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ARS Home » Plains Area » Lincoln, Nebraska » Agroecosystem Management Research » Research » Publications at this Location » Publication #421706
Research Project: Managing Manure as a Soil Resource for Improved Biosecurity, Nutrient Availability, and Soil Sustainability

Location: Agroecosystem Management Research

Title: Presence of antimicrobial resistance genes in organomineral fertilizer

Author
item TEIXEIRA, GLAUCO MARIANO, - University Of North Alabama
item Condon, Justine
item Durso, Lisa
item BECRAFT, ERIC - University Of North Alabama

Submitted to: Protocols.io
Publication Type: Research Technical Update
Publication Acceptance Date: 12/13/2024
Publication Date: 12/13/2024
Citation: Teixeira, G., Condon, J.C., Durso, L.M., Becraft, E.D. 2024. Presence of antimicrobial resistance genes in organomineral fertilizer. Protocols.io. 1. https://doi.org/10.17504/protocols.io.eq2ly67eegx9/v1.
DOI: https://doi.org/10.17504/protocols.io.eq2ly67eegx9/v1

Interpretive Summary: Antibiotic resistance threatens the continued utility of antibiotic drugs for treating infectious diseases caused by bacteria. Some types of infectious disease causing bacteria are found naturally in feces from humans and animals, and there is concern that when these bacteria carry the DNA instructions for antibiotic resistance (antibiotic resistance genes), they can prolong the length of an illness, or result in death where once the patient might have survived. However, human feces (in the form of biosolids) and animal manures are a rich source of nutrients for the soil that have been used as fertilizer since prehistoric times. They have drawbacks, such as being hard to transport, and being lacking in some nutrients needed for optimal crop yields. Today mineral and synthetic fertilizers are more common in large scale agriculture, but there is growing interest in a novel new fertilizer that combines the advantages of both the organic (biosolids and manures) and mineral fertilizers. This protocol describes assays used to test these new fertilizer formulations for the presence of specific suite of antibiotic resistance genes.

Technical Abstract: This protocol details the steps for detecting various antibiotic resistance genes (ARGs) in organomineral fertilizers, including tetA, tetX, tetO, ermB, sul1, blaCTX-M-32, and intI1, using PCR and gel electrophoresis. Starting with DNA extraction from soil samples, the method describes PCR reaction preparation with specific primers and controls, followed by visualization of results on an agarose gel. Ethidium bromide and SYBR Safe are used for staining, with appropriate safety precautions. This technique allows for the identification of target resistance genes, indicated by the presence of corresponding DNA bands under UV analysis.