Co-occurrence of antimicrobials and metals as potential drivers of antimicrobial resistance in swine farms

The application of animal manures to cropland is an important nutrient recycling strategy in many parts of the world. Commonly, aggregated manure wastes contain chemical stressors including veterinary antimicrobials, heavy metals, and antimicrobial resistance genes (ARGs) that can stimulate the development and proliferation of antimicrobial resistance (AMR). While the presence of antimicrobials in manure is well-documented, the co-occurrence of other potentially impactful chemical stressors in swine manure remains underreported. This study quantifies and analyzes correlations between antimicrobials, metals, and certain ARGs present in manure samples from swine farms in Iowa, United States. Relationships between chemical stressors and different stages of swine production or feed composition are also investigated. Results revealed substantial levels of tetracyclines [up to 1,260 µg g−1 dry weight (d.w.) of manure for oxytetracycline] detected in all samples. Tiamulin, two ionophores (monensin and lasalocid), and one macrolide (tilmicosin) were detected at maximum class concentrations of 9.4, 0.547, and 0.472 µg g−1 d.w., respectively. The median relative abundances of ermB and tetM were 0.13 and 0.17 copies g−1 wet weight (w.w.) manure (normalized to 16S gene), respectively. Additionally, high levels of copper (Cu), iron (Fe), and zinc (Zn) were detected in all samples, with maximum concentrations of 887, 1,900, and 2,100 µg g−1 d.w., respectively. Notably, uranium (U) was detected in 11 samples, at concentrations up to 0.77 µg g−1. A global analysis of AMR-stressor relationships using Spearman’s rank correlation indicates Cu, and Ba are the most positively and significantly correlated with cytotoxic anhydrotetracycline (ATC) and/or anhydrochlortetracycline (ACTC) concentrations in all tested facilities (Cu-ATC: ρ = 0.67, p = 0.0093; Cu-ACTC: ρ = 0.75, p = 0.0022; Ba-ATC: ρ = 0.84, p = 0.0002). Interestingly, ermB and tetM genes were strongly, positively correlated to each other (ρ = 0.92, p < 0.0001), suggesting possible co-selection, despite the absence of correlation between ARGs and tetracycline concentrations. This study demonstrates the complexity of interactions between antimicrobials, metals, and ARGs in multiple manure storage pits prior to cropland application.This article is published as Congilosi, Jena L., Joshua S. Wallace, Timothy P. Neher, Adina Howe, Michelle L. Soupir, and Diana S. Aga. "Co-occurrence of antimicrobials and metals as potential drivers of antimicrobial resistance in swine farms." Frontiers in Environmental Science 10 (2022): 1018739. doi: https://doi.org/10.3389/fenvs.2022.1018739. © 2022 Congilosi, Wallace, Neher, Howe, Soupir and Aga. This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/)

Microbial Communities as a Pathway to Improved Woodchip and Corncob Bioreactor Design and Performance

Antibiotics are administered to livestock in animal feeding operations (AFOs) for the control, prevention, and treatment of disease. Manure from antibiotic treated livestock contains unmetabolized antibiotics that provide selective pressure on bacteria, facilitating the expression of anti-microbial resistance (AMR). Manure application on row crops is an agronomic practice used by growers to meet crop nutrient needs; however, it can be a source of AMR to the soil and water environment. This study in central Iowa aims to directly compare AMR indicators in outlet runoff from two adjacent (221 to 229 ha) manured and non-manured catchments (manure comparison), and among three catchments (600 to 804 ha) with manure influence, no known manure application (control), and urban influences (mixed land use comparison). Monitored AMR indicators included antibiotic resistance genes (ARGs) ermB, ermF (macrolide), tetA, tetM, tetO, tetW (tetracycline), sul1, sul2 (sulfonamide), aadA2 (aminoglycoside), vgaA, and vgaB (pleuromutilin), and tylosin and tetracycline resistant enterococci bacteria. Results of the manure comparison showed significantly higher (p<0.05) tetracycline and tylosin resistant bacteria from the catchment with manure application in 2017, but no differences in 2018, possibly due to changes in antibiotic use resulting from the Veterinary Feed Directive. Moreover, the ARG analysis indicated a larger diversity of ARGs at the manure amended catchment. The mixed land use comparison showed the manure amended catchment had significantly higher (p<0.05) tetracycline resistant bacteria in 2017 and significantly higher tylosin resistant bacteria in 2017 and 2018 than the urban influenced catchment. The urban influenced catchment had significantly higher ermB concentrations in both sampling years, however the manure applied catchment runoff consisted of higher relative abundance of total ARGs. Additionally, both catchments showed higher AMR indicators compared to the control catchment. This study identifies four ARGs that might be specific to AMR as a result of agricultural sources (tetM, tetW, sul1, sul2) and optimal for use in watershed scale monitoring studies for tracking resistance in the environment.This article is published as Neher, Timothy P., Lanying Ma, Thomas B. Moorman, Adina C. Howe, and Michelle L. Soupir. "Catchment-scale export of antibiotic resistance genes and bacteria from an agricultural watershed in central Iowa." PLoS One 15, no. 1 (2020): e0227136. doi: https://doi.org/10.1371/journal.pone.0227136. Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted

Catchment-scale export of antibiotic resistance genes and bacteria from an agricultural watershed in central Iowa.

