DEVELOPMENT OF ARONIA MITCHURINII AS A SPECIALTY CROP ALTERNATIVE FOR THE DELMARVA REGION.

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: EVANS-ALLEN
• Reporting Frequency: Annual
• Accession No.: 1016197
• Project Start Date: Jun 1, 2018
• Project End Date: May 31, 2022
• Program Code: [(N/A)]- (N/A)

Recipient Organization
University of Maryland Eastern Shore
11868 College Backborne Road
Princess Anne,MD 21853

Performing Department
Natural Sciences

Non Technical Summary
We seek to develop a multidisciplinary area of research in horticultural phytochemistry focused on high-value alternative crops. We will specifically focus on studying the impacts of cultural management (fertility, pest management, etc.) practices along with processing on the phytochemical and nutritional content of the Aronia mitchurinii (also known as back chokeberry) fruit. This plant is a potential next-generation super-fruit containing an antioxidant content up to sixteen times greater than acai berry which is commonly featured in some television programs, and currently widely present in natural and vitamin shops across US. Our unique multifaceted research approach will link agricultural production research with detailed phytochemical analyses and food quality research. Integrating classic agricultural research, phytochemical and food technology training in the laboratory and University outreach settings, this program will facilitate the introduction of new, high-value specialty crops on the Delmarva Peninsula and other US areas. Strongly focused on multidisciplinary research, our goals are to:Develop a horticultural phytochemistry research direction with opportunities centering on an interdisciplinary, collaborative approach for selection and development of high-value specialty crops for small farms and under-served farmer communities.Provide training for students in novel, specialty-crop development including horticulture, phyto- chemistry and food.

Animal Health Component

50%

Research Effort Categories

Basic

30%

Applied

50%

Developmental

20%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
102	1129	2000	50%
102	1129	3100	20%
102	1129	3020	30%

Knowledge Area
102 - Soil, Plant, Water, Nutrient Relationships;

Subject Of Investigation
1129 - Berries and cane fruits, general/other;

Field Of Science
3020 - Education; 2000 - Chemistry; 3100 - Management;

