Alleviating Water Quality Impacts of Animal Waste through Mycoremediation and Mycofiltration

ALLEVIATING WATER QUALITY IMPACTS OF ANIMAL WASTE THROUGH MYCOREMEDIATION AND MYCOFILTRATION

Sponsoring Institution

National Institute of Food and Agriculture

Project Status

TERMINATED

Funding Source

HATCH

Reporting Frequency

Annual

Accession No.

0212830

Grant No.

(N/A)

Project No.

ALA02-015

Proposal No.

(N/A)

Multistate No.

(N/A)

Program Code

(N/A)

Project Start Date

Oct 1, 2007

Project End Date

Sep 30, 2010

Grant Year

(N/A)

Project Director
Srivastava, P.

Recipient Organization
AUBURN UNIVERSITY
108 M. WHITE SMITH HALL
AUBURN,AL 36849

Performing Department
BIOSYSTEMS ENGINEERING

Non Technical Summary
Nutrients, pathogens, and other trace contaminants in farm animal waste continue to threaten sustainability of water resources of major animal producing states. To reduce threat to water resources from contamination, innovative approaches that (a) do not generate secondary waste streams and (b) add value to wastes are critically needed. The overarching goal of our proposed project is to reduce nutrients and pathogens from animal waste through a novel approach that utilizes remediation/filtration properties of mushrooms (higher macrofungi). The specific objectives are to: (1) Develop procedures to grow selected fungal species on autoclaved and raw wood chips, broiler litter, and solids separated from dairy waste; (2) Conduct laboratory experiments to quantify effectiveness of these fungal species in reducing fecal coliform (Escherichia coli), nitrogen, and phosphorous from broiler litter and dairy wastes; (3) Conduct pilot-scale tests and develop guidelines for low-cost mycoremediation/mycofiltration systems; and (4) coordinate and conduct educational programs for Extension Educators to introduce them to mycoremediation/mycofiltration, and to disseminate the findings of this study. These objectives will be achieved through laboratory experiments, pilot-scale testing, and appropriate extension activities. This innovative approach promises to not only treat farm animal waste but also add value to it, thus providing a sustainable solution.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

100%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
112	0210	1102	30%
112	0210	2020	70%

Knowledge Area
112 - Watershed Protection and Management;

Subject Of Investigation
0210 - Water resources;

Field Of Science
2020 - Engineering; 1102 - Mycology;

Keywords

Goals / Objectives
The overarching goal of our proposed project is to reduce nutrients and pathogens from animal waste through a novel approach that utilizes remediation/filtration properties of mushrooms (higher macrofungi). The specific objectives are to: (1) Develop procedures to grow selected fungal species on autoclaved and raw wood chips, broiler litter, and solids separated from dairy waste; (2) Conduct laboratory experiments to quantify effectiveness of these fungal species in reducing fecal coliform (Escherichia coli), nitrogen, and phosphorous from broiler litter and dairy wastes; (3) Conduct pilot-scale tests and develop guidelines for low-cost mycoremediation/mycofiltration systems; and (4) coordinate and conduct educational programs for Extension Educators to introduce them to mycoremediation/mycofiltration, and to disseminate the findings of this study.

