Source: OREGON STATE UNIVERSITY submitted to
FEEDING SPENT HEMP BIOMASS TO CATTLE: CANNABINOID RESIDUALS, ANIMAL HEALTH, AND PRODUCT QUALITY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
EXTENDED
Funding Source
Reporting Frequency
Annual
Accession No.
1025228
Grant No.
2021-68008-34099
Project No.
ORE01002
Proposal No.
2020-05220
Multistate No.
(N/A)
Program Code
A1701
Project Start Date
Feb 1, 2021
Project End Date
Jun 30, 2024
Grant Year
2021
Project Director
Bionaz, M.
Recipient Organization
OREGON STATE UNIVERSITY
(N/A)
CORVALLIS,OR 97331
Performing Department
Animal & Rangeland Sciences
Non Technical Summary
Our long-term goal is to implement the safe use of hemp byproducts in livestock diets and take full advantage of their nutritional and potential medicinal properties to improve animal health and the quality of animal products. In Oregon, hemp is primarily grown for cannabidiol production. This process yields a high amount of extracted biomass, a byproduct of high nutritive value; however, hemp byproducts are not yet FDA approved to be used in livestock diets. Therefore, we expect to generate data essential for FDA approval and to develop an Extension program on feeding hemp byproducts to livestock.Our primary objective is to generate fundamental data to allow for the legalization of hemp byproducts to be used to feed livestock and to create an Extension program to connect producers with the hemp industry.The Specific Aims are:Determine the residuals of cannabinoids in milk, muscle, and adipose tissue and the effects on health and production in dairy cows fed a ration containing extracted hemp biomass. We will evaluate the effects of replacing alfalfa with extracted hemp biomass in the ration of mid-lactation cows on animal health, milk yield and composition, and levels of cannabinoids in milk, adipose tissue, and muscle.Develop and implement an Extension program to educate livestock producers, general consumers, and policy makers about the potential of feeding hemp biomass byproducts to livestock. With the support of diverse stakeholders, we will conduct surveys to evaluate producers' and consumers' attitudes toward the use of hemp to feed livestock and provide educational programs targeting livestock producers and policy makers about the use of hemp byproducts.
Animal Health Component
100%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3023410101050%
4031730302050%
Goals / Objectives
Determine the residuals of cannabinoids in milk, muscle, and adipose tissue and the effects on health and production in dairy cows fed a ration containing extracted hemp biomass. We will evaluate the effects of replacing alfalfa with extracted hemp biomass in the ration of mid-lactation cows on animal health, milk yield and composition, and levels of cannabinoids in milk, adipose tissue, and muscle.Develop and implement an Extension program to educate livestock producers, general consumers, and policy makers about the potential of feeding hemp biomass byproducts to livestock. With the support of diverse stakeholders, we will conduct surveys to evaluate producers' and consumers' attitudes toward the use of hemp to feed livestock and provide educational programs targeting livestock producers and policy makers about the use of hemp byproducts.
Project Methods
Specific Aim 1: Determine the residuals of cannabinoids in milk, muscle, and adipose tissue and the effects on health and production in dairy cows fed a ration containing extracted hemp biomassSub-aim 1.1. Assessing the effect of feeding extracted hemp biomass on health, performance, and milk quality of dairy cows. This will be achieved by using 20 mid-lactation multiparous Jersey dairy cows from the Oregon State University (OSU) Dairy Center. Ten cows used for the group receiving extracted hemp biomass will be purchased from the Dairy Center, because of the need to euthanize them and landfill the carcass at the end of the study (see below). The cows will be moved into a pen equipped with Calan gates. The animals will receive the typical total mixed ration (TMR) for lactating dairy cows used by the OSU Dairy Center, containing alfalfa hay. Cows will be randomly assigned (based on milk yield, milk composition, and body weight) to two groups. One group will continue to receive the same ration (control), in the other group alfalfa hay will be replaced by extracted hemp biomass.Cows will be fed the diet as TMR with a progressive adaptation to the experimental diet for 7 days prior to starting the experiment. The extracted hemp biomass will be fed for 4 weeks. Cows will be kept in the Calan gate pen for an additional 4 weeks after withdrawal of hemp to assess carry-over effect on feed intake and cannabinoid residuals.Blood will be collected in the morning before feeding from the jugular vein. Blood will be collected just before starting the experiment (day 0) and at day 21 and 42 during the intervention period (i.e., feeding extract hemp biomass). We will also collect two blood samples at 2 and 4 weeks after the intervention period, during the withdrawal period. Serum and plasma samples will be used for the measurement of parameters related to metabolism, inflammation, oxidative stress, and liver function. Blood samples collected at 0, 21 and 42 day of the experiment will be also used to assess white blood cell