Keywords

phytochemistry

horticulture

cultural management

specialty crops

antioxidants

interdisciplinary approach

training students

black chokeberry

aronia mitchurinii

anthocyanins

polyphenols

flavonoids

small farms agriculture

sustainable farms

Goals / Objectives
Aronia mitchurinii is a specialty horticultural fruit crop that is presently grown in the United States. The parent species which is native to the United States was taken to Eastern Europe, hybridized, and cultivated. The fruit contains the highest known hydrophilic antioxidant content, 15 times greater than the popular tropical fruit, Açai. With an increase in consumer awareness and interest in locally grown foods and especially those with a high nutritional value, markets for specialty crops are expanding. As a result, a fledgling industry based around the high polyphenol-content fruit aronia, is developing in the U.S.. Presently over two million plants are being grown on over 1000 hectares (2470 acres), with the area increasing yearly. Until recently, very little cultural research had been performed on this crop. In 2006 three leading faculty: Primary investigator, Dr. Volkis from UMES and cooperators, Dr. Ristvey (UMCP) and Dr. Brand (UConnecticut) began collaborating to conduct interdisciplinary studies about best cultivation practices, phytochemistry, material science applications and breeding of aronia.Current efforts have been focused on studying the effects of cultural management including fertility, pests and weed management and breeding on phytochemical and nutritional content, as well as processing technology practices to preserve the nutritional and phytochemical characteristics of the fruit, and development of specialty (value added) materials using aronia phytochemicals. To do this we have created an inter-disciplinary approach that addresses the following tasks:Development of criteria for choosing suitable new specialty crops;Correlation between crop growing treatments, breading and nutrition capacity;Development of best practices for production and processing and determination of their influence on nutrition capacity;Development of non-traditional value added applications of the crop, such as material development.The project has four major objectives:Objective 1. Cultural researchCultural studies will take place at Wye REC and cooperating farms, and at UMES. Studies will focus on water use, nutrients, and a comparison of organic vs. traditional production practices on on quality of fruits and antioxidant content.Objective 2. Post harvest quality and fruit phytochemical identification and contribution towards nutritional value, and potential health benefits.These studies will take place at UMES and will involve the development of a comprehensive phytochemical profile fruit from 'Viking' aronia berry. We will determine major and minor polyphenols including anthocyanins, hydroxycinnamic acids, flavonoids, sugars, essential oils, lipophilic antioxidants. Our methods will be as previously described (Xie et al., 2012; Taheri et al., 2013).Objective 3. Aronia Processing3 (a). Experiments will focus on the development of aronia berry anthocyanin extraction techniques using resin extraction: After juicing, aronia pulp can still be utilized for further pigment extraction. Recently published studies have proven the effectiveness of microporous resins in the extraction of anthocyanins from fruit, the first from water extracts of A. melanocarpa pulp (D'Alessandro et al., 2013) and the second from acidified water extracts of grape pumice (Sandhu and Gu, 2013). The goal of this task will be to adapt the technology for aronia and make it suitable for both the juice and pulp extracts. The study will focus on comparative evaluation of different resins and the development of suitable formulations. The effectiveness of four resins: XAD1180N, XAD7HP, XAD761, and FPX66 in the extraction of both anthocyanins and flavonoids from Aronia mitschurinii juice and from pulp extract will be studied using different solvents and conditions. For each formulation we will measure total concentrations of anthocyanins, flavonoids and polyphenols in original juice/extract and in the sorbate. In addition, analysis of purity for all sorbates obtained will be performed using NMR and FTIR methods.3 (b) Experiments will focus on the influence of processing and storage on fruit and products quality. We will determine the concentration of anthocyanins, polyphenols, flavonoids and ORAC factor for aronia juice exposed to temperatures in the range of 60 - 110 °C, and for time periods from 5 to 90 min to determine the phytochemical decomposition thresholds for processing aronia fruit. We will also use thermo-gravimetric analysis to study decomposition mechanisms for aronia juice and extract. Comparison of results as a function of storage time will help to determine the best storage practice for aronia products. For fermentation, we will set up small scale fermentation reactors in the lab utilizing different winemaking technologies. We will take samples from these reactors during the time of vine preparation at all stages of this process and determine the concentration of anthocyanins, polyphenols, flavonoids and ORAC factor. In addition, we will work with wine samples from local winemakers and compare antioxidant quality of wines as a function of technology of its preparation.Objective 4 . Material science researchApplication of aronia extracts in marine antifouling protection. Marine biofilm formation is the accumulation of micro/macro organisms on a submerged structure (Takamatsu, 2003). Solutions such as tributyltin in paint products has proven to be harmful to marine environment and are banned in most of countries. A novel approach for antifouling protection is using antiseptic/antibacterial components and antioxidants to inhibit bacterial growth and stabilize free radicals. Aronia compounds may be effective for antifouling because it contains both antibacterial and antioxidant compounds (polyphenols and flavonoids). Our preliminary tests of aronia juice (Figure 2-2) have shown significant reduction in biofilm precipitation. The goal of this task is to develop effective formulations including polymers and aronia extracts.

