Source: UNIVERSITY OF ILLINOIS submitted to
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
Funding Source
Reporting Frequency
Accession No.
Grant No.
Project No.
Proposal No.
Multistate No.
Program Code
Project Start Date
Apr 17, 2020
Project End Date
Sep 30, 2021
Grant Year
Project Director
de Godoy, MA.
Recipient Organization
2001 S. Lincoln Ave.
Performing Department
Animal Sciences
Non Technical Summary
Avocado (Persea americana) has become a popular fruit in human nutrition in recent years. According to the USDA, avocado consumption per capita has increased 833% in the last threedecades from 0.8 lb in 1979 to 7.5 lb in 2017, with 2.45 billion pounds of avocados consumed in 2018. Consumers have incorporated avocados into their diets in both raw and processed forms (i.e., raw fruit or cooking oil). Avocado oil also can be found in cosmetics and other personal care products. The rapid increase in the popularity of avocado as a food item has been mainly driven by its association with health and wellness. Avocado pulp is an excellent source of macro and micronutrients, and rich in bioactive lipids. According to the U.S. Nutrition Labeling and Education Act (NLEA) a serving size of avocado (50 g) provides: 80 kcal, 7 g of total lipids (comprised of approximately 67% of mono- and 12% of polyunsaturated fatty acids), 3.4 g of total dietary fiber, and only 0.3 g of sugar. A number of studies have demonstrated that consumption of avocado may improve lipid profile, decrease risk of metabolic syndrome, and aid in weight management, mitigate age-related oxidative DNA damage, and have anti-carcinogenic effects. Although most of the scientific literature has focused on nutraceutical effects of avocado based on its lipid and fatty acid compositionand high antioxidant capacity, recent interest has been devoted to a fatty acid derivative, called persin. The latter is synthesized in idioblast oil cells present in the avocado fruit and leaves with suggested insecticidal and fungicidal activity. Persin is classified as an acetogenin; derived from the biosynthesis of long-chain fatty acids and with similar structure of linoleic acid. In vitro, persin has shown cytotoxic and proapoptotic effects in human breast cancer cell lines. In contrast with the human literature, consumption of avocado by small ruminant (goats and sheep) and monogastric animal species (avians, reptiles, dogs, and cats) is discouraged, due to acute signs of toxicity (i.e., labored breathing, decreased appetite, lethargy, congestion in the lungs, cardiomyopathy, and nephrosis) and eventual death. The toxic effects of avocado have been attributed to the presence of persin, even though this compound has not been quantified in any of those studies. A recent research project in Dr. Godoy's laboratory (PI) in collaboration with Dr. David Sarlah, Assistant Professor of Chemistry have quantified persin in different parts of the raw and cooked avocado fruit (e.g., peel, pulp, and pit). We learned that raw avocado has greater concentrations of persin, and that the peel and pulp are the most concentrated sources of this acetogenin. We are, therefore, interested in evaluating if consumption of avocado oil (extracted from the pulp) could result in unattended health implications due to inherent ingestion of persin. We are proposing to determine whether acute and chronic adult exposure to persin would result in cytotoxic effects in male and female mice, leading to increased oxidative stress and inflammation, and negatively impacting their health and reproductive status. We also plan to evaluate alterations in blood and gastrointestinal metabolic profiling and gut microbiome due to acute and chronic adult exposure to persin using the mouse model.
Animal Health Component
Research Effort Categories

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
Knowledge Area
701 - Nutrient Composition of Food;

Subject Of Investigation
6010 - Individuals;

