Source: AUBURN UNIVERSITY submitted to
BIOMANIPULATION AS A TOOL TO ENHANCE AQUACULTURE THROUGH THE MANAGEMENT OF TOXIC CYANOBACTERIA
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
EXTENDED
Funding Source
Reporting Frequency
Annual
Accession No.
1013884
Grant No.
2017-70007-27132
Project No.
ALA017-1-08011
Proposal No.
2017-04299
Multistate No.
(N/A)
Program Code
AQUA
Project Start Date
Sep 1, 2017
Project End Date
Aug 31, 2020
Grant Year
2017
Project Director
Wilson, A. E.
Recipient Organization
AUBURN UNIVERSITY
108 M. WHITE SMITH HALL
AUBURN,AL 36849
Performing Department
Fisheries, Aquaculture, and Aq
Non Technical Summary
Program and Priority Area: Aquaculture Research; Design of Environmentally and Economically Sustainable Aquaculture Production SystemsRelevance to Program: Blooms of cyanobacteria poison aquaculture ponds around the world. Current management strategies that rely on herbicides or mixing are largely ineffective over the long-term. Building on new preliminary data documenting large effects of zooplankton genotypes that are resistant to toxic cyanobacteria, we propose to use ecological interactions (i.e., trophic cascade theory) as an environmentally and economically sustainable approach for controlling toxic and off-flavor-producing cyanobacteria in commercial catfish aquaculture.Project goal: Promote an industrial paradigm shift towards using foodweb manipulations to control toxin- and off-flavor-producing cyanobacteria to enhance water quality and aquaculture production.Specific objectives: (1) Experimentally determine how water quality in aquaculture ponds is affected by foodweb manipulations, including the use cyanobacteria-tolerant zooplankton, (2) Quantify changes in the quality of commercial catfish exposed to different foodwebs with or without planktivorous fishes, (3) Track aquaculture pond phytoplankton and zooplankton community and trait dynamics, (4) Train students in experimental design, aquaculture, water quality analyses, outreach, and publishing, and (5) Disseminate project findings to aquaculture professionals, extension agents, legislators, and academics through annual workshops, reports, conference presentations, flyers, newsletters, and scientific publications.Approach: Using replicated whole pond experiments, we will determine the effectiveness of using biomanipulation to control toxic and off-flavor producing cyanobacterial blooms in aquaculture ponds. We will also determine the impacts of foodweb manipulations on catfish aquaculture production, quality, and economics. Complementary monitoring and small-scale lab experiments will determine the traits of plankton responsible for the promotion or control of cyanobacterial blooms in aquaculture ponds. Extensive outreach activities will be used to share our findings.
Animal Health Component
0%
Research Effort Categories
Basic
25%
Applied
75%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3073710107040%
3140210107020%
3063710102010%
3063710107010%
3080210119020%
Goals / Objectives
Project goalsShort-term goal: Determine the feasibility of using sustainable foodweb manipulations (i.e., planktivore removal and large-bodied zooplankton addition; trophic cascade theory) to control toxic and off-flavor producing cyanobacteria in hyper-eutrophic catfish aquaculture ponds.Long-term goal: Cause an industrial paradigm shift in freshwater aquaculture towards the use of biomanipulations that promote large-bodied, cyanobacterial-tolerant zooplankton to control toxic cyanobacterial blooms in catfish aquaculture ponds.Project objectivesExperimentally determine how water quality in catfish aquaculture ponds is affected by foodweb manipulations, including (a) removing all planktivorous fishes, (b) adding large-bodied zooplankton (including genotypes that have been documented to be tolerant to toxic cyanobacteria), and/or (c) adding common planktivores (threadfin shad (Dorosoma petenense) and/or tilapia (Oreochromis niloticus)); considered as the "control" given their frequent use in aquaculture).Quantify changes in aquaculture-produced catfish (i.e., feed conversion ratio, survival, growth, fillet color and texture, off-flavors, and cyanobacterial toxins) exposed to different foodwebs with or without planktivorous fishes and/or large-bodied zooplankton.Regularly track aquaculture pond phytoplankton and zooplankton community dynamics, including associated traits, such as plankton diversity, size, and abundance, as well as, zooplankton tolerance to cyanobacteria, through the growing season.Broadly train graduate and undergraduate students in experimental design, aquaculture, water quality sampling and analyses, outreach, networking, and results dissemination.Disseminate project findings to aquaculture farmers, extension agents, and legislators through workshops, newsletters, reports, conference presentations, and journal articles.