Project Methods
Aronia plots will be maintained in collaboration with Wye REC (Dr. Ristvey). New plots will be established for irrigation study. Agricultural disposables will be stocked as needed. In addition, new demonstration plots of aronia with wider than regular distance between plots will be established in UMES' farm. This will allow for demonstration of growing and harvesting of the fruit to farmers and students. Preliminary data has shown that mineral bursts have a pronounced effect on fruit quality, whereas switching from conventional to organic fertilizers had minimal effect if any. Addition of potassium dramatically influenced how fast the bushes grew, but led to reduced concentrations of antioxidants. Nitrogen seemed to have almost identical effects when added in concentrations from 2 to 7 g per year per bush. But addition of 14 g/year/bush led to significant reduction in antioxidant content. Similarly, many other factors, including but not limited to sun/shed regime, irrigation, breading, the soil composition, green house vs. open soil and many others may influence the fruit quality.Aronia will be harvested every August from Wye REC and from collaborating farms including UMES' farm and transported to UMES for study. It will be stored in a deep freezer at -35 deg. C.Phytochemical, processing and material research will be conducted year-round using objectives and methods described above.Sample Preparation: Varying samples of fresh berries will be obtained from experimental plots in Wye Research and Education Center, UMCP and stored in a -35 ` -70 o C freezers. For studies on juice and pulp, aronia berries will be partially defrosted by being placed at room temperature. After they are defrosted they will be ground to a paste using a mortar and transferred to a vacuum filtration unit. Other parts of the berries will be freeze-dried.Pulp Extraction: Typical extractions will be performed on 1 g of solid samples with 25 mL of sequencing grade absolute ethanol and 25 mL of water at 60 o C over a period of 48 hours. Upon completion of the extraction, the extract will be separated from the solvent using vacuum filtration.Anthocyanin measurement: Anthocyanin measurements will be obtained from two dilutions of the same sample. Solutions of 0.025 M potassium chloride and a 0.4 M sodium acetate will be used as the buffers in the dilutions. The 0.025 M potassium chloride solution at a pH of 1 will be used to observe the anthocyanins oxonium and flavilum while the 0.4 M sodium acetate solution at the pH of 4.5 will be used to assess the carbinol form which is colorless. The absorbance of anthocyanins will be determined by measuring each dilution at 520 nm and 700 nm in a Perkin Elmer UV-Vis spectrophotometer.Polyphenols measurement: The measurement of the total phenolic content will be carried out according to Dewanto et la.[i],[ii] Samples will be analyzed spectrophotometrically for the content of phenolic using modified Folin - Ciocalteu colorimetric method.The total flavonoid content will be determined according to Tawekijpokai et al.[iii] Total flavonoid content will be determined using a colorimetric method described above for total phenolics. Briefly, 0.25 mL of the Aronia extract or (+)-catechin standard solution will be mixed with 1.25 mL of distilled water in a test tube followed by addition of 75 µL of a 5% NaNO2 solution. After 6 min, 150 µL of a 10% AlCl36H2O solution will be added and then allowed to stand for another 5 min before 0.5 mL of 1 M NaOH was added. The mixture will be standardized to 2.5 mL with distilled water and mixed well. The absorbance will be measured immediately against the blank at 510 nm.All parameters above will be determined for each sample of juice and pulp separately. In addition, pH, mass ratio of pulp to juice will be determined.Finally we will use wet distillation and hexane extraction to extract essential oils which will be analyzed using GCMS and LCMS technique.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Graduate students, undergraduate students, extension specialists, farmers, scientists Changes/Problems:No changes in research plan and methodology. However, we have already applied for one year of project extension, because we lost lot of laboratory time due to COVID restrictions and it delayed the graduation of involved graduate student. Over this period of time, we will complete all planned experiments and will publish all planned papers. What opportunities for training and professional development has the project provided?Students involved in the project (both those supported by this grant and those involved with support from different programs) were presenting their research on three regional and one national conferences. The PI has presented an invited talk. How have the results been disseminated to communities of interest?We had three meetings with farmers at the Wye Research and Education Center, at Habanera Farm, and at Mill Runs Farm, where we have presented our findings and its practical applications to local farmers community. What do you plan to do during the next reporting period to accomplish the goals?We plan to finish processing of the 2020 cultural management harvest. We plan to process another round of observation for aronia ripeness based on the 2020 harvest. This time, along with monitoring pH, conductivity, and content of major antioxidants, we plan to also control glucose, fructose and sorbitol concentrations over the period of ripeness. We plan to finish our observations of the influence of the temperature on the antioxidant content of aronia juice to formulate best processing conditions. We plan to complete first round of antifouling tests. We plan to prepare and submit three full papers, present on national and regional conferences. The graduate student involved in the project is planning to graduation between 12/2021 and 05/2022.