Project Methods
The first objective will be to develop methods to grow the three selected fungal species on wood chips, dairy solids, and broiler litter. Cultures of the selected native mushrooms will be collected from selected wooded locations in Lee and Macon Counties of Alabama, and the gourmet mushroom species will be purchased. These basidiomycetes will be cultivated and propagated on standardized culture media in lab using sterile techniques. Then, mycelium of each species will be transferred to individual substrates composed of each waste treatment. Waste treatments will include sterilized and natural (non-sterilized): 1) wood chips; 2) broiler litter; and 3) dairy solids. Laboratory experiments will be conducted to quantify effectiveness of selected fungal species in reducing fecal coliform (E. coli), nitrogen, and phosphorous from broiler litter and dairy wastes. Selected cultures of the native mushrooms and the gourmet mushroom species will be inoculated onto natural (non-sterilized) broiler litter and dairy solids. Raw waste will be characterized before the start of the experiments. While the wastes are being inoculated with selected mushrooms, it will be sampled, in triplicate, every week to quantify changes in forms of N and P, and counts of E. Coli. Laboratory experiments will also be conducted to quantify effectiveness of selected fungal species in reducing E. coli, nitrogen, and phosphorous from dairy lagoon supernatant. The results from the laboratory experiments will be used to develop pilot-scale systems at E.V. Smith Research Center. These pilot-scale systems will allow us to determine the applicability of mycoremediation/mycofiltration systems in a real-world scenario. One pilot-scale system will be used for remediation of broiler litter and one for filtration of dairy supernatant. Year 1 and Year 2 results will be submitted for publication in refereed journals, including the Agronomy Journal, Environmental Science and Technology, and Journal of Environmental Quality, and presented at locally relevant professional conferences and seminars, such as the ALFA International Poultry Expo, EVSRC Dairy Field Day, and the Sunbelt Expo. In addition to presenting results at conferences and seminars, regional workshops for Extension Specialists, County Extension Agents, Farmers, and other state and private employees with relevance to the wastewater treatment will be organized.

Progress 10/01/07 to 09/30/10

Outputs
OUTPUTS: OUTPUTS: (1) This project was initially started as a three-year project, but received only one year of funding. Because of which, after one year of research, this project was put on-hold. (2) We continue to seek funding for this project from competitive sources. For example, we submitted a pre-proposal to Southern Region Sustainable Agriculture Research and Extension (SARE) program and were invited to submit a full proposal. We submitted the full-proposal in November of 2010 and are currently waiting on the outcome. (3) As reported after one year of project, three strains of mushrooms were tested (Stropharia rugosa annulata, Pleurotus ostreatus and Trametes versicolor) to determine the best cultivation choice for Alabama farmers. The fungi strains were grown on both woodchip and straw autoclaved substrates at three levels of poultry manure concentrations (30:1 C:N, 15:1 C:N, and 10:1 C:N). P. ostreatus substrate and fruit samples collected throughout the growing season are in the process of being analyzed to determine the concentration of nutrients made available for plant uptake, and the level of heavy metal hyperaccumulation in the fungi. Analyses include pH, electrical conductivity, Moisture content, Al, Ca, Cu, Fe, K, Mg, Mn, Na, P, Zn, NH4-N, NO3-N, PO4-P, %N, % C, and ash content. Preliminary results were at the Sand Mountain Research and Extension Center's (SMREC) Poultry Producer Extension Field Day in Crossville, Alabama in October 2007. Attendance included Alabama Poultry Producers, County and Regional Extension Agents, Researchers and the SMREC staff. Farmers in attendance were very receptive to the idea of producing livestock feed and other value added products from materials available on-farm, with little to no additional cost. Presentation was made to Heifer International representatives at the Heifer Ranch near Little Rock, AR in April 2008. Interest was shown in applying our concept in Peru, Ecuador, and Mexico. Presentation was also made to a Food and Agriculture Organization (FAO) representative in Little Rock, AR in April 2008. FAO is interested in the results of the project and has been kept up to date on the project via email and telephone correspondence. The research associate working on the project attended the Fungi Perfecti's mycoremediation workshop. Mycoremediation aspects covered in the workshop included, strain selection, environmental influences on cultivation and remediative efficacy, specific remediative properties of a number of fungi strains, including P. ostreatus. Research Associate also attended the Mycoremediation section of the Sustainable Agriculture Working Group (SAWG) conference. PARTICIPANTS: Jayme Oates, Research Associate, Fisheries and Allied Aquaculture, Auburn University, Auburn, AL. Puneet Srivastava, Associate Processor, Biosystems Engineering, Auburn University, Auburn, AL. (PD). Kathy S. Lawrence, Associate Professor, Entomology and Plant Pathology, Auburn University, Auburn, AL. (Co-PD). The project provided training to two graduate students. TARGET AUDIENCES: Poultry Producers, Mushroom Growers, Soil and Water Conservation Agencies, State Extension Services. The effort will help better manage broiler litter, reduce threat of soil and water contamination, and generated value added products from poultry litter (e.g., mushroom fruits and mushroom compost). PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
(1) Out of the three strains tested, Pleurotus ostreatus was determined to be the best cultivation choice for Alabama farmers due to its fast and hardy growth habits, and tolerance to extreme environmental conditions (temperature, moisture, light, CO2 levels). (2) All three fungi grew successfully on both woodchip and straw autoclaved substrates at the three levels of poultry manure concentrations (30:1 C:N, 15:1 C:N, and 10:1 C:N). However, growth slowed with increasing nitrogen concentrations, and subsequent mushroom fruiting on the highest nitrogen substrate occurred 1-4 weeks after the substrates containing lower nitrogen concentrations.