Project Methods
The technical aspects of this project involve (1) regular monitoring of 20+ aquaculture ponds at the Auburn University E. W. Shell Fisheries Center and active aquaculture farms in Alabama, (2) analytical water quality analyses, (3) mechanism-based lab experiments, (4) whole pond experiments, and (5) statistical modeling. Aquaculture pond water quality monitoringAt least every month during the first year of the project, our observational efforts will include standard monitoring of 20+ aquaculture ponds at active farms in west Alabama and Auburn University. We will plan to sample biweekly during the primary algal growing season (May-September). Given the wide variability in aquaculture pond use, we expect our monitoring efforts to capture important water quality dynamics within and across ponds. At each pond, a YSI multisensor and Li-Cor light meter will also be used to collect temperature/oxygen/conductivity/pH and light profiles, respectively, from the surface to near the bottom (0.25 m intervals) in the mid-afternoon to minimize the influence of overnight surface aeration. Transparency will be measured with a Secchi disk. Integrated, photic zone water samples will also be collected with a clear plastic tube from the water surface to the depth where 1% of the surface light penetrates. These water samples will be stored in acid-rinsed plastic bottles on ice for later processing. At the lab, each sample will be mixed well and stored in an acid-rinsed plastic bottle for nutrient analyses (total nitrogen, total phosphorus, soluble reactive phosphorus, nitrate), a glass bottle with 1% Lugol's preservative for microscopic phytoplankton enumeration, in a gas chromatograph sample vial to determine concentrations of two common, off-flavor compounds, geosmin and MIB, and collected on glass-fiber filters for chlorophyll, phycocyanin, or algal toxins (microcystin, saxitoxin, and cylindrospermopsin) analyses. Macrozooplankton will be sampled with replicate horizontal hauls of a 0.3 m diameter net (80 μm mesh), preserved in 95% ethanol, and enumerated digitally at 40X. Every month, planktivorous fish diversity and density will be assessed using seines in each monitored aquaculture pond at Auburn University. Given the unnecessary cost and stress on the fish associated with seine hauls, we will collect planktivores when commercial farmers seine their ponds. After collection, planktivorous fish (~10-20/pond) will be euthanized using MS-222 and immediately placed on ice. At the lab, fish gut contents will be analyzed. Laboratory experiments with ambient and cultured phytoplankton and zooplanktonTo determine how variation in important traits that influence plankton interactions and water quality in aquaculture ponds varies seasonally and across ponds, we will conduct two types of mechanistic experiments using natural and cultured plankton during contrasting seasons (spring and fall) from 5+ monitored ponds during the first year of the project. Media for both types of experiments will come directly from the ponds sampled. These experiments will allow us to make general comparisons of mechanisms mediating plankton interactions (e.g., phytoplankton size or toxicity; zooplankton body size or tolerance to toxic food) across sites and time. The PIs have successfully used these approaches in the past and will use D. pulicaria genotypes currently in culture and shown to be either sensitive or tolerant to toxic cyanobacteria based on prior lab and field experiments.Whole pond experimentsIn both project years, we will conduct long-term, whole pond experiments using at least 36, 0.04 ha ponds (6 treatments; 6 replicates/treatment) to determine if biomanipulation can effectively reduce or eliminate cyanobacteria in active aquaculture ponds. We will prepare at least 10 more ponds than needed for each experiment to avoid unforeseen issues that could affect our experimental design.A replicated whole pond experiment will be conducted in each project year using six distinct treatments (Table 1) to evaluate the effect of the current aquaculture management approaches (treatments 2 and 3) versus treatments that include large-bodied Daphnia cultures known to be tolerant or sensitive to toxic cyanobacteria. Over two years, we will conduct two replicated whole pond experiments to confirm the robustness of our expected findings that tolerant Daphnia can control toxic cyanobacteria in aquaculture ponds. Table 1. Whole pond experimental design(1) Ambient (no Daphnia; no planktivorous fish) and catfish(2) Threadfin shad (no Daphnia) and catfish (current management approach)(3) Tilapia (no Daphnia) and catfish (current management approach)(4) Cyanobacteria-tolerant Daphnia pulicaria (no planktivorous fish) and catfish(5) Cyanobacteria-sensitive Daphnia pulicaria (no planktivorous fish) and catfish(6) Planktivorous fish (shad and tilapia), cyanobacteria-tolerant and cyanobacteria-sensitive Daphnia pulicaria, and catfishPrior to each experiment, Daphnia cultures will be maintained outdoors and fed Ankistrodesmus to maximize stocking densities (1/L, sum of all clones). At least three D. pulicaria genotypes will be used per treatment. We will use channel catfish (Ictalurus punctatus) (mean total length of 10 cm) in all treatments and threadfin shad (Dorosoma petenense) and/or tilapia (Oreochromis niloticus) will serve as the planktivorous fish (in relevant treatments) given their frequent use in aquaculture. Fish will be measured and weighed prior to stocking at relevant densities (i.e., 12,500 catfish/ha and 50 kg planktivore/ha). All ponds will be daily fed a floating feed (32% crude protein) based on estimated fish weight (2-3%). Experimental ponds will be sampled every two weeks using standard limnological protocols. Sampling will start at least four weeks prior to the start of each experiment to collect sufficient initial conditions data. Briefly, during each sampling we will measure dissolved oxygen, temperature, pH, and conductivity in situ with a handheld meter and transparency will be measured with a Secchi depth and light meter. Depth-integrated water samples will be collected with a tube sampler and processed in the lab to measure water quality parameters. Zooplankton will be preserved in absolute ethanol and at least 100 D. pulicaria in each Daphnia treatment will be genetically analyzed using four microsatellites that have been shown to discriminate each of the cultured clones when mixed. Each experiment will last at least 6 months to allow for growth of fingerlings. At the end of each experiment, all fish from each pond will be collected, measured, and weighed to determine survival and growth rate. Muscle tissue of three individuals of each fish species per pond will be analyzed monthly for cyanobacterial toxins to determine if cyanotoxin bioaccumulation is a human-health threat. Fish muscle tissue (3/pond collected monthly) and whole water samples will also be analyzed for common cyanobacterial-derived off-flavor compounds, geosmin and 2-methylisoborneol. Planktivorous fish will be collected monthly (3/pond/species) to conduct gut content analysis to confirm that these fish are consuming Daphnia. Flesh color of 3 catfish per replicate per treatment will also be evaluated for irregularities (i.e., yellow fillet). We will also calculate feed conversion ratios (FCR) of catfish in all treatments.