Impacts
What was accomplished under these goals? We have completed harvesting aronia for cultural management research. So far, we have compared four years of observations and found trends for the best value of nitrogen, best growing condition. We are now processing the last 2020 year of harvesting - before the preparation of paper summarizing this data. We have monitored another year of ripening of aronia, determined the timing for peak of antioxidants and peak of sugars. We have tried to use conductivity of aronia juice as a base for creating a gauge for farmers, but this parameter did not change proportionally to the level of antioxidants. We then tried commercial glucometer with different pH. We found that: •Glucometer reads Glucosides bonded to anthocyanins of aronia juice at slightly acidic pH •At the same pH glucometer does not read glucose content •At neutral pH buffer glucometer reads glucose, but does not read glucosides bonded to anthocyanins. •Using two standard strips, the device may be calibrated for harvesting. We have completed evaluation of four types of polymeric resins for the isolation of powdered antioxidants from aronia and were able to determine the best resin and conditions for the process. We have determined limits of detections and now are working on a draft of a paper to be submitted in May 2021. We have evaluated the influence of the solvent, concentration of polymer and extract, and parameters of spin-coater for the quality of surface in antifouling samples. Samples are now being evaluated for antifouling properties with samples of water from Assawoman Bay. We have created instructional videos for instrumental techniques that are used with aronia research to train new generations of students that will be involved in the project in the future.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 1) V. Volkis. Emerged Environmental Solutions on the Edge of Phytochemistry and Polymer Science: Natures Gifts for Antifouling Protection, Pest Control and Carbon Dioxide Sequestration. University of Maryland Environmental Research Center at Horn Pointe, Cambridge, MD. Invited talk. May 1st 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 1) Brix vs. Anthocyanins Trends in Aronia mitchurinii Super-Fruit over the Ripening Period as a Key for Best Nutritional Quality. Breann. V. Green1*, A. G. Ristvey2, and V. V. Volkis1 , 1-Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD 21853. 2- WYE Research and Extension Center, University of Maryland College Park 20742. ACS MARM regional meeting. Baltimore, MD. June 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 2) Determination of the Composition of Essential Oils and Antioxidants in Holy basil, Argan Oil and Aronia, and their Application in Pest Control. Mark Joseph, Simon Zebelo and Victoria V. Volkis. 1-Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD 21853. ACS MARM regional meeting. Baltimore, MD. June 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 3) PROBING ESSENTIAL OIL EXTRACTS FROM ARONIA MITSCHURINII AND HOLY BASIL AS POTENTIAL ANTIFOULING AGENTS. Carson Cohen1, Barry Teemer1, Evgueni Nesterov2*, Paulinus Chigbu1, Baruch Volkis1, Victoria Volkis1*; 1-Department of Agriculture and Natural Sciences, University of Maryland Eastern Shore, 11868 College Backbone Rd, Princess Anne, MD 21853; 2Department of Chemistry & Biochemistry, Northern Illinois University, 1425 Lincoln Hwy, DeKalb, IL 60115. 2019 UMBC Undergraduate Research Symposium. Baltimore, October 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 4) MONITORING ANTIOXIDANT CONTENT OF ARONIA MITCHURINII JUICE DURING FERMENTATION PROCESS. Awaab Elkarim, Victoria V. Volkis*. 1-Department of Agriculture and Natural Sciences, University of Maryland Eastern Shore, 11868 College Backbone Rd, Princess Anne, MD 21853; 2019 UMBC Undergraduate Research Symposium. Baltimore, October 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: 7) Probing Essential Oil Extracts from Aronia mitschurinii, hemp, and Holy Basil as Potential Antifouling Agents. Teemer Barry, Carson Cohen, Victoria V. Volkis*, Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, Maryland, 2020 Conference of the Northeast Regional Honors Council (NRHC)". Albany, NY