Publications

No publications reported this period

Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: (1) This project was initially started as a three-year project, but received only one year of funding. Because of which, after one year of research, this project was put on-hold. (2) We continue to seek funding for this project from competitive sources. For example, we submitted a pre-proposal to Southern Region Sustainable Agriculture Research and Extension (SARE) program and were invited to submit a full proposal. We submitted the full-proposal in November of 2009 and are currently waiting on the outcome. (3) As reported after one year of project, three strains of mushrooms were tested (Stropharia rugosa annulata, Pleurotus ostreatus and Trametes versicolor) to determine the best cultivation choice for Alabama farmers. The fungi strains were grown on both woodchip and straw autoclaved substrates at three levels of poultry manure concentrations (30:1 C:N, 15:1 C:N, and 10:1 C:N). P. ostreatus substrate and fruit samples collected throughout the growing season are in the process of being analyzed to determine the concentration of nutrients made available for plant uptake, and the level of heavy metal hyperaccumulation in the fungi. Analyses include pH, electrical conductivity, Moisture content, Al, Ca, Cu, Fe, K, Mg, Mn, Na, P, Zn, NH4-N, NO3-N, PO4-P, %N, % C, and ash content. Preliminary results were at the Sand Mountain Research and Extension Center's (SMREC) Poultry Producer Extension Field Day in Crossville, Alabama in October 2007. Attendance included Alabama Poultry Producers, County and Regional Extension Agents, Researchers and the SMREC staff. Farmers in attendance were very receptive to the idea of producing livestock feed and other value added products from materials available on-farm, with little to no additional cost. Presentation was made to Heifer International representatives at the Heifer Ranch near Little Rock, AR in April 2008. Interest was shown in applying our concept in Peru, Ecuador, and Mexico. Presentation was also made to a Food and Agriculture Organization (FAO) representative in Little Rock, AR in April 2008. FAO is interested in the results of the project and has been kept up to date on the project via email and telephone correspondence. The research associate working on the project attended the Fungi Perfecti's mycoremediation workshop. Mycoremediation aspects covered in the workshop included, strain selection, environmental influences on cultivation and remediative efficacy, specific remediative properties of a number of fungi strains, including P. ostreatus. Research Associate also attended the Mycoremediation section of the Sustainable Agriculture Working Group (SAWG) conference. PARTICIPANTS: Jayme Oates, Research Associate, Fisheries and Allied Aquaculture, Auburn University, Auburn, AL. Puneet Srivastava, Associate Processor, Biosystems Engineering, Auburn University, Auburn, AL. (PD). Kathy S. Lawrence, Associate Professor, Entomology and Plant Pathology, Auburn University, Auburn, AL. (Co-PD). The project provided training to two graduate students. TARGET AUDIENCES: Poultry Producers, Mushroom Growers, Soil and Water Conservation Agencies, State Extension Services. The effort will help better manage broiler litter, reduce threat of soil and water contamination, and generated value added products from poultry litter (e.g., mushroom fruits and mushroom compost). PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
(1) Out of the three strains tested, Pleurotus ostreatus was determined to be the best cultivation choice for Alabama farmers due to its fast and hardy growth habits, and tolerance to extreme environmental conditions (temperature, moisture, light, CO2 levels). (2) All three fungi grew successfully on both woodchip and straw autoclaved substrates at the three levels of poultry manure concentrations (30:1 C:N, 15:1 C:N, and 10:1 C:N). However, growth slowed with increasing nitrogen concentrations, and subsequent mushroom fruiting on the highest nitrogen substrate occurred 1-4 weeks after the substrates containing lower nitrogen concentrations.