Progress 09/01/18 to 08/31/19

Outputs
Target Audience:This research project is highly applied and relevant to aquaculture farmers, the general public, academics, and natural resource managers given that it directly addresses biotic and abiotic factors that affect water quality in aquaculture ponds. Our large-scale pond monitoring efforts directly connect us with fish farmers with the goal of better understanding how water quality dynamics influence fish health. The basic ecology questions related to biomanipulation should interest ecologists who are interested in using food-web manipulations to influence ecosystem function and community structure. From 1 September 2018 to 31 August 2019 this project has led to five peer-reviewed journal articles (Thurlow et al. 2019, Wadsworth et al. 2019, Chislock et al. 2019, Chislock et al. 2019, Barros et al. 2019), two extension articles (Garner et al. 2019, Buley et al. 2019), four invited seminars by Wilson (Wayne State University, Danube Research Institute Hungary, Florida Atlantic University, Centro de Estudios Ambientales de Cienfuegos Cuba), nine regular conference presentations (World Aquaculture Society, Auburn University's School of Fisheries, Aquaculture, & Aquatic Sciences 2019 Student Symposium, Auburn University This is Research Symposium, Alabama Water Resources Conference), and two extension presentations (2 in Alabama) that reached well over 100 farmers. Wilson co-organized one conference session at the 2019 Association for the Sciences of Limnology and Oceanography (ASLO) in San Juan, Puerto Rico, about undegraduate research with P. Crumrine, G. Hoffmann, D. Fields, S. Rogers, and S. Cohen (Title: Exploring what makes undergraduate research experiences work: evidence from students and mentors). The session was very well attended. Lastly, Wilson and his colleagues, A. Robertson, M. Waters, and D. Hambright, successfully proposed to bring the 10th US Symposium on Harmful Algae planned for November 2019 to Orange Beach, Alabama. This will be a great opportunity to educate the public, aquaculture farmers, agency scientists, and academics about many aspects of algal blooms. Our abstract deadline ends 31 August 2019. We are expecting the conference to be well-attended. I have encouraged our catfish farmer collaborators to join us at the conference. Wilson also participated in a variety of outreach events including (1) participated in a workshop organized by the Alabama Department of Environmental Management to educate state-wide water utilities about risks associated with cyanobacterial blooms, (2) Met with the City of Opelika Water Board to discuss their water quality issues, (3) led discussion about undergraduate research opportunities for College of Sciences and Mathematics and College of Engineering programs targeting incoming under-represented minority freshmen, and (4) organized an outreach event at a local daycare where nine of our NSF REU students shared their research findings with children ranging in age from 2-10. Lastly, Wilson offered to provide an early review of a harmful algal bloom report that was recently released by the Natural Resources Defense Council. Changes/Problems:The only major changes to our project were the start and end dates of our first large pond experiment. We intended to start this experiment early spring 2018 but pond renovations delayed the start of our experiment until late summer. The first experiment got underway in July 2018 and ended February 2019. We have already prepared our 24 ponds and stocked 8 with bluegill collected from our station ponds to give the fish time to equilibrate to the ponds and to affectfoodweb structure. We will plan to start our second experiment this fall once the weather cools a bit. What opportunities for training and professional development has the project provided?The participating graduate and undergraduate students have been involved with many aspects of the project, including experimental pond preparation, experimental design, experiment initiation and sampling, pond monitoring, lab-based water quality analyses, outreach, and the sharing of project results through publications and presentations. Two of the undergraduate students participated in Wilson's NSF Research Experiences for Undergraduates Site in warm-water aquatic ecology and were provided abundant professional development training, such as preparing resumes, identifying graduate programs and advisors, applying for fellowships, and preparing poster and oral presentations. Graduate students participated in weekly lab meetings where Wilson and his students share information and data about their projects. Graduate students are also encouraged to join Wilson in providing ad hoc reviews of manuscripts (after getting approval from journal). All students also participate in lab social events, such as cookouts. How have the results been disseminated to communities of interest?Summary of Extension Activities - 2018-2019 A number of Extension-related activities have been carried out in Year 2 of this project. In 2018, an Extension farmer survey conducted by the Alabama Fish Farming Center (AFFC) revealed a total loss of 214,600 lbs of catfish (214,200 individual catfish) to toxic algae related fish kills.This number is down by more than 30% compared to 2017 (692,000 lbs in reported fish losses due to toxic algae). Previous years have also revealed fluctuations in the numbers of pounds of catfish lost to toxic algae events. In 2016 and 2015, farmers reported losses of 870,000 pounds and 310,000 pounds of catfish, respectively. During 2018-2019, there have been 12 reported or suspected fish kills documented by the Alabama Fish Farming Center in which toxic algae blooms are suspected. In each case, assistance was provided by AFFC personnel in diagnosing the problem either in the lab or with a farm visit to the site of the fish kill. Some of these calls and farm visits were after normal working hours or on weekends. Management recommendations were provided to farmers in consultation with either the fish health diagnostician at the Alabama Fish Farming Center and/or Dr. Wilson. In addition to these officially documented cases, assistance was