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Students Faculty Farmers Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One graduate student is supported by the project. The training included GLP, report writing, presentations on regional and national conferences, team work, presentations for farmer community and work in phytochemical lab and in two extensions with experimental plots. How have the results been disseminated to communities of interest?Results were presented on regional and national conferences: UMBC regional research symposium, UMES graduate regional research symposium, ARD National biannual meeting, and ACS MERM regional meeting. This project has participated in UMES farm agricultural day, Wye Research and Education Center aronia twilight tour, and during the Small Farms Conference at UMES. This is where the project was dessiminated to communities of rural small farms. In addition, researchers in this project are constantly collaborating with Habanera Farm LLC and MAAGA - Mid-Atlantic Aronia growers Association and often visit these farms to collect harvest samples. What do you plan to do during the next reporting period to accomplish the goals?1) We will conduct the last round of observations for aronia cultural management with temperature days data 2) We will conduct another round of observation for ripening process 3) We will conduct another at least two rounds of fermentation experiments 4) We will prepare and suubmit three papers: one for cultural management, one for resin extraction, and one for the influence of temperature on the antioxidant content of aronia juice. 56) We will prepare a final report for our third and terminal year of this project

Impacts
What was accomplished under these goals? 1) The aronia harvest of 2018 was processed and the following parameters were analyzed: juice from berries, brix, pH, anthocyanins, flavonoids and polyphenol content in juice, as a function of the following factors: - Nitrogen rate measured as g of N/ per year/ per bush; - organic vs. traditional growing, aplication of mineral bursts. The results were also correlated to degrees days. The level of antioxidants as a function of factors tested followed similar trends as in previous years. The concentration of anthocyanins however varied between years and this could be correlated to temperature during the growing season. Cultural management such as nutrient application, mineral amendments, age of plants during the growing season may influence the quality and quantity of phytochemicals in the fruit. Thus, for 2018, as in previous years, nitrogen rate of 3 g/bush/year is an optimal rate in order to achieve the highest antioxidant content. No signifficant difference between phytochemical quality of fruits grown organic vs. regular protocol, while mineral burst helped to slightly increase the antioxidant content. Our 2018 data was well correlated with previously collected. 2) Samples from the 2019 harvest were collected, processed and stored for analysis. 3) We have finished obtaining the data for the optimal thermal processing of aronia harvest and for the l resin extrction process of antioxidants from aronia juice. Data from this research are currently being analyzed for publication. 4) We have so far completed two rounds of experiments with fermentation and wine making with aronia juice with the aim to determine the best procedure that will help to preserve maximum of antioxidants in wine. First two rounds of this experiment were needed to optimize the experimental design and probing scheme. We will run more rounds of fermentation to collect statistically sound data. 5) We have performed first two rounds of observation in the study of aronia ripening process with particular interest on trends for brix and anthocyanins as a function of time. We have observed that the peak of anthocyanin concentration, which is valuable for application in medicine and as a food additive, happens much earlier than the peak of brix, that is valuable for food related applications. During the second round, we added measurements of conductivity with the aim to later construct simple device for farmers that would allow to determine optimal harvesting time as function of application. This device would work for any berries or fruits containingc anthocyanins. We plan to run some more observations, before we publish the data and hopefully patent the idea of the device.