Item No. 1 of 1

INVESTIGATOR: Ray, A. J.; Antonious, G.; Dasgupta, S.; Durborow, R.; Cochran, N.; Weibel, C.; Silitonga, M.; Malone, R.

PERFORMING INSTITUTION:
KENTUCKY STATE UNIVERSITY
FRANKFORT, KENTUCKY 40601

INTEGRATED APPROACH FOR SUSTAINABLE AQUACULTURE AND AGRICULTURE PRODUCTION WITH THE USE OF RECLAIMED WATER FROM RURAL COMMUNITIES

NON-TECHNICAL SUMMARY: It is estimated that there are over 16,000 publicly owned wastewater treatment plants in the U.S. that treat 35 billion gallons per day, and produce 6.5 million metric tons of dried sewage sludge. Only 6% of the municipal wastewater is reused. This mean that millions of gallons of reclaimed water are released daily back to the river systems without any further use. This project support the development of a cutting edge technology in which wastewater resources and biosolids are used to produce commercial levels of fish and plants. The use of reclaimed water for aquaculture is an innovative approach to secure food production through the integration of decommissioned wastewater treatment plants with sustainable agricultural technologies. With better methods for processing wastewater, many municipalities are now building new, larger facilities, and decommissioning the old ones, many of which have tanks and ponds that could be converted for fish culture. Many are being needlessly demolished when recycling them as fish production facilities could save the community demolition costs, create new jobs and generate revenue. The new facilities are being built adjacent to the old, and would allow reclaimed water to be used for aquaculture. One concern of using reclaimed water for food production is the chemicals that could be present in the water especially persistent organic pollutants such as endocrine-disrupting compounds (EDCs). There are different methods for the treatment of reclaimed water. During the final treatment stage, water could be disinfected by ozone or ultraviolet light (UV). The goal of this integrated project is to use reclaimed water for aquaculture and agriculture enterprises for rural municipalities and their limited resource farmers. Specific objectives are to: 1) evaluate and compare fish production, water quality and bioeconomics using ozone-treated, UV-treated, and UV combined with ozone-treated reclaimed water technology; 2) biomonitor fish for contaminants using reclaimed water; 3) biomonitor plants for contaminants using biosolids; 4) develop a demonstration farm and provide workshops for dissemination and transfer of sustainable technology to rural communities, and 5) develop on-line course on theory and application of reclaimed water treatment. In this project we want to prove that fish culture in reclaimed water and plants grown in biosolids are safe for human consumption. This project will compare the removal and bioaccumulation of EDCs from reclaimed water treated. The results of this project will provide technological information for using reclaimed water facilities for agriculture enterprises with UV and ozone. In addition, the results will be an important and critical step in understanding the accumulation of EDCs within the particular food when reclaimed water or biosolids are used. This project will serve as a model to other universities, federal and state agencies and stakeholders involved in water reuse and agriculture by providing bench-mark information and educational opportunities pertinent to the advancement of sustainable agriculture production systems that uses reclaimed water and biosolids.

OBJECTIVES: The goal of this project is to use reclaimed water for aquaculture and biosolids for agriculture enterprises for rural municipalities and their limited resource farmers. Specific objectives are: 1) To evaluate and compare fish production, water quality and bioeconomics using ozone-treated, UV-treated, and UV combined with ozone-treated reclaimed water technology; 2) To biomonitor fish for contaminants using reclaimed water; 3) To biomonitor plants for contaminants using biosolids; 4)To develop a demonstration farm and provide workshops for dissemination and transfer of sustainable technology to rural communities, and 5) To develop on-line course on theory and application of reclaimed water treatment. The results of this project will provide technological information for using reclaimed water facilities (RWF) for agriculture enterprises. As by-products from conventional RWF, reclaimed water will be used for aquaculture in combination with sewage sludge that will be introduced as an alternative agricultural resource. The project will provide information to verify the safety of the crops grown from reclaimed water and biosolids by evaluating contaminants in fish, fruits, soil and water. A key emphasis will be to monitor endocrine disrupting compounds in fish culture in reclaimed water and plants grown in biosolids. The use of reclaimed water and biosolids to augment safe and healthy food supplies will have a significant potential for helping to relief the global food crisis in a changing world. In addition, this novel research information will be delivered by holding workshops in the demonstration farm that promotes rural development. This new way to produce food and income will be outreached to rural communities by helping individuals learn alternative ways of sustainable agriculture. Rural communities could create jobs by embracing the use of reclaimed water and biosolids for agricultural production. Furthermore, the transfer of knowledge generated from this project will be incorporated into an online course. This course will expand and diversify our curriculum at KSU for teaching students. This project will create jobs for a Co-Investigator, a graduate student and two undergraduate workers and expand the KSU curriculum to include theory and application of reclaimed water treatment thereby attracting a broader base of students. These persons will be qualified to pursue further education and careers in agriculture, aquaculture, environmental science, economics and education and to increase the diversity of the U.S. food and agricultural science base. Funding a demonstration farm and learning center will greatly expand the research, teaching and extension capabilities of the aquaculture and environmental programs and other related programs at KSU. Fish and plants from these operations would provide steady supplies of consumer-safe products and alleviate use of groundwater. Reclaimed water and biosolids management alternatives have the potential for expanding the Nation's food supply through reuse of these resources.