provided on multiple occasions over the phone or via text to commercial producers even when no fish were brought to the laboratory for direct examination.In addition to diagnostic cases, the AFFC also provided assistance with treatment recommendations to farmers bringing in water samples with heavy blue green algae blooms. At the request of the farmer, additional water samples were shipped from the AFFC to the Wilson Lab for more in depth analyses of algae in the ponds. In 2019, additional efforts are underway to further investigate the impact of toxic algae blooms on Alabama aquaculture farms. Extension funds from this proposal were used to secure a microplate reader for the AFFC. During the 2019 field season, water samples in which there are suspected fish kills caused by toxic algae are being frozen (-20°C) for future analysis of microcystin toxins via an ELISA kit (Abraxis). There are over 1,500 catfish production ponds in west Alabama and this component of the study will provide additional information beyond the 21 ponds that are being routinely sampled during the field season by Dr. Wilson's lab. This ELISA technique is utilized in Dr. Wilson's laboratory on campus for research purposes but the AFFC is exploring its use as an additional tool for the diagnostic laboratory and extension programs in west Alabama. At the end of the field season these samples will be processed to determine whether or not microcystin toxin was present in the water at the time of each documented fish kill. Each water sample is being labeled with a case number assigned from the diagnostic laboratory in order to track data associated with each diagnostic case. Dr. Anita Kelly (recently hired Extension Specialist and Research Professor in Fish Health) is collaborating on this aspect of the project by agreeing to save water samples for disease cases she diagnoses in which toxic algae is suspected as a causative agent. This effort is being carried out for diagnostic cases brought to the AFFC by commercial producers and also cases documented by Dr. Kelly in the Mobile Disease Laboratory (a recreational vehicle equipped with a laboratory used by the AFFC to travel to farms to perform on-farm diagnostic work). In addition to water samples from documented fish kills, additional water samples are also being saved for water quality cases brought in to the Fish Center in which microcystis is believed to be having an impact on consumption of commercial feed in instances of heavy blooms. Farmers have been reporting reduced feed consumption in cases of heavy microcystis blooms, hence, these samples will also be tested for microcystin toxin. To date a total of 47samples have been compiled for this effort and the collection will continue throughout this field season. During the 2019 field season, Dr. Roy's laboratory is providing logistical support to Dr. Wilson's team during pond sampling in west Alabama. Dr. Wilson's team samples 21 ponds in west Alabama typically every 3-4 weeks. On alternating sampling events, Dr. Roy's laboratory has been sampling fish (3-4 fish per pond; either by snagging or hook and line) to obtain fish muscle and liver samples. The samples are either frozen or kept on ice before being transferred to Dr. Wilson's team for transport back to campus for further analyses. Dr. Roy will also provide assistance in securing pertinent information related to treatment and fish kills in the study ponds in west Alabama. Dr. Roy's laboratory is assisting Dr. Wilson's research team by supporting several extension demonstrations on commercial catfish farms in west Alabama designed to evaluate two different strategies to reduce blue green algae in commercial catfish ponds.These demonstrations were initiated the previous year and are being continued this field season. These field demonstrations are assessing the use of hydrogen peroxide as a long-term control measure for blue green algae and the use ofDaphniaas a biological control for blue green algae.To that end, Dr. Roy's laboratory is providing support to one commercial farm that has been using hydrogen peroxide in 17 ponds. In 2018, assistance was also being provided to two inland low salinity shrimp farmers using hydrogen peroxide to control blue green algae (one in Alabama and one in Florida). Currently, there are two commercial catfish farmers that are evaluating the use ofDaphniain two ponds on their farms and comparing them to several control ponds for which traditional management strategies for blue green algae control are being utilized. Additional Extension activities were the inclusion of a project update by the Wilson Lab at the Annual Catfish Update Meeting (approximately 120 attendees) organized by the AFFC and the School of Fisheries, Aquaculture, & Aquatic Sciences at Auburn on December 4, 2018. The AFFC also sponsored an Algae Workshop by Dr. Wilson on January 7, 2019 (16 farmer and hatchery attendees). The workshop was very applied in nature and involved hands on training in the proper use of microscopes and algae identification. Producers were encouraged to bring water samples from their farms. There were several comments from farmers in attendance that due to the applied nature of the workshop it was one of the best Auburn sponsored workshops they had ever attended. An additional workshop is being organized for the Fall of 2019. Dr. Wilson's group published an extension newsletter article in a 2019 issue ofFish Farming News, which is a newsletter published by the Alabama Fish Farming Center. This newsletter was mailed out to 274 farmers and industry stakeholders in addition to being posted online on the website of the Alabama Fish Farming Center. What do you plan to do during the next reporting period to accomplish the goals?We will continue our regular pond monitoring efforts in Auburn and west Alabama throughout 2019. We will also start and complete our second whole pond experiment which we expect to harvest spring 2020. In addition to our field studies, we will add the proposed lab experiments using water collected from the experimental ponds and functioning aquaculture ponds. We will also organize and conduct water quality workshops in west Alabama and Arkansas.