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 2) Interdisciplinary Approach for New Specialty Value-Added Crops to Improve Sustainability of Small Farms. V. V. Volkis*, Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD 21853; A. G. Ristvey, WYE Research and Extension Center, University of Maryland College Park 20742. 19th ARD Symposium. Jacksonville, FL. March, 2019
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 3) Correlation between Brix, Antioxidant Content and Total Daylight Hours as a Tool for Determination of Ripeness in Aronia Mitschurinii. Breann Hrechka1, Victoria Volkis1, Andrew Ristvey2. 1Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD 21835. 2University of Maryland Extension, Wye Research & Education Center, P.O. Box 169, Queenstown, MD 21658-0169 . 19th ARD Symposium. Jacksonville, FL. March, 2019
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 5) Natural Extracts from Medical Herbs and Specialty Crops as Effective Pest Management Solution in Organic Agriculture. Mark Joseph, Simon Zebelo, Victoria V. Volkis. 1Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD, 21853. 2018 ABRCMS. Indianapolis, IN.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 7) Probing Polymeric Blends with Natural Extracts from Aronia mitschurinii, Algae and Medical Herbs as Effective and Natural Substitutes of Tributyl Tin (TBT) for Antifouling Protection. Darrick Moore*, Hernan Osorio, Baruch Volkis, and Victoria V. Volkis . 1Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD, 21853. 2018 ABRCMS. Indianapolis, IN.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 9) THE INFLUENCE OF FERMENTATION PROCESS ON THE ANTIOXIDANT CONTENT OF ARONIA MITCHURINII JUICE. Awaab Elkarim, Victoria V. Volkis*. Department of Natural Science, University of Maryland Eastern Shore, Princess Anne, MD. 21853. 2018 UMBC Research Symposium. Baltimore, October 2018.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 10) Long term Monitoring the Influences of Cultural Management on Phenolic Compound Yields in Aronia mitschurinii. Breann Hrechka1*, Dr. Andrew Ristvey2, Dr. Victoria Volkis1 Department of Natural Science, University of Maryland Eastern Shore, Princess Anne, MD. 21853. 2018 UMBC Research Symposium. Baltimore, October 2018.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 11) The Influence of Temperature on the Antioxidants Capacity of Juiced Aronia mitchurinii. Taryn Jones, Dr. Victoria V. Volkis*. Department of Natural Science, University of Maryland Eastern Shore, Princess Anne, MD. 21853. 2018 UMBC Research Symposium. Baltimore, October 2018.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 16) Probing Aronia mitschurinii and Holy Basil as Potential Antifouling Agents. Carson Cohen1*, Hernan Osorio1, , Baruch S. Volkis1, Paulinus Chigbu1, Victoria V. Volkis1 . 1*Department of Natural Science, University of Maryland Eastern Shore, Princess Anne, MD 21853. 10th Annual UMES Research Symposium. Princess Anne, MD. April, 2019
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 20) Studying Ripening Process for Aronia mitschurinii  Brix and Anthocyanins Changes. Breann. V. Green1*, A. G. Ristvey2 , and V. V. Volkis1 , 1Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD 21853; 2 WYE Research and Extension Center, University of Maryland College Park 20742 . 10th Annual UMES Research Symposium. Princess Anne, MD. April, 2019
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 1) Brix vs. Anthocyanins Trends in Aronia mitchurinii Super-Fruit over the Ripening Period as a Key for Best Nutritional Quality. Breann. V. Green1*, A. G. Ristvey2, and V. V. Volkis1 , 1Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD 21853. 2 WYE Research and Extension Center, University of Maryland College Park 20742. ACS MARM regional meeting. Baltimore, MD. June 2019.

