Needles in haystacks: monitoring the potential escape of bioaerosolised antibacterial resistance genes from wastewater treatment plants with air and phyllosphere sampling.

Wastewater treatment plants are well-known point sources of emissions of antibacterial resistance genes (ARGs) into the environment. Although most work to date has focused on ARG dispersal via effluent, aerial dispersal in bioaerosols is a poorly understood, but likely important vector for ARG dispersal. Recent evidence suggests that ARG profiles of the conifer needle phyllosphere could be used to measure bioaerosol dispersal from anthropogenic sources. Here, we assessed airborne dispersal of ARGs from wastewater treatment plants in Wales, UK and Quebec, Canada, using conifer needles as passive bioaerosol monitors. ARG profiles of wastewater were compared to those of conifer phyllosphere using high-throughput qPCR. ARG richness was significantly lower in conifer phyllosphere samples than wastewater samples, though no differences were observed across the dispersal gradients. Mean copy number of ARGs followed a similar trend. ARG profiles showed limited, but consistent patterns with increasing distance from wastewater treatment plants, but these did not align with those of wastewater samples. For example, proportional abundance of aminoglycosides decreased over the dispersal gradient in Wales, whereas mobile genetic elements showed the inverse relationship. In summary, while distinct ARG profiles exist along dispersal gradients, links to those of wastewater were not apparent.

Effect of a probiotic and an antibiotic on the mobilome of the porcine microbiota.

Introduction: To consider the growing health issues caused by antibiotic resistance from a "one health" perspective, the contribution of meat production needs to be addressed. While antibiotic resistance is naturally present in microbial communities, the treatment of farm animals with antibiotics causes an increase in antibiotic resistance genes (ARG) in the gut microbiome. Pigs are among the most prevalent animals in agriculture; therefore, reducing the prevalence of antibiotic-resistant bacteria in the pig gut microbiome could reduce the spread of antibiotic resistance. Probiotics are often studied as a way to modulate the microbiome and are, therefore, an interesting way to potentially decrease antibiotic resistance. Methods: To assess the efficacy of a probiotic to reduce the prevalence of ARGs in the pig microbiome, six pigs received either treatment with antibiotics (tylvalosin), probiotics (Pediococcus acidilactici MA18/5M; Biopower® PA), or a combination of both. Their faeces and ileal digesta were collected and DNA was extracted for whole genome shotgun sequencing. The reads were compared with taxonomy and ARG databases to identify the taxa and resistance genes in the samples. Results: The results showed that the ARG profiles in the faeces of the antibiotic and combination treatments were similar, and both were different from the profiles of the probiotic treatment (p < 0.05). The effects of the treatments were different in the digesta and faeces. Many macrolide resistance genes were detected in a higher proportion in the microbiome of the pigs treated with antibiotics or the combination of probiotics and antibiotics. Resistance-carrying conjugative plasmids and horizontal transfer genes were also amplified in faeces samples for the antibiotic and combined treatments. There was no effect of treatment on the short chain fatty acid content in the digesta or the faeces. Conclusion: There is no positive effect of adding probiotics to an antibiotic treatment when these treatments are administered simultaneously.

Antimicrobial Resistance in the Environment: Towards Elucidating the Roles of Bioaerosols in Transmission and Detection of Antibacterial Resistance Genes.

Antimicrobial resistance (AMR) is continuing to grow across the world. Though often thought of as a mostly public health issue, AMR is also a major agricultural and environmental problem. As such, many researchers refer to it as the preeminent One Health issue. Aerial transport of antimicrobial-resistant bacteria via bioaerosols is still poorly understood. Recent work has highlighted the presence of antibiotic resistance genes in bioaerosols. Emissions of AMR bacteria and genes have been detected from various sources, including wastewater treatment plants, hospitals, and agricultural practices; however, their impacts on the broader environment are poorly understood. Contextualizing the roles of bioaerosols in the dissemination of AMR necessitates a multidisciplinary approach. Environmental factors, industrial and medical practices, as well as ecological principles influence the aerial dissemination of resistant bacteria. This article introduces an ongoing project assessing the presence and fate of AMR in bioaerosols across Canada. Its various sub-studies include the assessment of the emissions of antibiotic resistance genes from many agricultural practices, their long-distance transport, new integrative methods of assessment, and the creation of dissemination models over short and long distances. Results from sub-studies are beginning to be published. Consequently, this paper explains the background behind the development of the various sub-studies and highlight their shared aspects.

Production of composted recycled manure solids from a Canadian dairy farm: Impact on microbial air quality in experimental conditions.

Recycled manure solids (RMS) produced in dairy farms from fresh manure need to be sanitized before using them as bedding material. However, the impact on air quality of composting RMS remains unknown. Four RMS composting methods were tested during a 10-day aging of piles in experimental chambers: static windrow (SW), turned windrow (TW), SW following drum composting for 24 h (DC24) or SW following drum composting for 72 h (DC72). Air samples were collected using a SASS®3100 Dry Air Sampler on days 0 (pilling of the RMS), 5, and 10. Bacteria (16S rRNA genes), Penicillium/Aspergillus, A fumigatus, and 11 human pathogenic bacteria (e.g. Klebsiella pneumonia) were quantified by qPCR while endotoxins and dust particles were, respectively, measured by LAL assays and with a DustTrakTM DRX Aerosol Monitor. On day 0, RMS produced by SW and TW yielded the lowest concentrations of airborne bacteria, while DC24 resulted in the lowest levels of Penicillium/Aspergillus and dust particles. SW method led on day 5 to the lowest concentration of bacteria and Penicillium/Aspergillus, and DC24 and DC72 to the lowest concentration of airborne dust. On day 10, SW and TW piles were associated with the lowest levels of Penicillium/Aspergillus and dust particles. A significant difference was observed between concentration of airborne bacteria, Penicillium/Aspergillus and endotoxins before and during the turnover of TW piles. None of the studied human pathogens was detected in the air samples. Results of the present study suggest that SW and TW are the most promising methods for the production of composted RMS with respect to microbial air quality. However, the experimental chambers do not accurately represent commercial dairy barns and further research on these composting methods is necessary. Finally, the study highlights that bedding material and its management may be determinant factors for air quality in dairy barns.Implications: The research evaluated the impact on microbial air quality of composting recycled manure solids (RMS) produced from fresh cow manure. RMS need to be composted or sanitized before using them as bedding material for animals. The impact on animal health of RMS still needs to be confirmed, while the effect on air quality and the health of dairy farmers is unknown. In the present study, microbial air quality associated with four RMS composting methods was investigated. Data revealed that two methods resulted in lower aerosolization of dust particles, endotoxins, molds, and bacteria.