APPROACH: Reclaimed Water Experiment: will monitor fish growth and survival and water quality in tanks with flow-through reclaimed water (a.k.a. disinfected secondary effluent) under experimental and commercial conditions. This experiment will use hybrid striped bass and paddlefish as test species. The experiment will be conducted at Frankfort and Winchester Reclaimed Water Facilities. The Frankfort Reclaimed Water Facility (FRWF) uses ozone at the final disinfection treatment stage while the Winchester Reclaimed Water Facility (WRWF) uses UV irradiation. We will install an ozone generator at WRWF for advanced oxidation processes (UV/Ozone). Fish will be culture at WRWF in decommissioned tanks filled with UV-treated and UV/Ozone treated. In FRWF, fish will be culture in ozone-treated reclaimed water. Data on fish survival, growth and water quality will be analyzed using a mixed ANOVA procedure. Economic and financial analyses will be generated. Indicators such as production efficiency (i.e. gross yield), input use efficiency (i.e. feed conversion ratio), and profitability will be used to measure performance of aquaculture operations. Fish Biomonitoring for Contaminants Experiment: will biomonitor fish grown at FRWF and WRWF for contaminants. For the long-term experiment, fishes from the UV-treated and UV/Ozone-treated group from WRWF, and from the ozone-treated from FRWF will be stocked in cages and pond and biomonitor for one year. Fishes will be analyzed for bioaccumulation of endocrine disrupting compounds and heavy metals. Other sources of contamination that will be analyzed are: lived food, prepared diet, and water sample. Data will be analyzed using a mixed ANOVA procedure. It is anticipated that contaminants if present might be higher in fish from the UV-treated group than those in other disinfectant treatments. Plants Biomonitoring for Contaminants Experiment: will biomonitor plant grown at WRWF for contaminants. The experiment design is a 4 x 3 randomized complete block design with four treatments including the two high-value niche crops (poblano pepper and Roma-type tomato) and two soil types (biosolid Class A from WRWF and no sludge soil). Other sources of contamination that will be analyzed are: soil, seedling plant, and water sample. Demonstration Farm for Extension Services: will provide formal training, demonstrations and workshops for dissemination and transfer of sustainable technology to rural communities. The aim of this model farm is to increase food production by using existing resources such as decommissioned reclaimed water facilities. This innovative approach will promote sustainable agriculture systems by producing safe and healthy food sources and by increasing consumer awareness of the potential of reclaimed water and biosolids for agriculture purposes. On-line Course Development: will develop a course on theory and application of reclaimed water treatment and include the most recent water reuse sustainability practices and other sustainable options for management of the solids at RWF. Information generated by the research part of this integrated project will be incorporated into the course.