Impacts
What was accomplished under these goals? In our second project year, we made significant progress to achieve our proposal goals and objectives. First, we successfully completed our first whole pond field experiment. We started sampling the 24 ponds on 5 July 2018. After sampling two times, we added 500 catfish to each pond on 26 June 2018. We sampled one more time before we added 0.65 kg of either threadfin shad or bluegill to 6 ponds. On 29 August 2018, we added Daphnia pulicaria (6 different clones) to a plastic mesocosm in each of the remaining 6 ponds. We regularly monitored the pondwater quality until February 2019 when we harvested the catfish. Our fish survival was excellent (89% except for one pond that had a complete die-off after an aerator overheated one night and another had an ichevent during harvesting). However as we harvested the ponds, we realized that our fish treatment effects (bluegill or shad) were small. We found few shad survived the experiment (water may have gotten too cold) and the bluegill, although alive, had just spawned in several of the bluegill ponds and their densities were fairly low. We based our stocking densities of bluegill on past studies, but have decided to increase stocking density 4x in the second experiment. We recently confirmed that our 8 bluegill ponds have healthy, dense populations of bluegill. Our treatments effects were small. Most of the trends were consistent across treatments. We are expecting larger treatment effects this time. We would like to attachslides of our current data. In addition to our whole pond experiment, we have been monitoring 21 catfish production ponds in west Alabama (across 5 farms; 3x thus far) and 10 catfish production ponds at the Shell Fisheries Center (3x thus far). Water quality in the ponds varies a lot over space and time. West Alabama ponds tend to be more productive and have higher toxin concentrations than Auburn ponds. Although we do see trends in higher phosphorus and nitrogen during the growing season (likely as a result of feed). We don't see clear correlations between the algal toxin, microcystin, and phytoplankton (measured as chlorophyll), likely because cyanobacteria (measured as phycocyanin) doesn't get abundant until the early summer. Two of the farmers have tried to keep out planktivorous fishes when filling their ponds (Clemmer 1 and Miller 1, 20, 29). We have found that several of the ponds with no planktivorous fish provide good habitat for abundant large-bodied zooplankton that have caused large reductions in algal abundance. One side effect of low phytoplankton abundance is that dissolved nutrients, namely nitrate, nitrite, and ammonia, get very high since limited algae are available to use the nutrients. Cyanobacteria and microcystin increase dramatically in the summer. This pattern has been consistent in 2018 and 2019. We would like to attachslides of our current data. Roy is currently gathering information from the five collaborating catfish farms so that we can see if any patterns in the water quality data align with any significant events, such as fish kills, draining, and chemical applications. We have not started the lab bioassays testing for trait changes in phytoplankton and zooplankton over time, but I have recruited a new MS student, Angelea Belfiore, who will begin this component of our project.Angelea Belfiore joined us a technician in July 2019 and recently started her MS this month. Because off-flavor has been fairly low in our aquaculture ponds, we didn't finalize our method for measuring taste and odor compounds in fish tissue using microwave extraction. Our recovery rates (~60%) are near published rates but we wanted to optimize our method to increase the recovery rates before we start processing tissue samples. Wilson recruited two new graduate students, Matt Gladfelter (MS) and Hengxing Tang (PhD), who started working with us 1 August 2018. Matt is doing very well and I am optimistic that I can keep Matt around for a PhD. Hengxing struggled in the lab, and ultimately left Auburn University in December 2018. In addition, 9 undergraduate students have participated in this project to date. Two of these undergraduates, Catie Adams and Kathryn Cruz, conducted research relevant to aquaculture systems but were primarily funded through my NSF REU grant. Catie tested several algaecides of companies that I met after attending the WAS meeting in New Orleans. Her data from lab and field experiments are very interesting and show how little time algaecide treatments last. We are still finalizing some data analyses before we write this paper up for publication. Kathryn was developing a new method to use a drone to monitor water quality, especially cyanobacterial abundance. Our lab is excellent right now, but given the undergraduates' work schedules, I might recruit more undergraduates to join our team this semester. One major activity that was not proposed is the use of hydrogen peroxide to control algal blooms. One of Wilson's postdocs, Dr. Zhen Yang, recently published results from lab and field experiments clearly showing how hydrogen peroxide (~7 mg/L) can be used to control cyanobacteria. We have shared these results with many farmers in west Alabama and have learned that many farms are using this method to control algae in their ponds. One farmer recently told Wilson that his utility bills have been significantly lower after he started the peroxide treatments. Riley Buley, a PhD student, in Wilson's lab is developing his dissertation research around using algaecides and ecology to control algal blooms in aquaculture ponds. There has been a lot of interest by vendors to have us test their products. Catie and Riley have conducted a variety of experiments that show how the effectiveness of the treatments vary but also highlight the disconnect between lab and field results. Ultimately, we need to do more experiments in the field in functioning aquaculture ponds.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Wilson, A. E. 2018. Water quality trends in catfish production ponds. Annual catfish update meeting, Auburn University, School of Fisheries, Aquaculture, and Aquatic Sciences. Demopolis, Alabama.
  • Type: Other Status: Published Year Published: 2019 Citation: Wilson, A. E., Roy, Luke, L. A. 2019. Phytoplankton enumeration water quality workshop, Alabama Fish Farming Center. Greensboro, Alabama
  • Type: Other Status: Published Year Published: 2019 Citation: Wilson, A. E. 2019. Consequences of nutrient enrichment and herbivore adaptation for phytoplankton in lakes. Florida Atlantic University, Department of Biology, Boca Raton, Florida. (keynote lecture)
  • Type: Other Status: Published Year Published: 2019 Citation: Wilson, A. E. 2019. Consequences of nutrient enrichment and herbivore adaptation for phytoplankton in lakes. Wayne State University, Department of Biological Sciences. Detroit, Michigan. (invited lecture)
  • Type: Other Status: Published Year Published: 2018 Citation: Wilson, A. E. 2018. Consequences of nutrient enrichment and herbivore adaptation for phytoplankton in lakes. Danube Research Institute, MTA Centre for Ecological Research. Budapest, Hungary. (invited lecture)
  • Type: Other Status: Published Year Published: 2018 Citation: Wilson, A. E. 2018. Do harmful algal blooms threaten freshwater resources in the southeastern U.S.? Centro de Estudios Ambientales de Cienfuegos, NSF PIRE workshop. Cienfuegoes, Cuba. (invited lecture)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Johnston, L. and A. E. Wilson. 2019. Exploring the removal of excess nutrients and heavy metals from polluted waterbodies through the use of macrophytes and microorganisms. This is Research: Student symposium. Auburn, Alabama. (poster)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Buley, R. P., M. F. Gladfelter, and A. E. Wilson. 2019. Integrated methods combat cyanobacterial blooms and off-flavors in the aquaculture and drinking water industries. This is Research: Student symposium. Auburn, Alabama.