Antibiotics are administered to livestock in animal feeding operations (AFOs) for the control, prevention, and treatment of disease. Manure from antibiotic treated livestock contains unmetabolized antibiotics that provide selective pressure on bacteria, facilitating the expression of anti-microbial resistance (AMR). Manure application on row crops is an agronomic practice used by growers to meet crop nutrient needs; however, it can be a source of AMR to the soil and water environment. This study in central Iowa aims to directly compare AMR indicators in outlet runoff from two adjacent (221 to 229 ha) manured and non-manured catchments (manure comparison), and among three catchments (600 to 804 ha) with manure influence, no known manure application (control), and urban influences (mixed land use comparison). Monitored AMR indicators included antibiotic resistance genes (ARGs) ermB, ermF (macrolide), tetA, tetM, tetO, tetW (tetracycline), sul1, sul2 (sulfonamide), aadA2 (aminoglycoside), vgaA, and vgaB (pleuromutilin), and tylosin and tetracycline resistant enterococci bacteria. Results of the manure comparison showed significantly higher (p<0.05) tetracycline and tylosin resistant bacteria from the catchment with manure application in 2017, but no differences in 2018, possibly due to changes in antibiotic use resulting from the Veterinary Feed Directive. Moreover, the ARG analysis indicated a larger diversity of ARGs at the manure amended catchment. The mixed land use comparison showed the manure amended catchment had significantly higher (p<0.05) tetracycline resistant bacteria in 2017 and significantly higher tylosin resistant bacteria in 2017 and 2018 than the urban influenced catchment. The urban influenced catchment had significantly higher ermB concentrations in both sampling years, however the manure applied catchment runoff consisted of higher relative abundance of total ARGs. Additionally, both catchments showed higher AMR indicators compared to the control catchment. This study identifies four ARGs that might be specific to AMR as a result of agricultural sources (tetM, tetW, sul1, sul2) and optimal for use in watershed scale monitoring studies for tracking resistance in the environment

Finding What Is Inaccessible: Antimicrobial Resistance Language Use among the One Health Domains

The success of a One Health approach to combating antimicrobial resistance (AMR) requires effective data sharing across the three One Health domains (human, animal, and environment). To investigate if there are differences in language use across the One Health domains, we examined the peer-reviewed literature using a combination of text data mining and natural language processing techniques on 20,000 open-access articles related to AMR and One Health. Evaluating AMR key term frequency from the European PubMed Collection published between 1990 and 2019 showed distinct AMR language usage within each domain and incongruent language usage across domains, with significant differences in key term usage frequencies when articles were grouped by the One Health sub-specialties (2-way ANOVA; p &lt; 0.001). Over the 29-year period, “antibiotic resistance” and “AR” were used 18 times more than “antimicrobial resistance” and “AMR”. The discord of language use across One Health potentially weakens the effectiveness of interdisciplinary research by creating accessibility issues for researchers using search engines. This research was the first to quantify this disparate language use within One Health, which inhibits collaboration and crosstalk between domains. We suggest the following for authors publishing AMR-related research within the One Health context: (1) increase title/abstract searchability by including both antimicrobial and antibiotic resistance related search terms; (2) include “One Health” in the title/abstract; and (3) prioritize open-access publication

Quantifying SARS-CoV-2 spread in Switzerland based on genomic sequencing data

Pathogen genomes provide insights into their evolution and epidemic spread. We sequenced 1,439 SARS-CoV-2 genomes from Switzerland, representing 3-7% of all confirmed cases per week. Using these data, we demonstrate that no one lineage became dominant, pointing against evolution towards general lower virulence. On an epidemiological level, we report no evidence of cryptic transmission before the first confirmed case. We find many early viral introductions from Germany, France, and Italy and many recent introductions from Germany and France. Over the summer, we quantify the number of non-traceable infections stemming from introductions, quantify the effective reproductive number, and estimate the degree of undersampling. Our framework can be applied to quantify evolution and epidemiology in other locations or for other pathogens based on genomic data

Swiss public health measures associated with reduced SARS-CoV-2 transmission using genome data.

Genome sequences from evolving infectious pathogens allow quantification of case introductions and local transmission dynamics. We sequenced 11,357 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genomes from Switzerland in 2020 - the sixth largest effort globally. Using a representative subset of these data, we estimated viral introductions to Switzerland and their persistence over the course of 2020. We contrasted these estimates with simple null models representing the absence of certain public health measures. We show that Switzerland's border closures de-coupled case introductions from incidence in neighboring countries. Under a simple model, we estimate an 86-98% reduction in introductions during Switzerland's strictest border closures. Furthermore, the Swiss 2020 partial lockdown roughly halved the time for sampled introductions to die out. Last, we quantified local transmission dynamics once introductions into Switzerland occurred, using a phylodynamic model. We found that transmission slowed 35-63% upon outbreak detection in summer 2020, but not in fall. This finding may indicate successful contact tracing over summer before overburdening in fall. The study highlights the added value of genome sequencing data for understanding transmission dynamics