Biosolids Amendment Enriches Soil Resistome, Virulence Factors, and ESKAPE Pathogens: A Longitudinal Metagenomic Study

The spread of antibiotic resistance poses a significant challenge to public health worldwide. Wastewater treatment utilities are reservoirs of antibiotic-resistant bacteria and antibiotic resistance genes (ARGs). This study investigates the impact of biosolids amendment on the soil microbiome, resistome, virulence factors, and ESKAPE pathogens during carrot cultivation in a replicated greenhouse study. Metagenomic sequencing revealed that biosolids amendment increased the richness of microbial genera, ARGs, and virulence factors in soil. The relative abundance of ESKAPE pathogens, including Enterococcus faecium , Staphylococcus aureus , Klebsiella pneumoniae , Acinetobacter baumanii , Pseudomonas aeruginosa , and Enterobacter spp., was notably higher in biosolids-amended soils. These pathogens persisted throughout the 11-week cultivation period, raising concerns about the potential for horizontal gene transfer and the spread of antibiotic resistance. The study also identified significant co-occurrences between microbial genera and ARGs, which can suggest the possibility of the microbial taxa harboring the co-occurring ARGs. These findings highlight the importance of continued research and monitoring to ensure the safe and effective use of biosolids in agriculture.