Progress 09/01/17 to 08/31/18
Target Audience:The audience that we reached during the past twelve months included beginning farmers as part of the non-profit Future Harvest Chesapeake Alliance for Sustainable Agriculture sessions with our commercial partner, Global Aquaponic Systems Co. (GASCO); in addition, educational sessions were conducted with UMES undergraduate/graduate students and faculty and UMD undergraduate students (Resource Sciences & Environmental Technology Depts). Changes/Problems:
What opportunities for training and professional development has the project provided?Over the past year, five undergraduate and graduate students and three research assistants have gone through training and conducted microbiological analyses on water samples and produce in accord with the FDA produce safety rule procedures. They have also advanced their knowledge in food safety extension, soil and food microbiology, and molecular microbiological analyses through on-on-on work with Drs. Hashem, Parveen, and Millner. This mentoring includes class and individual sessions on various topics related to professional skills to agriculture and food science competency topics. They had additional professional development through participation in produce food safety field days that have occurred throughout at UMES and other water quality food safety projects. How have the results been disseminated to communities of interest?Students served as interns in the USDA labs and gained hands-on experience in the sample preparation and operation/interpretation of HPCL analyses of the samples. They also were involved in data analysis, interpretation, and report writing and presentations of experimental results at meetings. Results have been disseminated vis workshops, seminars, meeting, and site visits, and fields days. What do you plan to do during the next reporting period to accomplish the goals?We plan to conduct high tunnel and greenhouse studies on food safety, productivity, and enterprise budget aspects of the aquaponic and hydroponic systems; and publish the research data; also develop a series of fact sheets and videos that show where and how to collect samples from these operating systems for water quality microbial/chemical, and produce safety samples and sanitization of systems, and worker safe handling practices for these production systems. Instructions on cleaning aquaponic units and hydroponic systems in high tunnels and greenhouses Objective 1: Freshly harvested samples of microgreens will be assayed immediately for ascorbic acid, and additional biomass will be frozen immediately in liquid nitrogen and lyophilized for dry weight and other vitamin and carotenoid etc. determinations subsequently. Objective 2: Data Analysis: USDA-ARS, BARC Biometrical Services unit will be consulted to confirm the suitability of design for complex experiments involving factorial, randomized, or block designs with replications, and appropriate internal reference controls. Objective 3: Undergraduate and graduate students will continue to participate in microgreens food safety research and outreach activities. Interns will continue to be mentored and learned a variety of new microbiological techniques and approaches used in evaluating the safety of water, soil amendments, and produce in UMES and USDA-ARS. Objective 4: Food safety plans (FSPs) workshop will be established to produce growers and farmers. .
What was accomplished under these goals?
The intent of the study is to research, educate and provide extension efforts on comparative data in the relationship to phytonutrient density of microgreens and leafy greens and the microbial food safety aspects of their production by water and soil-based systems organically and conventionally. The goal for the extension from UMES was to develop produce food safety education for aquaponics and hydroponics in the U.S. through research on the effects of inoculated research systems in high tunnels. This year the focus of the research project was constructing the fish and shrimp units for research involving inoculation of non-pathogenic surrogates into the systems to determine the length of time they would survive. This also required the development of protocols for sampling and analysis of water, aquatic animals, and horticultural subsections of each research unit. Also, students, research assistance, and scientists tested the procedure for 'curing' the components before animals and plants were introduced into the systems to avoid adverse impacts on animal and plant growth/survival. This concluded in the late fall of 2018 and the High Tunnel housing the aquaponic research units has been readied for early spring 2019 start of replicated inoculation studies (4 inoculated units and 4 control units) within the high tunnel. Soil pots containing the same cultivars of plants transplants into the aquaponics and hydroponics units will also be housed in the high tunnel. Produce from all three production systems (hydroponics, aquaponics, and soil) at microgreens, baby, and mature stages will be harvested and analyzed for phytonutrient composition at UMES. Urban, suburban, school groups, and other researchers and interested farmers will be hosted at on-site field events in collaboration with the UMES and UMD extension teams to share the system technologies, water quality features, and measurement technologies. A microbial research study will be conducted in the summer of 2019 to evaluate the risk of microbial attachment and survival on harvested produce that inadvertently comes into contact with E. coli (non-pathogenic surrogate) contaminated soil, other produce, and water used for post-harvest washing, and work gloves. Sanitation and employee training related to cross-contamination and Food Code will be adapted to hydroponic and aquaponic quick facts sheets, and extension publications. Through this research and extension work, one graduate student and three undergraduate students will