PROGRESS: 2012/09 TO 2016/08
Target Audience:Potential fish producers, current fish producers, municipalities, other scientists. Changes/Problems:The laboratory in Arizona (The Arizona Lab for Emerging Contaminants) we have been using was unable to analyze the endocrine-disrupting compounds in our samples. The samples were sent tothis laband they never produced any results. They were asked about the samples repeatedly for about one year, and probably lost them or destroyed them somehow, as they did not send any data. What opportunities for training and professional development has the project provided?A Master of Science student was able to work towards a thesis research project. Her thesis is still pending, but she collected much of the data for this project. One undergraduate student was involved in the project during this reporting period as a student research assistant. How have the results been disseminated to communities of interest?A farmer has continued to use the reclaimed water facility to grow fish, helping to demonstrate the use of the techniques developed by this project. A conference paper and a posterwere given at the Aquaculture 2016 Conference, both regarding this project. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

IMPACT: 2012/09 TO 2016/08
What was accomplished under these goals? To meet the project objectives, hybrid striped bass were harvested from the lagoon cages and processed for analysis. Samples of fish were filleted and fillets were sent to the University of Arizona's Laboratory for Emerging Contaminants to be analyzed for metals and endocrine disrupting compounds. We found that there were no signiicant differences between treatments regarding the concentration of the metals of concern in this project. None of the metals concentrations in fishcame close to FDA action limits, although zinc was high in the prepared diets used.

PUBLICATIONS (not previously reported): 2012/09 TO 2016/08
Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Cuevas-Uribe R., Mims S.D. 2016. Bioaccumulation of contaminants in market-size hybrid striped bass grown in reclaimed water. Abstract Book of the Aquaculture 2016 Conference in Las Vegas, NV, USA.

PROGRESS: 2014/09/01 TO 2015/08/31
Target Audience:Potential fish producers, current fish producers, municipalities, other scientists. Changes/Problems:The project was transferred to Andrew Ray as the new PD. Andrew started his employment at KSU during this reporting period (September 16, 2014). The person who holds the lease at the Winchester WWTP has expressed hostility towards the project, indicating that he would like to keep the equipment at the facility which was purchased under this project. It has therefore been decided that all equipment purchased through this project will be removed. The likelihood of conducting a workshop on the property is low. KSU has implemented a policy in which new academic courses are strongly discouraged due to financial considerations. The likelihood of being able to develop the online course originally proposed is low. What opportunities for training and professional development has the project provided?A graduate student was hired in August 2015. This student is now responsible for compiling, collecting, and analyzing the data regarding the hybrid striped bass portion of the project. One undergraduate student was involved in the project during this reporting period. How have the results been disseminated to communities of interest?One farmer is involved in the project. This person holds a lease on the treatment facility site and has been involved in the culture of fishes at that site. A professional conference abstract was submitted and accepted by the graduate student on the project. Four articles have been published in the KSU Aquaculture Division's weekly newsletter Aquativities and these stories have been posted on the KSU Aquaculture Facebook page with which farmers interact. What do you plan to do during the next reporting period to accomplish the goals?We plan to harvest hybrid striped bass from the cages in the lagoon and finalize the contaminants analyses. We then plan to start preparing a manuscript to describe the results of the project.

IMPACT: 2014/09/01 TO 2015/08/31
What was accomplished under these goals? To address project objectives, paddlefish were harvested from the cages in lagoon two at the Winchester Wastewater Treatment Plant (WWTP) in November 2014. After the winter, hybrid striped bass were then moved from the concrete tanks into the lagoon cages in May 2015, where they will be monitored until September 2015. The harvested paddlefish and a sample of hybrid striped bass, as well as previously collected fish, feeds, and water from the project were processed and sent to the University of Arizona's Laboratory for Emerging Contaminants where they are being analyzed for metals and endocrine disrupting compounds concentrations. Metals data was finalized for these samples, but endocrine disrupter data are still pending.

PUBLICATIONS: 2014/09/01 TO 2015/08/31
Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Gamez E.R., Cuevas-Uribe R., Mims S.D., Ray A.J. Tissue concentrations of heavy metals and endocrine disrupting compounds in hybrid striped bass grown in a municipal reclaimed water aquaculture facility. Aquaculture 2016, Las Vegas, Nevada, USA.