counting, phagocytosis, and white blood cell differential using specific antibodies against neutrophils and monocytes in association with flow cytometer.Milk samples will be collected during morning and evening milking in 50 mL tubes containing bronopol for analysis of components. At the end of the experiment, cows from the group receiving extracted hemp biomass will be euthanized using barbiturate and the carcass will be landfill.Sub-aim 1.2. Determination of THC and CBD residuals in milk, muscle, and adipose tissue of dairy cows fed extracted hemp biomass. Additional milk samples will be collected in 15 mL tubes and stored at -20°C for analysis of the cannabinoids in the same time points as for the milk component analysis described in Sub-aim 1.1. Sub-cutaneous adipose tissue from the tail-head area and semitendinosus muscle will be obtained via biopsy at the end of the intervention period (day 42), and 2 and 4 weeks during the withdrawal period.Quantification of THC, THCA, CBD, CBDA, and CBN present in milk and adipose and muscle tissues will be performed by a commercial laboratory. The CBD and the THC will be quantified as total CBD and THC calculated as CBD + (0.877 × CBDA) and THC + (0.877 ×THCA) to be compliant with the USDA regulations.Specific aim 2: Develop and implement an Extension program to educate livestock producers, general consumers, and policy makers about feeding hemp biomass byproducts to livestock.?Sub-aim 2.1. Evaluate attitudes of livestock producers regarding the use of hemp byproducts as a feedstuff. To evaluate livestock producers' attitudes toward and potential intent in using hemp byproducts as feed for livestock, a one-time survey will be developed and deployed to livestock producers using the mailing lists of the livestock associations in the state.The survey will explore the attitudes of livestock producers regarding (1) nutritional profile of hemp byproducts (2) price and value of hemp byproducts as a feedstuff, (3) availability of hemp byproducts, (4) ease of access and transport of hemp byproducts, and (5) concerns about regulations. We expect that the results of this survey will highlight when and how livestock producers will consider using hemp byproducts as a feedstuff if approved by the FDA.Sub-aim 2.2. Educate livestock producers about the use of hemp byproducts as a feedstuff. A series of Extension educational meetings will be conducted throughout the state targeting livestock producers. In addition, we expect to provide information at preexisting Extension events (e.g., Beef Industry Tour) and the annual conventions of the Oregon Cattlemen's Association, Oregon Sheep Growers Association, and Oregon Dairy Farmers Association. Events hosted by the GHIC would also be an appropriate venue. The specific locations and content for these educational meetings will be determined in consultation with the stakeholder advisory committee, but briefly we will cover research findings of Specific Aim 1 and other research completed or ongoing at Oregon State University, particularly the types of hemp byproducts, their nutritional profile, cannabinoid content of plants, differences between hemp and marijuana, biological effects of cannabinoids in hemp (CBD and THC), regional availability of hemp byproducts, legal aspects of hemp use in livestock feeding, appropriate practices for including hemp byproducts in the diet including withdrawal periods, and animal health and performance when supplemented with hemp byproducts.?Sub-aim 2.3. Assess the types, volume, and seasonality of hemp byproduct biomass being produced by hemp processors in the state.Hemp byproduct samples will be conducted statewide during the harvest season of 2020 (September/October), focusing on the largest hemp processors. All collected samples will be individually identified, which will allow for matching of the sample to the crop of origin, resulting in a robust dataset with nutritional characteristics and CBD and THC concentrations of the hemp byproducts linked to information about the variety and the location where the plants were harvested.Sub-aim 2.4 Evaluate acceptability to consumers of products from animals that were fed hemp byproducts. As part of the Extension portion of this proposed project, we will include a consumer survey to evaluate the acceptability of consuming products originating from animals that were fed hemp byproducts. This will be a one-time survey deployed to faculty, staff, and students at Oregon State University in an effort to have a diverse demographic pool.This survey will try to understand factors influencing consumer acceptance of products originating from animals that were fed hemp byproducts by focusing on (1) consumers' understating of regulations regarding industrial hemp, (2) perception of risk, (3) environmental and sustainability impact, (4) preference when provide choice, (5) animal welfare, and (6) and producer profitability.Sub-aim 2.5 Educational meeting with policy makers. With the conclusion of the proposed project (research and Extension) our goal is to summarize all data produced by this project and present it to policy makers in local, state, and federal agencies. Like all Extension programming, the information would be science-based, with the goal of providing regulators with sound data that would inform policy creation and rulemaking.