Publications

No publications reported this period

Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: (1) Three strains of mushrooms were tested (Stropharia rugosa annulata, Pleurotus ostreatus and Trametes versicolor) to determine the best cultivation choice for Alabama farmers. The fungi strains were grown on both woodchip and straw autoclaved substrates at three levels of poultry manure concentrations (30:1 C:N, 15:1 C:N, and 10:1 C:N). (2) P. ostreatus substrate and fruit samples collected throughout the growing season are in the process of being analyzed to determine the concentration of nutrients made available for plant uptake, and the level of heavy metal hyperaccumulation in the fungi. Analyses include pH, electrical conductivity, Moisture content, Al, Ca, Cu, Fe, K, Mg, Mn, Na, P, Zn, NH4-N, NO3-N, PO4-P, %N, % C, and ash content. (3) P. ostreatus substrate and fruit samples collected over the duration of the growing season are also in the process of being analyzed to determine the remediation capacity of the fungi on anthropogenic substances including antibiotics, steroids, and pesticides commonly used in conventional commercial poultry production. (4) Preliminary results were at the Sand Mountain Research and Extension Center's (SMREC) Poultry Producer Extension Field Day in Crossville, Alabama in October 2007. Attendance included Alabama Poultry Producers, County and Regional Extension Agents, Researchers and the SMREC staff. Farmers in attendance were very receptive to the idea of producing livestock feed and other value added products from materials available on-farm, with little to no additional cost. (5) Presentation was made to Heifer International representatives at the Heifer Ranch near Little Rock, AR in April 2008. Interest was shown in applying our concept in Peru, Ecuador, and Mexico. (6) Presentation was also made to a Food and Agriculture Organization (FAO) representative in Little Rock, AR in April 2008. FAO is interested in the results of the project and has been kept up to date on the project via email and telephone correspondence. (7) The research associate working on the project attended the Fungi Perfecti's mycoremediation workshop. Mycoremediation aspects covered in the workshop included, strain selection, environmental influences on cultivation and remediative efficacy, specific remediative properties of a number of fungi strains, including P. ostreatus. (8) Research Associate also attended the Mycoremediation section of the Sustainable Agriculture Working Group (SAWG) conference. PARTICIPANTS: Jayme Oates, Research Associate, Fisheries and Allied Aquaculture, Auburn University, Auburn, AL. Puneet Srivastava, Assistant Processor, Biosystems Engineering, Auburn University, Auburn, AL. (PD). Kathy S. Lawrence, Associate Professor, Entomology and Plant Pathology, Auburn University, Auburn, AL. (Co-PD). The project provided training to two graduate students. TARGET AUDIENCES: Poultry Producers, Mushroom Growers, Soil and Water Conservation Agencies, State Extension Services. The effort will help better manage broiler litter, reduce threat of soil and water contamination, and generated value added products from poultry litter (e.g., mushroom fruits and mushroom compost). PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
(1) Out of the three strains tested, Pleurotus ostreatus was determined to be the best cultivation choice for Alabama farmers due to its fast and hardy growth habits, and tolerance to extreme environmental conditions (temperature, moisture, light, CO2 levels). (2) All three fungi grew successfully on both woodchip and straw autoclaved substrates at the three levels of poultry manure concentrations (30:1 C:N, 15:1 C:N, and 10:1 C:N). However, growth slowed with increasing nitrogen concentrations, and subsequent mushroom fruiting on the highest nitrogen substrate occurred 1-4 weeks after the substrates containing lower nitrogen concentrations.

Publications

No publications reported this period