PROGRESS: 2013/09/01 TO 2014/08/31
Target Audience: Potential fish producers, municipal utilities, other scientists. Changes/Problems: Dr.Mims, the PD on the project has retired. Dr. Cuevas, the primary Co-PD has taken other employment. What opportunities for training and professional development has the project provided? Four undergraduate students worked and learned about aquaculture production and the potential of using wastewater treatment plants in production practices. How have the results been disseminated to communities of interest? Presentations were made at national and state meetings to producers and other scientists. Two journal articles have been submitted and accepted. What do you plan to do during the next reporting period to accomplish the goals? The paddlefish in the lagoon cages will be harvested and tissue analyzed for changes in contamination concentrations. Hybrid striped bass will be harvested from the primary exposure tanks. Fish will be removed and submitted for contamination analysis. A portion of the remaining hybrid striped bass will be transferred to the cages for monitoring.

IMPACT: 2013/09/01 TO 2014/08/31
What was accomplished under these goals? A floating cage system was constructed in Lagoon Two at the Winchester Wastewater Treatment Plant(WWTP). Paddlefish from the Year 1 study on bioaccumulation in tanks were transferred into the cages where they will be monitored until October 2014.After the paddlefish were transferred, the tanks were cleaned and stocked with Hybrid Striped Bass in early July to begin phase two of the contaminant bioaccumulation study. Hybrid Striped Bass were also stocked into a 140,000 gallon tank at the Midway WWTP as well as a 30,000 gallon tank at the Frankfort WWTP. Feeding and water quality analyses are ongoing. During the reporting period biosolids were transported from the Winchester WWTP to Happy Jacks Produce Farm, where the vegetables are being grown for contamination analysis. Water samples from the irrigation source (Elkhorn Creek) are being taken weekly and combined to form a composite sample.

PUBLICATIONS: 2013/09/01 TO 2014/08/31
1. Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Cuevas-Uribe, R., & Mims, S. D. (2014). Investigation in reuse of decommissioned wastewater facility and reclaimed water for culturing paddlefish fingerlings. World Aquacult. Soc. Accepted jwas.13367.
2. Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Cuevas-Uribe, R., & Mims, S. D. (2014). Contaminant bioaccumulation in paddlefish and hybrid striped bass juveniles cultured in reclaimed effluent water at a decommissioned wastewater treatment facility. Journal of Applied Ichthyology Accepted.
3. Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Auberry, P., Cuevas-Uribe, R., & Mims, S.D. (2014, February). Comparative monoculture of hybrid catfish and paddlefish in Kentucky. Presentation at Aquaculture America, Seattle, Washington.
4. Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Juanich, G., Cuevas-Uribe, R., & Mims, S.D. (2014, February). Water reclamation for aquaculture through ozone and/or ultraviolet radiation treatment. Presentation at Aquaculture America, Seattle, Washington.
5. Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Squadrito, A., Cuevas-Uribe, R., & Mims, S.D. (2014, February). Effects of live versus commercially available prepared diets on the growth and survival rate of post larval paddlefish, Polyodon spathula. Presentation at Aquaculture America, Seattle, Washington.
6. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Cuevas, R. and Mims, S. (2013, September). Reuse technology opportunities at decommissioned wastewater treatment facilities using reclaimed water. 28th Annual WateReuse Symposium. Denver, CO.
7. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Mims, S. D. and Cuevas-Uribe, R. (2013, September). Contaminant bioaccumulation in paddlefish and hybrid striped bass fingerlings cultured in reclaimed effluent water at a decommissioned Wastewater Treatment Facility. Diversification in Inland Finfish Aquaculture II, Vod?any, Czech Republic.
8. Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Henderson, P.W., Cuevas-Uribe, R., & Mims, S.D. (2014, February). Preliminary evidence on the control of off-flavor and cyanobacteria in hybrid catfish ponds through biomanipulation using paddlefish. Presentation at Aquaculture America, Seattle, Washington.

PROGRESS: 2012/09/01 TO 2013/08/31
Target Audience: Target audience is municipalities who have decommissioned facilities and arewilling to invest funds to recondition their plant for aquaculture and agriculture production. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The project has provided training to two graduate students and three undergraduate students in the reuse of a decommissioned facility for aquaculture. How have the results been disseminated to communities of interest? Results are being disseminated via our website www.paddlefishfarming.com as well as through abstracts and presentations at scientific conferences. What do you plan to do during the next reporting period to accomplish the goals? We plan to use phase-II paddlefish in a production study using reconditioned lagoon ponds and monitor any bioaccumulation of contaminants in the fish tissue at harvest. Tomatoes and peppers will be planted in grade a biosolids to monitor growth and bioaccumulation of contaminants. Bass production and water quality will be monitored using ozone-treated, UV-treated, and UV combined with ozone-treated reclaimed water technology.