Progress 02/01/21 to 01/31/24

Outputs
Target Audience:The audience for this project are stakeholders, this includes the hemp industry representatives and dairy producers, researchers, and the general public. The means we reached out to the various actors were talks given to conferences and setting where stakeholders invited us to speak. We have also provided up to 3 interviews in various magazines and radio (NPR). We have provided a full report to the advisory committee and to the Oregon Dairy Farm Association. We will present the results of this project to the ADSA annual meeting. Changes/Problems:We put in a Food Use Authorization (FUA) to the FDA to allow to keep the cows after the experiment. After providing to the FDA the data on the cannabinoid residuals in milk, the FDA provided us an FUA and we were able to keep the cows in the herd (originally they had to be euthanized at the end of the study) The issues we had with the experiment were: the cows did not eat all the spent hemp biomass provided. The diets offered to the cows in the hemp group contained approx. 13% spent hemp biomass (as dry matter)but the cows consumed on average 7.45±3.25% (range between cows was 5.0-10.1%) of spent hemp biomass in the diet that corresponded to 1.22±0.53 kg of spent hemp biomass a day (range between cows was 0.8-2.9 kg/d). one animal in the hemp group had to be removed from the final dataset because the cow learned to eat the feed on a control cow's Calan gate; thus, we moved the cows to a sick cow/calving pen where we could feed the cows individually. Because of this, the cows had an increase in somatic cells in milk (i.e.., subclinical mastitis) and became very obese. What opportunities for training and professional development has the project provided?The project provided the opportunity to train: - 1 PhD student - 1 bio technician - 1 intern - 6 undergraduate students All worked on the in vivo part of the experiment (except the PhD, which is working on the analysis of samples and interpretation of the data) How have the results been disseminated to communities of interest?Yes, via several conference presentations and a peer-reviewed publication (seedoi: 10.3168/jds.2023-23829). What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The in vivo study was carried out between May and July 2021 and all samples were collected and the majority of samples were analyzed. Objective 1: we measured cannabinoids in milk collected at the end of the intervention period (4 weeks of feeding spent hemp biomass) and after 4 weeks of spent hemp biomass withdrawal and found cannabinoids in milk during the intervention period but all cannabinoids disappear in the samples of milk collected after 4 weeks of withdrawal. Objective 2: we collected all the data proposed and we are now drafting the manuscript. Completed data are: Nutritional analysis of the spent hemp biomass Measurement of blood parameters related to metabolism, inflammation, oxidative stress, and liver function Milk yield and analysis of milk components Feed intake and amount of hemp eaten by the cows Body condition score and body weight every 2 weeks Complete white blood count, leukocytes migration, and leukocytes phagocytosis via flow cytometer Cow activity and body temperature via Afimilk and smaXtec Determination of the fatty acid composition of the milk