Field Of Science
1010 - Nutrition and metabolism;
Goals / Objectives
The overall goal of this research is to evaluate the role of persin, a phytochemical in avocado, as a functional or toxic compound using a dose-response approach. The specific aims of this proposal are: 1. To synthesized labeled-persin and determine its metabolism in vivo, 2. To examine the cytostatic and cytotoxic effects of persin in reproductive organs, mammary glands, liver and gastrointestinal tissues, 3. To determine systemic metabolic and gut microbial alterations due to acute and chronic exposure to persin, 4. To evaluate whether metabolic and physiological responses differ when mice are orally dosed with synthetic persin or with equivalent dose of persin using avocado oil. Our central hypothesis is that adult exposure to persin will stimulate oxidative and inflammatory processes in reproductive and gastrointestinal tissues, and will decrease microbial diversity and concentration of short-chain fatty acids, used as proxy of gut health. We also hypothesize that the presence of natural antioxidants, mono- and poly-unsaturated fatty acids present in avocado oil will potential attenuate the cytotoxic effects of persin, or potentially alter its physiological effects in vivo.
Project Methods
Labeled persin synthesis and analysis: High Performance LC/MS will be used to quantify (+)-persin content. Chromatographic separation will be done on a Kinetex® EVO C-18 column (1.7 µm) with a binary gradient from 20% to 90% MeCN in water over twominutes, then 90% to 95% over twominutes at 0.4 mL/minute. Chemical detection will be done with selected ion monitoring (SIM) with ESI ionization on Shimadzu LCMS-2020 instrument. Signal intensities correspond to integrated ion count traces. A five-point linear calibration curve will be built at concentrations of 10, 25, 50, 75 and 100 uM. Experimental design: All experimental protocols will be reviewed and approved by the Institutional Animal Care and Use Committee at the University of Illinois at Urbana-Champaign prior to their initiation. CD-1 female and male mice will be used in a complete randomized design and orally dosed with either vehicle (tocopherol- stripped corn oil, 0% persin), avocado oil or labeled-persin at 10 (low) or 100 (high) mg of persin/kg/day for a total of either 10 days or 30 days to reflect acute vs. chronic exposure to this toxin. All mice will be fed ad libitum a chow diet containing approximately 20% protein, 70% carbohydrate, 10% fat, and 3,85 kcal/g. After 10 or 30 days of receiving the assigned treatments, mice will be euthanized and ovaries, uteri, mammary gland, testes, liver, segments, and content of gastrointestinal tract and blood samples will be collected. All tissues collected will be processed for histological evaluation, whereas ovaries, uteri, mammary gland, and testes will also be evaluated for proliferation and inflammatory markers using immunohistochemistry. Digesta from distal small intestine, cecum, and distal colon will be collected from each mice. DNA extraction, amplification of bacterial, archaeal, and fungal DNA will be performed using targeted primers and a Fluidigm Access Array and sequenced using MiSeq Illumina with v3 reagents. Sequence data will be analyzed using QIIME 2.0. Targeted short-chain and branched-chain fatty acid concentrations will be done using gas chromatography. Serum samples from mice in the same treatment will be pared and analyzed for untargeted serum metabolic profiling at the Metabolomics Center at University of Illinois. Multivariate supervised and unsupervised methods will be used to uncover simultaneous changes of metabolites across experimental factors. Among the supervised strategies that use the experimental conditions as input, k-nearest neighbor, discriminant analysis, and partial least squares methods will be investigated.

Progress 04/17/20 to 09/30/21

Target Audience:Target audiences included the toxicology, food and feed industry, governmental agencies, and research institutes which focus on innovative research approaches to understand the mechanisms of how food components affect human and animal physiology and health. Changes/Problems:COVID-19 was the major hurdle faced. We had to adapt to the situation and think creatively on how to continue to investigate on this topic given all the challenges for the past twoyears. Inability to start new projects, timeline restrictions due to funding for students and general lab costswere also issues that we had to face and adapt to. We believe that we have done the best we could given the circumstances. What opportunities for training and professional development has the project provided?During this project, twograduate students have been trainedin addition to a few undergraduates who have assisted with theproject. How have the results been disseminated to communities of interest?Results of this research have been presented atscientific conferences, and the paper recently published has received a lot of attention due to its novelty and contrubition to the field of nutritional sciences. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

What was accomplished under these goals? Due to COVID-19 we has to change the approach for this research. Starting a new project using animals and large a research team was not an option due to the challenges faced during the pandemic. From this aim we were able to usesynthesized labeled-persin and determine its concentration in various avocade-derived substrates. We also determined its metabolism and nutritional value in vivo using dogs and cats. Lastly, we are evaluating the effects of supplementation of persin on the fecal microbiome of dogs and cats. As these samples were already collected right before the pandemicwe were able to further investigate on this topiceven though we were facing several limitations during that time.


  • Type: Journal Articles Status: Accepted Year Published: 2022 Citation: Dainton, A.N., F. He, T.W. Bingham, D. Sarlah, K.B. Detweiler, H.J. Mangian and M.R.C. de Godoy. 2022. Nutritional and physico-chemical implications of avocado meal as a novel dietary fiber source in an extruded canine diet. J. Anim. Sci. skac026. doi:10.1093/jas/skac026.
  • Type: Journal Articles Status: Other Year Published: 2023 Citation: Dainton, A.N., F. He, K.B. Detweiler, H.J. Mangian and M.R.C. de Godoy. 2023. Avocado meal as a novel and functional dietary fiber source in feline nutrition. In Preparation.
  • Type: Journal Articles Status: Other Year Published: 2023 Citation: Finet, S.E., L. Clark and M.R.C. de Godoy. 2023. Dietary fibers in canine and feline nutrition: A step towards personalized nutrition. In Preparation.

Progress 04/17/20 to 09/30/20

Target Audience:Members of the scientific community in animal sciences and veterinary medicine involved in companion animal and comparative nutrition, and the animal nutrition industry. Changes/Problems:COVID has been the major hurdle of this project, and restrictions on having groups of people working together andacquisiton of laboratory supplies have made it very challenging to start research projects or to successfully continue them. What opportunities for training and professional development has the project provided?The project has allowed for training on analytical techniques involving Goal 1, and hopefully in the near future, as Goals 2-4 will beaccomplished,they will allow for the training and professional development of graduate andundergraduatestudents and a postdoc involved in this project. 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 are hopingto resume the in vivo portion of this project and accomplish Goals 2-4. Our aim would be to have partial reports by late Fall and present an abstract next year in a scientific meeting, followed by a publication in the Fall of 2021.

What was accomplished under these goals? Goal 1 was accomplished and the synthetic labeled-persin was sucessfully produced. Unfortunately because of COVID restrictions in vivo testing has been put on hold.