  • Type: Other Status: Published Year Published: 2019 Citation: Garner, B., C. Adams, R. P. Buley, and A. E. Wilson. Aquaculture 2019: Auburn students perspective. World Aquaculture 50(2):19.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Buley, R., Z. Yang, M. F. Gladfelter, and A. E. Wilson. 2019. Controlling blue-green algal blooms in aquaculture pond using hydrogen peroxide. Fish Farming News 2019(1):3-5.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Barros, M. U.G., A. E. Wilson, J. I. R. Leit�o, S. P. Pereira, R. P. Buley, E. G. Fernandez-Figueroa, and J. C. Neto. 2019. Environmental factors associated with toxic cyanobacterial blooms across 20 drinking water reservoirs in a semi-arid region of Brazil. Harmful Algae 86:128-137.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Chislock, M. F., O. Sarnelle, L. M. Jernigan, V. R. Anderson, A. Abebe, and A. E. Wilson. 2019. Consumer adaptation mediates top-down regulation across a productivity gradient. Oecologia 190:195-205.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Chislock, M. F., R. B. Kaul, K. A. Durham, O. Sarnelle, and A. E. Wilson. 2019. Eutrophication mediates rapid clonal evolution in Daphnia pulicaria. Freshwater Biology 64:1275-1283.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Chislock, M., L. Denecke, M. S. Sepulveda, O. Sarnelle, and A. E. Wilson. 2019. Intraspecific variation in lake Daphnia affects response of algae and a cyanotoxin to fertilization. International Association of Great Lakes Research. The College of Brockport, State University of New York. Brockport, New York.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Davenport, Z. and A. E. Wilson. 2018. Can nitrogen to phosphorus ratios be used to improve the quality of drinking water reservoirs? Gallaudet University Undergraduate Research Symposium. Gallaudet University. Washington, D.C. (poster)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Lamb, A. N., S. E. Leinbach, M. N. Water, and A. E. Wilson. 2018. Linking photosynthetic pigments and taxonomy in cyanobacteria blooms: implications for water quality management in the Southeast United States. 2018 College of Agriculture Student Poster Showcase. Auburn University, Auburn, Alabama. (poster) (2nd place student poster presentation)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Buley, R. P. and A. E. Wilson. 2018. Understanding microcystin patterns in freshwater ecosystems: a meta-analysis. 2018 College of Agriculture Student Poster Showcase. Auburn University, Auburn, Alabama. (poster)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Lamb, A. N., S. E. Leinbach, M. N. Water, and A. E. Wilson. 2018. Linking photosynthetic pigments and taxonomy in cyanobacteria blooms: implications for water quality management in the Southeast United States. Alabama Water Resources Conference, Orange Beach, Alabama (poster) (1st place student poster presentation)
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Thurlow, C. M., M. A. Williams, C. M. Thurlow, A. Carrias, C. Ran, M. Newman, J. Tweedie, E. Allison, L. N. Jescovitch, A. E. Wilson, J. Terhune, and M. R. Liles. 2019. Bacillus velezensis AP193 exerts probiotic effects in channel catfish (Ictalurus punctatus) and reduces aquaculture pond eutrophication. Aquaculture 503:347-356.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Wadsworth, P., A. E. Wilson, and W. C. Walton. 2019. A meta-analysis of growth rate in diploid and triploid oysters. Aquaculture 499:9-16.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Wilson, A. E., Yang, Z., R. P. Buley, E. Fernandez-Figueroa, M. Barros, and S. Rajendran. 2019. Hydrogen peroxide treatment promotes chlorophytes over toxic cyanobacteria in a hyper-eutrophic aquaculture pond. World Aquaculture Society. New Orleans, Louisiana.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Buley, R. P. and A. E. Wilson. 2019. Understanding microcystin patterns in freshwater ecosystems: a meta-analysis. World Aquaculture Society. New Orleans, Louisiana.


Progress 09/01/17 to 08/31/18

Outputs
Target Audience:This research project is highly applied and relevant to aquaculture farmers, the general public, academics, and natural resource managers given that it directly addresses biotic and abiotic factors that affect water quality in aquaculture ponds. Our large-scale pond monitoring efforts directly connect us with fish farmers with the goal of better understanding how water quality dynamics influence fish health. The basic ecology questions related to biomanipulation should interest ecologists who are interested in using food-web manipulations to influence ecosystem function and community structure. From 1 September 2017 to 31 September 2018, this project has led to three peer-reviewed journal articles (Wilson et al. 2018, Yang et al. 2018, and Baker et al. 2018), two manuscripts in revision (Thurlow et al., Wadsworth et al.), two extension articles (Wilson and Fernandez-Figueroa 2018, Wilson et al. 2018), two regular conference presentations (Association for the Sciences of Limnology and Oceanography annual conference; Auburn University's School of Fisheries, Aquaculture, & Aquatic Sciences 2018 Student Symposium), and five extension presentations (4 in Alabama and 1 in Arkansas) that reached well over 100 farmers. Wilson co-organized two conference sessions at the 2018 Association for the Sciences of Limnology and Oceanography (ASLO) in Victoria, British Columbia, about harmful algal blooms with B. Brooks, J. Steevens, and M. Howard (Title: Crossing disciplinary boundaries across the freshwater-marine continuum to advance the understanding of harmful algal blooms (HABs)) and T. Harris, D. Van De Waal, F. Pick, P. Leavitt, and S. Wood (Title: Cyanobacterial and algal metabolites: occurrence, ecology, prediction, and management). Both sessions were attended. Lastly, Wilson and his colleagues, A. Robertson, M. Waters, and D. Hambright, successfully proposed to bring the 10th US Symposium on Harmful Algae planned for November 2019 to Orange Beach, Alabama. This will be a great opportunity to educate the public, aquaculture farmers, agency scientists, and academics about many aspects of algal blooms. We are in the final stages of choosing the meeting location and dates. Wilson also participated in a variety of outreach events including (1) participated in a workshop organized by the Alabama Department of Environmental Management to educate state-wide water utilities about risks associated with cyanobacterial blooms, (2) participated in an outreach event led by the Auburn University Writing Center to support Auburn University faculty how to write successful NSF CAREER proposals, (3) led discussion about undergraduate research opportunities for College of Sciences and Mathematics and College of Engineering programs targeting incoming under-represented minority freshmen, (4) participated in the 2018 Lee County Water Festival to teach hundreds of 4th