be trained on food safety/microbiology in the hydroponics/ aquaponics production environment. Objective 1: Water-based Produce Systems: Microgreens was harvested by cutting with sanitized scissors. Chlorophyll content of harvested microgreens and mature greens was measured either by SPAD meter (Spectrum) or standard extraction/spectrophotometric method. Macro and micronutrients in harvested tissues were measured using standard plant tissue analysis procedures. The phytonutrient content was the focus on total and free ascorbic acid, carotenoids, tocopherols, and phylloquinone. Freshly harvested samples of microgreens were assayed immediately for ascorbic acid, and additional biomass was frozen immediately in liquid nitrogen and lyophilized for dry weight and other vitamin and carotenoid etc. determinations subsequently. Soil-based Produce Systems: Soil samples were collected at experiment initiation and at harvest. Plant tissues were measured for chlorophyll and for the same phytonutrients as for the water-based production systems described above. Sample handling, processing, and analysis, reported with the same involvement of students as described above will be conducted. Objective 2: Water- and soil-based production systems: Produce samples were assayed in a two-stage modification of the shake-rub-shake method to estimate populations of bacteria. Rinsates of LGs was obtained by immersing samples in sterile buffered peptone water in a stomacher bag and manually rubbing them from the outside of the bag 5 min, then soaking 30 min at 4oC. Buffered peptone water was added to stomacher bags containing rinsed samples to achieve a 1:10 dilution, and then homogenized for 2 min. Rinsate and homogenate samples were analyzed separately using standard and enrichment techniques described above. Aliquots (5-ml) of enrichment was transferred to sterile freezer (-80oC) vials for future molecular genetic analysis for target virulence factors, e.g., invA for Salmonella and stx1, stx2 and eae for E. coli, should evidence of natural presence be found. Colonies of Salmonella and E. coli was transferred to suitable culture media, incubated, and stored appropriately for confirmation of identity and genotyping. Individual pathogen isolates recovered was characterized against the background of indigenous isolates by PCR and Rep-PCR and compared to those from water and PL sources to confirm sources. Also, soilless media, water, compost, PL and LG samples were analyzed for pertinent soil and water physico-chemical characteristics using standardized analytical methods. Light irradiance was measured in growth chambers and high tunnels. Objective 3: Summer internships for undergraduate and graduate students to participate in microgreens food safety research. In years 2 and 3 of this project, 8-week summer intern scholarships will be offered competitively not only to UMES students but also all 1890 food and agricultural students (upper level undergraduates with GPA>3.0 and graduates). Evaluation of applicants will be conducted according to fair, open, non-discriminatory rules used by USDA and all participating institutions on the project, and be based on the applicant's response to a request for an essay on a topic of importance in agriculture and food safety, their GPA, and faculty recommendation. These student interns will be mentored and learn a variety of new microbiological techniques and approaches used in evaluating the safety of water, soil amendments, and produce in UMES and USDA-ARS. Objective 4: GAPs/ GHPs training was established to produce growers and farmers' markets. One day Workshop on GAPs/ GHPs practices for micro- and mature leafy greens produce growers and farmers markets at selected 1890s institutions using a curriculum adapted from Cornell University was conducted. The workshops focused on: (1) agricultural practices; (2) water quality (FDA guide); (3) harvesting containers, and sanitizing of harvesting equipment; (4.) shipping logistics which includes refrigeration, (5) health and hygiene/hand washing/toilet, and field sanitation. The GAPs/ GHPs workshops, participants were trained to prepare a food safety plan for their farm/operation. Speakers from state Water Department, FDA inspector, and a local GAPs produce grower, farmers' markets owners (to share successful story on GAPs certification and economics; share in fresh produce safety) contributed to the training modules and presentations at the workshops.
Progress 09/01/16 to 08/31/17
Target Audience:The target audience for this project include aquaculture growers and consumers, fresh produce growers, UMES facility, staff and students, agriculture extension agency, and scientific communities. Changes/Problems:
What opportunities for training and professional development has the project provided?Training and professional development opportunities were provided to 2 facilities member, 5 graduate and 2 undergraduate students to be trained by USDA scientist in growth, harvest and analytical procedures that would be used at the UMES. Two training workshops on aquaponics production were held at a commercial aquaponic facility in Montgomery County, MD, and one at the aquaponics facility of university of District of Columbia in Beltsville, MD How have the results been disseminated to communities of interest?
What do you plan to do during the next reporting period to accomplish the goals?Objective 1: Microgreens will be harvested by cutting with sanitized scissors. Soil samples will be collected at experiment initiation and at harvest. Objective 2: Produce samples will be assayed in a two-stage modification of the shake-rub-shake method to estimate populations of bacteria. Objective 3: Summer internships will be provided to undergraduate and graduate students to participate in microgreens food safety research. Objective 4: GAPs/ GHPs training will be established for produce growers and farmers' markets.
What was accomplished under these goals?