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Bionaz M. 2020. Feeding Spent Hemp Biomass to Dairy Cattle. Invited speaker for the virtual 2021 Tri-State Dairy Nutrition conference, April 19-21, Columbus, OH
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Irawan A, Puerto-Hernandez GM, Ford HR, Busato S, Ates S, Cruickshank J, Ranches J, Estill CT, Trevisi E, Bionaz M. Feeding spent hemp biomass to lactating dairy cows: Effects on performance, milk components and quality, blood parameters, and nitrogen metabolism. J Dairy Sci. 2024 Jan;107(1):258-277. doi: 10.3168/jds.2023-23829. Epub 2023 Sep 9. PMID: 37690708.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Irawan A., Puerto-Hernandez G., Pearce C., Torres M.A., Grismer B., Ates S., Cruickshank J., Ranche J., Bionaz M. Feeding spent hemp biomass to late-lactating dairy cows: effects on performance, milk production, milk quality, and methane emission. 2022 ADSA Annual Meeting
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Irawan A., Ford H. R., Busato S., Ates S., Cruickshank J., Ranche J., Trevisi E., Bionaz M. Feeding spent hemp biomass to late-lactating dairy cows does not affect the immune system but has a minor effect on metabolism and inflammation. 2022 ADSA Annual Meeting
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Irawan A., Nosal D.G., Puerto-Hernandez G., van Breemen R., Ates, S., Bionaz M. Transfer of cannabinoids in milk from spent hemp biomass fed to dairy cows. 2022 ADSA Annual Meeting
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Irawan A., Parker N., Nosal D., van Breemen R., Bionaz M., Ates S. Transfer of cannabinoids compounds from spent hemp biomass fed to lambs in adipose tissue and muscle. 2022 Cannabis Research Conference
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Nosal D., Irawan A., Bionaz M., Ates S., Muchiri R.N., van Breemen R. Analysis of Cannabinoids in Milk from Cows Consuming Spent Hemp using Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry. 2022 Cannabis Research Conference


Progress 02/01/22 to 01/31/23

Outputs
Target Audience:The target audience are scientists, hemp industry, and policy makers Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We trained 1 PhD student, 8 undergraduate students, 1 biotechnician, and 1 research intern through the project How have the results been disseminated to communities of interest?See above about news articles; however, most of the results were presented at conferences such as ADSA Annual Meeting and the Cannabis Research Conference. Results were also provided to agencies that added some funding to the project, such as theOregon Beef Council What do you plan to do during the next reporting period to accomplish the goals?We plan to complete the following analysis and papers: cannabinoids residulas in tissues --> 1 manuscript including the cannabinoids residuals in the various tissues and milk plus the pharmacokinetic of cannabinoids; microbiota in the rumen --> 1 manuscript with the data on the microbiota plus volatile fatty acids in rumen content transcriptome of the liver --> 1 manuscript

Impacts
What was accomplished under these goals? Data from this project have been used to inform the public as the PI and co-PIs have been interviewing by several magazines, here a partial list of the news articles published where the project/study was mentioned: Researchers study spent hemp biomass as animal feed. By GEORGE PLAVEN Capital Press Mar 26, 2021 Updated Mar 26, 2021 Hemp: the green crop tied down by red tape in the US. By Betsy Reed Editor, Guardian US Turning a CBD farming byproduct into feed for cows, chickens and sheep. By Jes Burns. OPB FDA: Safety concerns delaying hemp animal feed approval. By Emma Penrod. Feed Strategy Data from the project were used to submit a USDA NIFA AFRI Foundational proposal The in vivo project was concluded in Summer 2021. The second part of the objective 1 (efefct on health and production) has been completed and all the data are being submitted for publication. The first part of Objective 1 (residuals of cannabinoids) are being performed in all listed tissues. The analysis has been completed for the milk and the data partly publisehd as confernece abstracts. The second objective has been partly accomplished but still need to be completed and no publications exist yet.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Irawan A., Puerto-Hernandez G., Pearce C., Torres M.A., Grismer B., Ates S., Cruickshank J., Ranches J., Bionaz M. Feeding spent hemp biomass to late-lactating dairy cows: effects on performance, milk production, milk quality, and methane emission. 2022 ADSA Annual Meeting
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: . Irawan A., Ford H. R., Busato S., Ates S., Cruickshank J., Ranches J., Trevisi E., Bionaz M. Feeding spent hemp biomass to late-lactating dairy cows does not affect the immune system but has a minor effect on metabolism and inflammation. 2022 ADSA Annual Meeting
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Nosal D., Bionaz M. Muchiri R. Van Breemen R.B. 2021. Analysis of Cannabinoids in Milk from Cows Consuming Spent Hemp using Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry. ASMS Conference on Mass Spectrometry and Allied Topics.
  • Type: Journal Articles Status: Submitted Year Published: 2023 Citation: A. Irawan1, G.M. Puerto-Hernandez, H.R. Ford, S. Busato, S. Ates, J. Cruickshank, J. Ranches, C.T. Estill, E. Trevisi, and M. Bionaz. Feeding spent hemp biomass to lactating dairy cows: effects on performance, nitrogen metabolism, milk components and quality, and blood parameters. Journal of Dairy Science (submitted)