Progress 06/01/18 to 09/30/18

Outputs
Target Audience:Students - not only students participated in Aronia project including the one funded by this project, but also all other students working in the project as part of CHEM 498 and CHEM 499 courses, honor program and LSAMP program. As well, this project was presented by our students in forums where other students from across the region were participated. Faculty - this project was presented on One national and two regional conferences over the reporting period. The exposure was on faculty and students. Farmers - this project collaborates with UMES farm, Wye Research and Education Center extension, Habanera Farm LLC, MAGA - mid-atlantic aronia growers association, Midwest aronia growers association. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Currently in Dr. Volkis' lab there is one student supported by this project. Total of two graduate students and 12 undergraduate students are working in the lab. Out of them, one graduate and 8 undergraduate students are working on topics related to Aronia project. This project provide following training for them: - Standard protocols, procedures and manuals were developed for all sandard measurements and sample preparation operations in the lab. Generations of student will be able to use these materials - Students had several field days at WyeREC performing harvesting and sample collection and at-field measurements of yield and brix. They also lerned about using statistics in agricultural research from WyeREC specialist Dr. Ristvey. They learned about buffer capacity of different soils and the role of weather station in this research. - Students have been working on the project and learned how to write ongoing report, paper, presentation and even participated in submission of three grant application over summer months. Students have prepared posters to present on regional conferences. Dr. Volkis participated in visiting local farms growing aronia and had communication with local farmers, collecting issues and topics for the continuation of the research. Five presentations have been accepted to be presented by Dr. Volkis and students on UMES Small Farms Conference. As well,. during summer months Dr. Volkis with two students has prrticipated in collaborative activities with North Illinois University. Students spent there ten weeks of internship and Dr. Volkis was visiting for two weeks. Over that period we have extablished collaborations with NIU specialists in mass spectrometry and liquid chromatography, that helped us to set up first methods for newly installed LCMS instrument. We anticipate to continue this collaboration in anticipation of GCMS and HPLC instruments that are planned to be installed in Dr Volkis' lab over 2018-19 academic year. Over all, during the reporting period, Dr. Vollkis has attended two professional training in instrumentation and method dveloping, and one national conference, whereas students were attending one regional conference and are now preparing to present their accepted to presentation aronia projects at two regional and one national conferencs. How have the results been disseminated to communities of interest?1. Presentations on regional and national conferences, includingSmall Farm Conference, UMBC research Symposium, National Honors Research meeting and ABRCMS. 2. We have currently two papers in preparation (this report only covers 5 months period from May 2018 till September 2018). What do you plan to do during the next reporting period to accomplish the goals?1) Analysis of 2018 harvest samples and comparing it with previous data since 2006 2) Completing heating experiments at 90, and 110 ºC 3) preparing two manuscripts and submitting it to publication. 4) Submitting applications for CBG (resubmission) and to at least two other grants 5) Performing NMR characterization of thermal decomposition of aronia juice and modeling the possible chemical mechanism of decomposition. 6) Collecting another round of berry samples, as they are ripening. Measuring brix, pH, antioxidants and conductivity of solution (function of anthocyanin concentration) over the ripening period to determine parameters of peak of antioxidant content and peak of sweeteness. These parameters are important for determining the optimal harvesting time for different applications of aronia. 7) Preparing formulations containing polymers and aronia extracts for antifouling protection and analyzing its effectiveness. 8) Completing current resin extraction series, publishing these results, then starting working on continued flow process for such extraction.

