Effect of fertilizer types on antibiotic resistance genes and bacterial community in vegetable fields

Farmland ecosystems are essential sources and sinks of antibiotic resistance genes (ARGs), and the application of livestock manure is a major contributor to ARGs in soil. The massive application of livestock manure to vegetable fields has intensified the pollution caused by ARGs in soil. Raw consumption of edible vegetables is one of the most direct ways to introduce ARGs from the soil–plant system to humans, which poses a potential threat to human health. However, few studies have investigated the effects of different fertilizer types on ARGs and bacterial communities in vegetable fields. In this study, 21 soil samples (0–20 cm) were collected from vegetable fields in Hebei Province using different fertilizer types (fresh fowl manure, fresh sheep manure, fresh cattle manure, commercial organic fertilizer, and chemical fertilizer). The distributions and characteristics of ARGs and bacterial communities in vegetable fields were investigated using real-time quantitative polymerase chain reaction (PCR) and high-throughput sequencing techniques. Eight tetracycline resistance genes (tetA, tetC, tetG, tetL, tetO, tetM, tetW, and tetQ), two sulfonamide resistance genes (sul1 and sul2), and one intI1 gene were detected in all vegetable fields. The absolute abundance of sulfonamide resistance genes (9.96×10⁹ copies·g⁻¹ in dry soil) was significantly higher than that of tetracycline resistance genes (1.07×10⁹ copies·g⁻¹ in dry soil). The application of livestock manure and chemical fertilizer both significantly increased the abundance of ARGs in vegetable fields. The highest abundance of ARGs (6.34×10⁹ copies∙g⁻¹ in dry soil) was found in vegetable fields with higher chemical fertilizer amendment, while the lowest abundance of ARGs (3.09×10⁸ copies∙g⁻¹ in dry soil) was found in vegetable soil with commercial organic fertilizer. In addition, the Shannon and Chao1 indices, representing the α diversity of the soil bacterial community, were significantly higher in soil fertilized with livestock manure compared to high-chemical fertilizer application but not in low-chemical fertilization soil, indicating that livestock manure application significantly increased the abundance and diversity of the soil bacterial community. Pearson’s correlation analysis showed that soil bacterial community structure was an important factor influencing the distribution of ARGs. Proteobacteriota, Bacteroidota, Actinobacteriota, and Firmicutes were the dominant potential hosts of ARGs and were significantly correlated with sulfonamide and tetracycline resistance genes (P<0.05). The distribution of ARGs was also affected by soil organic matter and total nitrogen content. The intI1 gene had significant and positive correlations with the sul2, tetG, tetQ, and tetW genes, suggesting its crucial role in ARGs dissemination. In the present study, the use of higher concentrations of chemical fertilizers led to a significantly increased abundance of ARGs in the soil of vegetable fields, whereas the application of commercial organic fertilizers had the least effect on ARGs abundance. This study serves as a guide for evaluating the status of ARGs pollution in vegetable fields with different fertilizer types.