Source: CORNELL UNIVERSITY submitted to
AGROCHEMICAL IMPACTS ON HUMAN AND ENVIRONMENTAL HEALTH: MECHANISMS AND MITIGATION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1021148
Grant No.
(N/A)
Project No.
NYC-187801
Proposal No.
(N/A)
Multistate No.
W-3045
Program Code
(N/A)
Project Start Date
Oct 1, 2019
Project End Date
Sep 30, 2020
Grant Year
(N/A)
Project Director
Hay, AN.
Recipient Organization
CORNELL UNIVERSITY
(N/A)
ITHACA,NY 14853
Performing Department
Microbiology
Non Technical Summary
Though recently banned, triclosan is one of the most abundant personal care products in land-applied biosolids. Stenotrophomonas bacteria are enriched in soil receiving triclosan, but little is known about their impact on triclosan's fate. We will screen Stenotrophomonas from three agricultural soils receiving biosolids from different parts of the country for triclosan resistance and degradation.
Animal Health Component
0%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71101101100100%
Knowledge Area
711 - Ensure Food Products Free of Harmful Chemicals, Including Residues from Agricultural and Other Sources;

Subject Of Investigation
0110 - Soil;

Field Of Science
1100 - Bacteriology;
Goals / Objectives
Characterize abiotic and biotic reaction mechanisms, transformation rates, and fate in agricultural and natural ecosystems. Determine adverse impacts from agrochemical exposure to cells, organisms, and ecosystems.
Project Methods
1) Evaluate triclosan resistance and degradation: Triclosan resistance will be determined using the minimum inhibitor concentration (MIC) microdilution method (doi.org/10.1093/jac/dkp271). Analysis of triclosan degradation by pure cultures of Stenotrophomonas will be performed as described previously for our work with Sphingomonas using GC/MS of ethylacetate extracts of culture supernatant exposed to triclosan concentrations that are ½ or less of the MIC (doi.org/10.1007/s10532-015-9730-9).2) Enumerate Stenotrophomonas in agricultural soils and biosolids: Stenotrophomonas will be enumerated and isolated from soils and biosolids using VIA selective medium (doi.org/10.1111/j.1365-2672.2011.05120.x)3) Quantify the concentration of triclosan in the biosolids obtained biannually from each of the regions. The concentration of triclosan in biosolids will be determine by GC/MS as we previously described (doi.org/10.1016/j.scitotenv.2012.06.097).4) Perform genome sequencing and comparative genomics of triclosan resistant and/or degrading Stenotrophomonas isolates obtained from soils from the four regions: Genome sequencing and analysis will be performed on DNA extracted from axenic cultures obtain in objective 2 and will be conducted as described in our earlier work describing the sequencing and analysis of Sporosarcina (DOI: 10.1128/genomeA.00096-16). Comparative physiology will be done by screening genomic libraries of Stenotrophomonas DNA in E. coli as well as transposon and site-directed mutagenesis of Stenotrophomonas isolates to identify genetic determinants required for triclosan resistance and degradation (doi.org/10.1007/s10532-015-9730-9).

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

Outputs
Target Audience:The target audience was researchers and extension agents. Changes/Problems:The project was interrupted when only 5 months along by the onset of the SARS-CoV2 pandemic. All lab work ceased so the undergrad student and technician were mentored in bioinformatic analyses that laid the ground work for a resubmitted proposal (December 2020). The pending proposalwas broadened beyond triclosan and proposed measuring the impact of 3 widelyapplied herbicides on enrichment of antibiotic resistant Stenotrophomonas from agricultural soils. What opportunities for training and professional development has the project provided?The project provided training for one undergraduate who learned techniques in soil microbiology, molecular biology and bioinformatics. A technician also received professional development training in the use of bioinformatics and in project management. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We're unable to make progress on Goal1 during to COVID related lab closures. For Goal 2 we were able to determine that triclosan added to soil increased the number of Stenotrophomonas that are resistant to multiple antibiotics. This suggests that triclosan additions to soil do have the potential to cause adverse impacts by increasing the size of the antibiotic resistance pool.

Publications