graders about the importance of aquifers, (5) participated in outreach event led by the College of Agriculture's USDA-funded Science in Agriculture, Food and the Environment (S.A.F.E.) program to teach under-represented middle school students about water quality, and (6) organized an outreach event at a local daycare where nine of our NSF REU students shared their research findings with children ranging in age from 2-10. Changes/Problems:The only major changes to our project werethe start date of our first large pond experiment and a reduction in available ponds. We intended to start this experiment early spring but pond renovations delayed the start of our experiment until late summer. We will plan to start our second experiment on time in 2019. We originally had 36 ponds renovated for our experiment but learned while preparing the ponds that one block(n=12) of ponds was more leaky than we were comfortable using. So, we removed that block from our experimental design and reduced our experimental design to 4 treatments (6 ponds per treatment). What opportunities for training and professional development has the project provided?The participating graduate and undergraduate students have been involved with many aspects of the project, including experimental pond preparation, experimental design, experiment initiation and sampling, pond monitoring, lab-based water quality analyses, outreach, and the sharing of project results through publications and presentations. Two of the undergraduate students participated in Wilson's NSF Research Experiences for Undergraduates Site in warm-water aquatic ecology and were provided abundant professional development training, such as preparing resumes, identifying graduate programs and advisors, applying for fellowships, and preparing poster and oral presentations. Graduate students participated in weekly lab meetings where Wilson and his students share information and data about their projects. Graduate students are also encouraged to join Wilson in providing ad hoc reviews of manuscripts (after getting approval from journal). All students also participate in lab social events, such as cookouts. How have the results been disseminated to communities of interest?Summary of Extension Activities A number of Extension-related activities have been carried out in Year 1. In December 2017 and January/February 2018, chemical residue avoidance training was carried out by Dr. Roy (Co-PI) to educate farmers on the dangers and risks of over treatment of chemicals utilized in catfish production ponds, including chemicals used to control toxic blue green algae and off-flavor. This training was part of a large-scale effort coordinated by the USDA Food Safety and Inspection Service Catfish Committee and the Catfish Farmers of America Residue Avoidance Training Committee to provide training to stakeholders to reduce the risk of catfish processing plants failing USDA FSIS residue inspections. Dr. Roy is a member of both committees and helped develop the training materials. In 2017 there were two domestic recalls from U.S. processing plants that subsequently led to the development and implementation of this training program. Two 90-minute training sessions were organized and carried out by Dr. Roy in Alabama. The first training session was carried out at the Annual Catfish Update meeting in Demopolis sponsored by the School of Fisheries, Aquaculture & Aquatic Sciences at Auburn University and the second was held at the Alabama Fish Farming Center in Greensboro. At the request of Arkansas Cooperative Extension, Dr. Roy also traveled to assist with the training (90 minute session) at the Catfish Farmers of Arkansas annual meeting. In total, this training reached 97.8% (82.1% in Demopolis; 15.7% in Greensboro) of the farmed water acres of catfish in the state of Alabama. In 2017, a farmer survey conducted by the Alabama Fish Farming Center revealed a total loss of 692,000 lbs of catfish to toxic algae related fish kills. This number is down by more than 20% compared to 2016 (886,000 lbs in reported fish losses due to toxic algae). In 2018, there have been six reported fish kills documented by the Alabama Fish Farming Center that were linked to toxic algae blooms. In each case, Dr. Roy's team provided assistance in diagnosing the problem either in the lab or with a farm visit to the site of the fish kill. Management recommendations were provided to farmers in consultation with either the fish health diagnostician at the Alabama Fish Farming Center and/or Dr. Wilson. In addition to these officially documented cases, assistance was provided on multiple occasions over the phone to commercial producers even when no fish were brought to the laboratory for direct examination. Dr. Roy's laboratory is assisting Dr. Wilson's research team by supporting several extension demonstrations on commercial catfish farms in west Alabama designed to evaluate two different strategies to reduce blue green algae in commercial catfish ponds. These demonstrations are assessing the use of hydrogen peroxide as a long-term control measure for blue green algae and the use ofDaphniaas a biological control for blue green algae. To that end, Dr. Roy's laboratory is providing support to one commercial farm that has been using hydrogen peroxide in 17 ponds. Assistance is also being provided to two inland low salinity shrimp farmers using hydrogen peroxide to control blue green algae (one in Alabama and one in Florida).Currently, there are two commercial catfish farmers that are evaluating the use ofDaphniain two ponds on their farms and comparing them to several control ponds for which traditional management strategies for blue green algae control are being utilized. In total, Dr. Roy's team has made multiple farm visits to each extension demonstration site in year 1 of this project to provide advice and treatment recommendations. Dr. Wilson's group published an extension newsletter article in a 2018 issue ofFish Farming News, which is a newsletter published by the Alabama Fish Farming Center. This newsletter was mailed out to 274 farmers and industry stakeholders in addition to being posted online on the website of the Alabama Fish Farming Center. The alternative management strategies being evaluated by Dr. Wilson's team is generating quite a bit of buzz and excitement among commercial aquaculture producers in west Alabama. What do you plan to do during the next reporting period to accomplish the goals?We will continue our regular pond monitoring efforts in Auburn and west Alabama. We will also complete our first whole pond experiment and then prepare the ponds for our second whole pond experiment to begin early spring 2019. In addition to our field studies, we will add the proposed lab experiments using water collected from the experimental ponds and functioning aquaculture ponds. We will also organize and conduct water quality workshops in west Alabama and Arkansas.