Progress 02/01/21 to 01/31/22

Outputs
Target Audience:The audience for this project are stakeholders. This includes hemp industry representatives and dairy producers, researchers, and the general public. We reached out to the various actors through conference talks, and other settings where stakeholders invited us to speak. We have also provided up to 3 interviews in various magazines and radio (NPR). We have provided a full report to the advisory committee and to the Oregon Dairy Farm Association. We will present the results of this project to the ADSA annual meeting. Changes/Problems:We put in a Food Use Authorization (FUA) to the FDA to allow to keep the cows after the experiment. After providing to the FDA the data on the cannabinoid residuals in milk, the FDA provided us an FUA and we were able to keep the cows in the herd (originally they had to be euthanized at the end of the study) The issues we had with the experiment were: • the cows did not eat all the spent hemp biomass provided. The diets offered to the cows in the hemp group contained approx. 13% spent hemp biomass (as dry matter) but the cows consumed on average 7.45±3.25% (range between cows was 5.0-10.1%) of spent hemp biomass in the diet that corresponded to 1.22±0.53 kg of spent hemp biomass a day (range between cows was 0.8-2.9 kg/d). • one animal in the hemp group had to be removed from the final dataset because the cow learned to eat the feed on a control cow's Calan gate; thus, we moved the cows to a sick cow/calving pen where we could feed the cows individually. Because of this, the cows had an increase in somatic cells in milk (i.e.., subclinical mastitis) and became very obese What opportunities for training and professional development has the project provided?The project provided the opportunity to train: - 1 PhD student - 1 bio technician - 1 intern - 6 undergraduate students All worked on the in vivo part of the experiment (except the PhD, who is working on the analysis of samples and interpretation of the data) How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?We plan to accomplish: - Objective 1: We will measure cannabinoids in milk samples collected during the rest of the experiment (252 samples) to determine the true disappearance of cannabinoids in milk. We will also measure cannabinoid residuals in the adipose, muscle, and liver tissues collected at the end of the intervention period and during the withdrawal (2 times for adipose and muscle tissue and once for liver tissue). We will be also measured cannabinoids in blood collected every three hours for 24h during the intervention period. We plan to draft the manuscript before the end of 2022. - Objective 2: we have completed all the analysis and we will submit the manuscript with those data by Summer 2022.

Impacts
What was accomplished under these goals? The in vivo study was carried out between May and July 2021 and all samples were collected and the majority of samples were analyzed. Objective 1: we measured cannabinoids in milk collected at the end of the intervention period (4 weeks of feeding spent hemp biomass) and after 4 weeks of spent hemp biomass withdrawal and found cannabinoids in milk during the intervention period but all cannabinoids disappear in the samples of milk collected after 4 weeks of withdrawal. Objective 2: we collected all the data proposed and we are now drafting the manuscript. Completed data are: 1. Nutritional analysis of the spent hemp biomass 2. Measurement of blood parameters related to metabolism, inflammation, oxidative stress, and liver function 3. Milk yield and analysis of milk components 4. Feed intake and amount of hemp eaten by the cows 5. Body condition score and body weight every 2 weeks 6. Complete white blood count, leukocytes migration, and leukocytes phagocytosis via flow cytometer 7. Cow activity and body temperature via Afimilk and smaXtec 8. Determination of the fatty acid composition of the milk

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Bionaz M. 2020. Feeding Spent Hemp Biomass to Dairy Cattle. Invited speaker for the virtual 2021 Tri-State Dairy Nutrition conference, April 19-21, Columbus, OH