Objective 1: Water-based Produce Systems: Seeds were obtained from the same supplier for all experiments. Sprout seeds were surface sterilized by the FDA recommendations prior to usage. Microgreens were produced in growth chambers in sanitized trays. Water used during the production of hydroponic treatments was tap water and the aquaponics treatments was biofiltered donated by Dr. John Love of the Johns Hopkins University aquaponics project at the Clyburn Arborteum in Baltimore, MD. Water quality measurements (pH, EC, nitrate, ammonia, total carbon, total nitrogen, turbidity, total suspended solids, total aerobic bacteria, free and total chlorine) for both water sources was measured at UMES for each production event and was recorded. In addition, selected plant species for the hydroponic and aquaponics systems in aerated 10-gal containers was setup in a high tunnel at UMES to allow plant growth to proceed to mature full leaf size for comparison with microgreen nutrient content. Light irradiance during the growth period will be measured and recorded in growth chambers and high tunnels. Soil-based Produce Systems: A parallel set of experiments was conducted in large (10L) pots in the same high tunnel as used for the hydroponic/aquaponics mature greens study. Soils were obtained from organically and conventionally managed fields at UMES and amended with poultry litter (PL), on-farm aged PL (semi-composted) and hairy vetch green manure. Samples were analyzed for pH, electrical conductivity, total nitrogen and carbon, water soluble phosphorus, soluble carbon, volatile solids, moisture content, according to standard methods to characterize their stability and maturity. Compost and PL was used to amend experimental soils (11Mg/ha). The same lots of seeds that were used in the water-based systems. Plants were fertilized to reach maturity using conventional or organic fertilizers according to designated treatment using amounts of macro and micro nutrient to equivalency rates. Objective 2: Water- and soil-based production systems: Microgreens and mature greens were produced and water samples were analyzed prior to usage and exited to the production trays. Water and plants was assayed for the presence and enumeration of the following: generic E. coli, EHEC Listeria monocytogenes, Salmonella., Staphylococcus, Pseudomonas, and Enterococcus. Generic E. coli was detected by using standard isolation and identification techniques including pre-enrichment and enrichment protocols adapted to a mini-most probable number (MPN) method for enumeration of samples suspected to have low populations. Polymerase chain reaction (PCR) of enriched samples was used to detect the presence of a target pathogen when culture methods are inconclusive. Appropriate negative controls from both field and laboratory samples was employed to confirm the accuracy of the testing methods. Transfers of microorganisms from the amended soils to foliar LGs was evaluated by inoculating the PL, PL compost, and cut hairy vetch green manure prior to addition to the soil to be placed into the pots. A cocktail of three non-pathogenic E. coli and two attenuated BL-I strains of E. coli O157:H7 resistant to rifampicin and an ATCC non-pathogenic strain of Salmonella was inoculated at 2,000 cfu/g into all organic soil amendments. The inocula simulated modest initial populations of E. coli O157:H7 in amendments was used to determine if the pathogen could survive and transfer to crop by harvest of micro- to baby green and mature stages. Aliquots of inoculated soil amendments was stored at 4oC as the positive controls and all products was assayed at day 0 before and after inoculation and incorporation into high tunnel soil pots (UMES, sandy silt loam) and at each harvest. A total of 960 samples of LGs and 252 soil samples, each assayed for E. coli (generic and attenuated O157:H7) and Salmonella spp. was performed. The total number of samples based on: 6 trts x 5 plant species x 4 replicates x 2 harvests/season x 2 seasons/yr x 2 yrs). LG crops were grown in cool seasons in inoculated, amended soils as required for growth and chard and basil was grown in summer. Cover crop hairy vetch was grown separately and added to the green manure pots. Additional factors was included to determine the effect of low and high levels of pathogen contamination on survival and transfer to the produce. For this 0.1% and 1.0% seeds in separate and independently located microgreen trays was pre-contaminated the E. coli and Salmonella cocktails. Objective 3: Develop and establish an e-learning workshops on microbial food safety, microgreens, leafy greens, aquaponics, and soil amendments for food science, agriculture, microbiology and other undergraduate and graduate students at UMES, and later on will be extended to all other 1890 communities. One hands-on training workshop in Montogemry County and Beltsville, MD each year on: "Fresh Produce Microbiology: Food for Thought Farm-to-Fork" that covers (1) Hazards and Vulnerabilities in Fresh Produce (Microgreens) Production, Handling/Processing, Transportation/Distribution, Retail, and Preparation; (2) Foodborne Hazards: Farm-to-Fork; (3) GAPS/GMPs (4) Legislation/Policy; (5) Communication/Education and; economic analysis. Objective 4: GAPs training and writing of food safety plans (FSPs) were established. Extension specialists at 1890s institutions was trained to teach produce growers and farmers' markets' managers and participants concepts and current best practices to reduce microbial food safety risks in various production systems growing leafy greens. UMES hosted a 1-day "train-the-trainer" GAPs workshop on microgreens and mature leafy greens for Extension Educators across 1890s institutions. Trained Extension Educators assisted and trained farmers and farm market managers and update their food safety training and practices. The workshop included basic farms operations food safety training, writing a food safety plan for a fresh produce farm operation, as well as training tools developed by the Cornell University NIFA-funded project. Extension Educators was provided with USB flash drives that have a food safety plan template. Participants will also be provided with the Cornell University GAPS booklets. Dr. Escobar instructed the GAPs training and the writing of food safety plans.