Impacts
What was accomplished under these goals? 1) The aronia harvest of 2017 was processed and following parameters were analyzed: yeild of juice from berries, brix, pH, anthocyanins, flavonoids and polyphenol content in juice, as a function of following factors: - Nitrogen rate measured as g of N/ per year/ per bush; - organic vs. traditional growing - Aplication of mineral bursts The results are prese ted below as a tumble schemes: Aronia content of antioxidants is much hogher than any other fresh fruit; 40x higher than tomatoes and 15x higher than acai berry. Aronia is a prospective specialty crop for local Delmarva farmers. Over twenty farmers in the Delmarva region and over 500 farmers across US grow Aronia. Cultural management such as nutrient application, mineral amendments, age of plants during the growing season may influence the quality and quantity of phytochemicals in the fruit. Thus, for 2017 harvest we have shown that nitrogen rate of 3 g/bush/year is an optimal rate in order to achieve the highest antioxidant content; there is no signifficant difference between phytochemical quality of fruits grown organic vs. regular protocol; mineral burst helps to slightly increase the antioxidant content of aronia. Our 2017 data is in good correlation with previousely obtained data over the period 2010-2016 2) Samples from the 2018 harvest were collected, processed and propoerly stored 3) As a part of our study of best conditions for aronia pasterization and processing, we have studied decomposition of antioxidants in aronia juice at 80, 100 and 120 ºC as a function of time between 5 min. and 2 hours.We have found that at 80 ºC only about 15% of antioxidants decomposes over a period of two hours. However at 120 ºC already after 5 min. of heating more than 70% of antioxidants are decomposed. Collecting of more data at 90 and 110 ºC is required and planned for the next year. As well, we have found that while the concentration of anthocyanins decreases signifficantly over the heating period, the concentration of simple phenolics increases. This promots us to conclude that most valuable antioxidants, anthocyanins, are not decomposed completely, but converted chemically into simple phenolics. We will conduct HPLC and NMR investigation to confirm this hypothesis and provide chemical mechanism of decomposition. This data would be useful for all crops containing anthocyanins. 4) We have determined limits of detections (LOD) for all UV/Vis measurements techniques we use. This is required to assure that the data lmeasured in the project is within the limit of accurate measurements for the instrument and method used. As well, many journals require this information in order to accept the manuscripts for publication.We use absorbance in UV/Vus spectrometer as a measure and used the standards at different dilutions for the determination. For anthocyanins adenasine 3-Glu was used as a standard. For total polyphenols galic acid was used; and for flavonoids questicin was a standard. 5) We have examined four polymeric resins for its ability to absorb and later release antioxidants from aronia juice. We plan to perform similar experiments with extract from aronia pulp too. This technology will be usefu for production of medica suppliments from aronia.Resins XAD1180N, XAD761, XAD7HP, and FPX66 were obtained from Daw Chemicals. Duplicate measurements of anthocyanins from three trials followed by ANOVA statistical analysis (α=0.01) confirmed results from previous literature that Amberlite FPX66 is the most effective resin of those tested for extraction anthocyanins from fruit material, including Aronia mitchurinnii. Flavonoids and polyphenols were also found in final solutions and showed similar affinities to the resins as anthocyanins. FPX66 is currently the resin of choice for anthocyanin extractions. Efficiency appears to increase as anthocyanin concentration of the original solution is decreased.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Determination of the Composition of Essential Oils and Antioxidants in Medicinal Herbs and Horticultural Crops, and their Application in Pest Control. Mark Joseph1*, Simon Zebelo1 , Victoria V. Volkis1 Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD, 21853. Emory Research Symposium. Atlanta, GA 2018
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2018 Citation: THE INFLUENCE OF FERMENTATION PROCESS ON THE ANTIOXIDANT CONTENT OF ARONIA MITCHURINII JUICE Awaab Elkarim, Victoria V. Volkis*. Department of Natural Science, University of Maryland Eastern Shore, Princess Anne, MD. 21853. 21st UMBC Undergraduate Research Symposium. To be presented October 2018
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2018 Citation: Long term Monitoring the Influences of Cultural Management on Phenolic Compound Yields in Aronia mitschurinii Breann Hrechka1*, Dr. Andrew Ristvey2, Dr. Victoria Volkis1 1Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD, 21853 2 University of Maryland Extension, Wye Research & Education Center, P.O. Box 169, Queenstown, MD 21658-0169. 21st UMBC Undergraduate Research Symposium. To be presented October 2018
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2018 Citation: The Influence of Temperature on the Antioxidants Capacity of Juiced Aronia mitchurinii Taryn Jones, Dr. Victoria V. Volkis* Department of Natural Sciences, University of Maryland Eastern Shore, Princess Anne, MD. 21853 21st UMBC Undergraduate Research Symposium. To be presented October 2018