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By performing a microcosm experiment mimicking fertilization, we assessed the dynamic distribution of tetracycline-resistant bacteria (TRB) and corresponding tetracycline resistance genes (TRGs) from pig manure (PM) to the fertilized soil, by culture-dependent methods and PCR detection. Cultivable TRB were most abundant in PM, followed by fertilized soil and unfertilized soil. By restriction fragment length polymorphism (RFLP) analysis, TRB were assigned to 29, 20, and 153 operational taxonomic units (OTUs) in PM, unfertilized soil, and fertilized soil, respectively. After identification, they were further grouped into 19, 12, and 62 species, showing an enhanced diversity of cultivable TRB in the soil following PM application. The proportions of potentially pathogenic TRB in fertilized soil decreased by 69.35% and 41.92% compared with PM and unfertilized soil.  Bacillus cereus  was likely widely distributed TRB under various environments, and  Rhodococcus erythropolis  and  Acinetobacter  sp. probably spread from PM to the soil via fertilization. Meanwhile,  tet L was the most common efflux pump gene in both unfertilized and fertilized soils relative to PM;  tet B(P) and  tet 36 were common in PM, whereas  tet O was predominant in unfertilized and fertilized soil samples. Sequencing indicated that over 65% of randomly selected TRB in fertilized soil with acquired resistance derived from PM.

High diversity and abundance of cultivable tetracycline-resistant bacteria in soil following pig manure application