Impacts
What was accomplished under these goals? In our first project year, we made significant progress to achieve our proposal goals and objectives. First, we worked with our facilities crew to renovate thirty-six 0.1 acre ponds to prepare for the whole pond experiment. After filling the 36 ponds, we realized that one block of ponds (n=12) were extra leaky so this set was removed from our experimental design. We are currently using 24 ponds for our experiment. We started sampling on 5 July 2018. After sampling two times, we added 500 catfish to each pond on 26 June 2018. We sampled one more time before we added 0.65 kg of either threadfin shad or bluegill to 6 ponds. On 29 August 2018, we added Daphnia pulicaria (6 different clones) to a plastic mesocosm in each of the remaining 6 ponds. After allowing the Daphnia some time to increase in abundance, we will dump the cans into the ponds. We will continue monitoring the water quality of the 24 ponds until we remove the catfish later this fall. In addition to our whole pond experiment, we have been monitoring 21 catfish production ponds in west Alabama (across 5 farms; 3x thus far) and 10 catfish production ponds at the Shell Fisheries Center (3x thus far). We have not started the lab bioassays testing for trait changes in phytoplankton and zooplankton over time but plan to get the new graduate students working on these analyses soon. We have developed several analytical techniques, including nitrate, nitrite, and total ammonia nitrogen analysis. We are near finalizing our new method for measuring taste and odor compounds in fish tissue using microwave extraction. Our recovery rates (~60%) are near published rates but we are trying to optimize our method to increase the recovery rates before we start processing tissue samples. Wilson recruited two excellent, new graduate students, Matt Gladfelter (MS) and Hengxing Tang (PhD), who started working with us 1 August 2018. In addition, 18 undergraduate students have participated in this project to date, and we have recently recruited more undergraduates to join our team this semester. One major activity that was not proposed is the use of hydrogen peroxide to control algal blooms. One of Wilson's postdocs, Dr. Zhen Yang, recently published results from lab and field experiments clearly showing how hydrogen peroxide (~7 mg/L) can be used to control cyanobacteria. We have shared these results with many farmers in west Alabama and have learned that many farms are using this method to control algae in their ponds. One farmer recently told Wilson that his utility bills have been significantly lower after he started the peroxide treatments.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Hemstreet B, Roy LA, Whitis GN. 2018. 2017 Activities of the Alabama Fish Farming Center. Alabama Farmer Federation Commodity Organizational Meeting. Montgomery, Alabama. Feb. 8.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Roy LA. 2018. Residue training for the U.S. catfish industry. Alabama Fish Farming Center, Greensboro, Alabama. Feb. 2. (15 attendees)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Dorman L, Roy LA. 2018. Residue training for the U.S. catfish industry. Catfish Farmers of Arkansas, Hot Springs, Arkansas. Jan. 11. (12 attendees)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Roy LA, Avery J. 2017. Residue training for the U.S. catfish industry. Annual Catfish Update Meeting, Demopolis, Alabama. Dec. 6 (140 attendees)
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Wilson A, Fernandez E, Buley R, Roy LA. 2018. Is sustainable control of toxic algae a reality for catfish aquaculture? Fish Farming News. 2018(1):7-8.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Yang, Z., R. P. Buley, E. G. Fernandez-Figueroa, M. U.G. Barros, S. Rajendran, and A. E. Wilson. 2018. Hydrogen peroxide treatment promotes chlorophytes over toxic cyanobacteria in a hyper-eutrophic aquaculture pond. Environmental Pollution 240:590-598.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Baker, B. C., A. E. Wilson, and J. T. Scott. 2018. Phytoplankton N2 fixation efficiency and its effect on harmful algal blooms. Freshwater Science 37(2):264-275.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Wilson, A. E. and E. G. Fernandez-Figueroa. 2018. Preparing water resource managers to identify and measure toxic cyanobacteria. Streamlines Spring 2018:18.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Wilson, A. E., M. F. Chislock, Z. Yang, M. U.G. Barros, and J. F. Roberts. 2018. Pond bank access as an approach for managing toxic cyanobacteria in beef cattle pasture drinking water ponds. Environmental Monitoring and Assessment 190:247.
  • Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Thurlow, C. M., M. A. Williams, C. M. Thurlow, A. Carrias, C. Ran, M. Newman, J. Tweedie, E. Allison#, L. N. Jescovitch, A. E. Wilson, J. Terhune, and M. R. Liles. Accepted pending revisions. Bacillus velezensis AP193 exerts probiotic effects in channel catfish (Ictalurus punctatus) and reduces aquaculture pond eutrophication. Aquaculture.
  • Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Wadsworth, P., A. E. Wilson, and W. C. Walton. Accepted pending revisions. A meta-analysis of growth rate in diploid and triploid oysters. Aquaculture.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Wilson, A. E. 2017. Controlling toxic algal blooms in catfish aquaculture ponds. Annual catfish update meeting, Auburn University, School of Fisheries, Aquaculture, and Aquatic Sciences. Demopolis, Alabama.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Buley, R. P., Z. Yang, and A. E. Wilson. 2018. Integrated methods combat cyanobacterial blooms in aquaculture. Auburn Universitys School of Fisheries, Aquaculture, & Aquatic Sciences 2018 Student Symposium. Auburn, Alabama.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Fernandez-Figueroa, E. and A. E. Wilson. 2018. Effects of environmental factors on taste and odor compound production. Association for the Sciences of Limnology and Oceanography. Victoria, British Columbia.