IMPACT: 2012/09/01 TO 2013/08/31
What was accomplished under these goals? Research was completed on phase-II production of paddlefish production and water quality using ozone-treated, UV-treated, and UV combined with ozone-treated reclaimed water technology. Fish tissue samples have been analyzed and indicate safe levels under FDA guidelines for contaminants such as heavy metals and organochlorine products. The decommissioned facilities has been re-purposed as a demonstration farm and provided two workshops for professionals and hosted one field trips for high school students.

PUBLICATIONS: 2012/09/01 TO 2013/08/31
1. Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Cuevas-Uribe, R. & Mims, S. D. 2013. Investigation in reuse of decomissioned wastewater facility and reclaimed water for culturing paddlefish fingerlings. Journal of the World Aquaculture Society.
2. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Cuevas-Uribe, R. & Mims, S. D. 2013. Investigation in reuse of decomissioned wastewater facility and reclaimed water for culturing paddlefish fingerlings. Journal of the World Aquaculture Society, Accepted JWAS-13-367. Presentations Cuevas-Uribe, R. & Mims, S. D. 2013a. Reclaimed water aquaculture: an innovative sustainable technology of producing fish. Pages 116. 17th Biennial Research Symposium. Association of 1890 Research Directors, Jacksonville, Florida.
3. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Cuevas-Uribe, R. & Mims, S. D. 2013. Reuse technology opportunities for aquaculture at decommissioned wastewater treatment facilities for paddlefish fingerling culture. Pages 229. Aquaculture 2013. World Aquaculture Society, Nashville, Tennessee.
4. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Cuevas-Uribe, R. & Mims, S. D. 2013. Reuse technology opportunities for aquaculture at decommissioned wastewater treatment facilities using reclaimed water. 28th Annual WateReuse Symposium. WateReuse Association, Denver, Colorado.
5. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Henderson, P., Cuevas-Uribe, R. & Mims, S. D. 2013. Can paddlefish control the off-flavor causing cyanobacteria in hybrid catfish ponds? KAS 99th Annual Meeting. Kentucky Academy of Science, Morehead, Kentucky.
6. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Juanich, G. L., Cuevas-Uribe, R. & Mims, S. D. 2013. The culture of paddlefish in reclaimed water disinfected with ozone and/or ultraviolet radiation. KAS 99th Annual Meeting. Kentucky Academy of Science, Morehead, Kentucky.
7. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Juanich, G. L., Cuevas-Uribe, R. & Mims, S. D. 2013. Reuse technology using decommisioned facilities and reclaimed water for hybrid striped bass production. Pages 530. Aquaculture 2013. World Aquaculture Society, Nashville, Tennessee.
8. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Mims, S. & Cuevas-Uribe, R. 2013. Reuse of decommissioned wastewater facilities supplied with reclaimed water for aquaculture. Pages 424. SETAC North America 34th Annual Meeting. Society of Environmental Toxicology and Chemistry, Nashville, Tennessee
9. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Mims, S. D. & Cuevas-Uribe, R. 2013. Contaminant bioaccumulation in paddlefish and hybrid striped bass fingerlings cultured in reclaimed effluent water at a decommissioned wastewater treatment facility. Pages 39. Diversification in Inland Finfish Aquaculture (DIFA II). University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic.
10. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Parrott, T. & Mims, S. D. 2013. A novel business model using decommissioned wastewater treatment tanks for paddlefish fingerling culture and municipal reservoir for growout. Pages 830. Aquaculture 2013. World Aquaculture Society, Nashville, Tennessee.
11. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Squadrito, A., Cuevas-Uribe, R. & Mims, S. D. 2013. Effect of live versus commercially available prepared diets on growth and survival rates of larval paddlefish, Polyodon spathula (Walburn). KAS 99th Annual Meeting. Kentucky Academy of Science, Morehead, Kentucky.
12. Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Squadrito, A., Cuevas-Uribe, R. & Mims, S. D. 2013. Intensive pond production of paddlefish Polyodon spathula in Kentucky. Pages 1048. Aquaculture 2013. World Aquaculture Society, Nashville, Tennessee