Antimicrobial resistance in plant endophytes associated with poultry-manure application revealed by selective culture and whole genome sequencing

Abstract

Poultry manure is widely used as organic fertilizer in agriculture during the cultivation of crops, but the persistent high-level use of antibiotics in poultry production has raised concerns about the selection for reservoirs of antimicrobial resistance genes (ARGs). Previous studies have shown that the addition of poultry manure can increase the abundance of genes associated with resistance to tetracyclines, aminoglycosides, fluoroquinolones, sulfonamides, bacitracin, chloramphenicol, and macrolide-lincosamide-streptogramin in soil and plants. Understanding the microbial populations that harbor these ARGs is important to identify microorganisms that could enter the human food chain. Here, we test the hypothesis that environmental exposure to poultry manure increases the occurrence of antimicrobial resistance (AMR) in plant endophytes using selective culture, phenotypic Antibiotic Susceptibility Testing (AST), phylogenetic analysis, and whole genome sequencing (WGS). Endophytes from poultry manure treated Sorghum bicolor (L.) Moench plant root and stem samples showed increased phenotypic and genotypic resistance against multiple antibiotics compared to untreated controls. Comparison of AMR phenotype-to-genotype relationships highlighted the detection of multi-drug resistant (MDR) plant endophytes, demonstrating the value of genomic surveillance for emerging drug-resistant pathogens. The increased occurrence of ARGs in poultry manure-exposed endophytes highlights the need for responsible antibiotic use in poultry and animal farming to reduce contamination of ecological niches and transgression into endophytic plant microbiome compartments. It also emphasizes the requirement for proper manure management practices and vigilance in monitoring and surveillance efforts to tackle the growing problem of antibiotic resistance and preserve the efficacy of antibiotics for human and veterinary medicine.