Skip to main content
ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Sustainable Agricultural Systems Laboratory » Research » Publications at this Location » Publication #388079
Research Project: Soil, Crop, and Manure Biochemistry and Molecular Ecology: Bridging Knowledge Gaps in Microbiome Response to Management and Climate Change

Location: Sustainable Agricultural Systems Laboratory

Title: Two-step multi-residue antibiotic extraction method for comparison of antibiotic concentrations in manure as it moves through a manure treatment system

Author
item YARBERRY, ANDREA - Oak Ridge Institute For Science And Education (ORISE)
item POINDEXTER, CARLTON - University Of Maryland
item Rice, Clifford
item LANSING, STEPHANIE - University Of Maryland

Submitted to: Science of the Total Environment
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/22/2022
Publication Date: 12/1/2022
Citation: Yarberry, A., Poindexter, C., Rice, C., Lansing, S. 2022. Two-step multi-residue antibiotic extraction method for comparison of antibiotic concentrations in manure as it moves through a manure treatment system. Science of the Total Environment. https://doi.org/10.3390/antibiotics11121735.
DOI: https://doi.org/10.3390/antibiotics11121735

Interpretive Summary: Use of antibiotics to thwart animal infections in agricultural settings has led to microbial resistance to antibiotics; a serious threat to human medical treatments. Here we developed a method to extract 10 antibiotics in 4 antibiotic classes (tetracycline, sulfonamides, macrolides, and ß-lactam) from farm animal manure substrates. With this extraction method we can now assess the efficacy of farm livestock manure processing technologies for reduction of antibiotic concentrations and develop protocols to determine decay of antibiotics as they are processed. Characterization of the efficacy of these manure processing systems will inform land managers and policy makers of the potential for risk of antibiotic flow through on-farm and off-farm manure processing.

Technical Abstract: Due to the vastly different physiochemical properties of antibiotics administered to dairy cows, antibiotics will distribute differently into the liquid and solid fractions of manure. The purpose of this study was to develop a method to extract 10 antibiotics in 4 antibiotic classes (tetracycline, sulfonamides, macrolides, and ß-lactam) from manure substrates generated through solid-liquid separation. This study systematically manipulated previously published methods to maximize extraction efficiencies in manure. The developed method was a two-step liquid-solid extraction using 10 mL each of 0.1 M EDTA-McIlviane buffer as the first extractant and methanol (MeOH) as the second. After each of the extracts was added, the solutions were vortexed (10 s.), sonicated (15 min.), rotary mixed (15 min.), then centrifuged at 3,300 x g for 20 minutes. The combined extracts were then diluted with HPLC-grade organic free water and cleaned up with solid phase extraction (SPE) using reverse phase (STRATA) cartridges. Antibiotic quantitation was performed by liquid chromatography tandem mass spectrometry (LC-MS/MS) under positive electrospray ionization mode (ESI+) with multiple-reaction monitoring. Total recoveries across manure substrates were 66-131% for tetracyclines, 56% for sulfonamide, 47-55% for macrolides, and 0-57% for ß-lactams. Antibiotics analysis within manure is pivotal for determining appropriate risk assessment for final waste streams.