry use various analytical methods adopted from the environmental, pharmaceutical and petrochemical industries. The instrumentation used to quantify cannabinoids, identify contaminants and collect qualitative data is not new-found, either. Russian botanist Mikhail Tswett first developed chromatography techniques in 1903, using a column of calcium carbonate to produce a colorful separation of plant pigments.
Chromatography has since developed into an invaluable laboratory tool for the separation and identification of compounds. High-Performance Liquid Chromatography (HPLC) is the separative vehicle equipped with either an Ultra-Violet/Visible (UV/VIS) detector or Diode Array Detector (DAD) to properly quantify phytocannabinoids using their individual fluorescent values. The cannabis plant produces more than 150 known cannabinoids. Hence, without sophisticated instrumentation, analytical techniques and scientific competency, qualitatively separating the compounds and quantifying the ones of interest would not be possible. In cannabis potency analysis, 11 compounds are of interest, with cannabidiol (CBD) and tetrahydrocannabinol (?9-THC) being the most popular to current producers.
Data on potency across multiple harvests, cultivar types, growing techniques, etc., is not readily available; therefore, it would behoove cannabis producers to archive all potency analyses performed in-house and chart the data to get an accurate read on the business’s processes and performance. This would provide a database to refer to when making predictions on plant performance and system health, as well as give insight into the feedstock being used for further processing activities. You could easily predict the potency of your crude extract using this data archive and performing a quick calculation (e.g., percent of oil created from volume of material) based upon the extraction technology metrics (e.g., pressure, temperature and solvent flow rate). This arsenal of data also will serve you well in validating the third-party testing lab’s analytical results used to approve your product for sale.
Contamination in Cannabis
Contaminants are an ongoing problem that plagues industries working with agricultural feedstock, botanical extracts and metal machinery. In the cannabis industry, laboratories monitor the amounts of pesticides, heavy metals, micro biologicals and mycotoxins present in representative batch samples. These contaminants can be introduced into your processes in a multitude of ways and can be detected using various techniques and instrumentation.
Pesticides: Pesticides can be introduced at different stages of your operation, including in raw materials (the ingredients imported for infusions as well as biomass used for active pharmaceutical ingredient (API) extraction). Pesticide contamination of raw materials can occur at the third-party vendor’s facility due to pest management procedures or during transport of the materials to your facility. Many markets have lists of pesticides approved (by state agriculture or other departments) for use on cannabis, as well as allowable pesticide residue limits for products sold to consumers.
This does not pose much of a threat for product sold as retail flower; however, if the flower will be used as raw material for processing, potential concentration needs to be considered. Concentration in chemistry is defined as the “amount of solute per amount of solution or solvent,” according to the University of Texas. A solute is a solid, liquid or gas substance dissolved by a solvent to create a solution. The process of extracting cannabinoids from the plant material is, in chemical terms, a concentration step, regardless of the solvent used. If trace amounts of a pesticide are present in the flower that is transformed into oil for infusion, processors risk concentrating the contaminant beyond allowable limits.
Pesticides are generally quantified in cannabis plant material and extracts by using liquid chromatography with tandem mass spectrometry (LC-MS/MS) technology. Compound separation occurs using chromatographic techniques, and those compounds are then detected by a triple-quadrupole mass spectrometer, which allows for higher sensitivity to detect compounds at very low limits. This technique is commonly used to quantify pesticide concentration in various flower, extracted oils and infused products. When interpreting these data reports, it is important to pay attention to the actual concentrations of each analyte versus just a PASS/FAIL. Investigating further is essential to understanding the parameters, processes and efficiencies. You have to look deeper into the actual concentration levels of analytes/contaminants in your samples to notice any compounding trends leading you to a failed test batch in the near future, understand the concentrating effect of processing cannabis flower into oils and to ensure consistency in the process and product.
Heavy Metals: Heavy metals commonly screened in cannabis plant material and extracts are lead (Pb), cadmium (Cd), chromium (Cr), arsenic (As) and mercury (Hg) to name a few. These metals naturally occur in soils and rocks. They can also be found in aquatic systems from natural occurrences, like volcanic eruptions, and human activities, such as mining.
This means the soil source used to cultivate the plants and the water used for irrigation could already contain significant levels of metal. Applying the concentration principle, it is essential to monitor the soil and water closely to ensure there will be no issues in downstream processes. In addition to cannabis testing facilities, environmental laboratories can use several reliable methods to determine the levels of metal in sources. Metals are analyzed through Inductively Coupled Plasma and Mass Spectrometry (ICP-MS).
Microbials: Micro biological organisms (or microbials) can destroy crops during early cultivation stages. They can also lie dormant in the plant until conditions are just right and haunt you well beyond harvest. Microbial tests include total count tests that target large microbial groups such as: aerobic bacteria, yeast, mold, and coliforms (present in animal and human digestive tracts, as well as in plants and soil). Traditional culture-based plating methods are typically used to qualitatively screen for microbes. Screening can also be done using DNA-based methods. Mycotoxins (toxic secondary metabolites produced by fungal organisms that can cause disease and death in animals) can be detected using LC-MS/MS.
Typically, if there is a microbial issue in the plant material, further processing can be the answer to soothe some of the financial impact to an operation. For instance, if botrytis is discovered after harvesting flower, processing through supercritical CO2 extraction will remediate the microbes, making it safe to infuse and for human consumption. However, this does not apply to mycotoxin-producing pests, as those toxins would be concentrated into the cannabis oil. Supercritical parameters of CO2 are defined as temperatures above 31 degrees Celsius (88 degrees Fahrenheit) and pressures above 74 bar (1070 psi). When microbes are present in supercritical fluid environments for prolonged periods of time, sterilization occurs due to cytoplasmic acidification, membrane destabilization and overall physical cell rupture. In layman’s terms, the microbes freak out being present in solution and explode.
This information will enable processors to competently mitigate microbial issues in their operations. Having the capacity and flexibility to navigate through operational issues caused by scientific factors is crucial, and this can be done by utilizing a diverse team of scientists to attack problems.
In conclusion, businesses can successfully interpret data associated with the intricacies of cultivating and manufacturing cannabis products by:
- Understanding technology and methods needed to operate an internal, quality lab to support production.
- Monitoring certificates of analysis (COAs), scrutinizing the individual analyte concentrations rather than just looking at PASS/FAIL.
- Trusting, but verifying the accuracy of third-party testing labs utilizing your own in-house laboratory methods and historical data.
Alisia Ratliff is founder and CEO of the consultancy Victus Capital Ventures.
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