The aquatic bird, egret, could carry antibiotic resistance (AR) from a contaminated waterway (Jin River, Chengdu, China) into the surrounding environment (Wangjianglou Park). A systematic study was carried out on the unique environmental dissemination mode of AR mediated by birds. The minimum inhibitory concentrations of various antibiotics against the environmental Escherichia coli isolates were used to evaluate the bacterial AR at the environmental locations where these isolates were recovered, i.e., the Jin River water, the egret feces, the park soil, and the campus soil. The level of AR in the park soil was significantly higher than that in the campus soil that was seldom affected by the egrets, which suggested that the egrets mediated the transportation of AR from the polluted waterway to the park. Genotyping of the resistant E. coli isolates via repetitive-element PCR gave no strong correlation between the genotypes and the AR patterns of the bacteria. So, the transfer of resistant strains should not be the main mode of AR transportation in this process. The results of real-time PCR revealed that the abundance of antibiotic resistance genes (ARGs) and mobile genetic element (MGE) sequences (transposase and integrase genes) declined along the putative transportation route. The transportation of ARGs could be due to their linkage with MGE sequences, and horizontal gene transfer should have contributed to the process. The movable colistin-resistance gene mcr-1 was detected among the colistin-resistant E. coli strains isolated from the river water and the egret feces, which indicated the possibility of the environmental dissemination of this gene. Birds, especially the migratory birds, for the role they played on the dissemination of environmental AR, should be considered when studying the ecology of AR.

• Migratory birds are a major source of antibiotic resistance around Qinghai Lake. • Horizontal transfer of antibiotic resistance genes is confined in migratory birds. • There is high co-selection potential between antibiotic resistance genes. • Migratory birds deserve more attention for spreading antibiotic resistance.

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We sequenced the whole genomes of three mcr-1-positive multidrug-resistant E. coli strains, which were previously isolated from the environment of egret habitat (polluted river) and egret feces. The results exhibit high correlation between antibiotic-resistant phenotype and genotype among the three strains. Most of the mobilized antibiotic resistance genes (ARGs) are distributed on plasmids in the forms of transposons or integrons. Multidrug-resistant (MDR) regions of high homology are detected on plasmids of different E. coli isolates. Therefore, horizontal transfer of resistance genes has facilitated the transmission of antibiotic resistance between the environmental and avian bacteria, and the transfer of ARGs have involved multiple embedded genetic levels (transposons, integrons, plasmids, and bacterial lineages). Inspired by this, systematic metadata analysis was performed for the available sequences of mcr-1-bearing plasmids. Among these plasmids, IncHI2 plasmids carry the most additional ARGs. The composition of these additional ARGs varies according to their geographical distribution. The phylogenetic reconstruction of IncI2 and IncX4 plasmids provides the evidence for their multiregional evolution. Phylogenetic analysis at the level of mobile genetic element (plasmid) provides important epidemiological information for the global dissemination of mcr-1 gene. Highly homologous mcr-1-bearing IncI2 plasmids have been isolated from different regions along the East Asian-Australasian Flyway, suggesting that migratory birds may mediate the intercontinental transportation of ARGs.

Antibiotics released in the environment have attracted great attention. The environmental emission control of antibiotics should be based on the degree of their negative impacts on the environment and ecology. Here, we conducted a series of soil slurry microcosm experiments to investigate the interactions between antibiotics and the soil bacterial community. In the soil slurry, distinctive behaviors were observed for different antibiotics. Betalactams (ampicillin and ceftriaxone) experienced fast biodegradation. Kanamycin was adsorbed on soil particles soon after its addition. Nalidixic acid was stable throughout the experimental period (164 h). The main inactivation mechanism of tetracycline was deduced to be hydrolysis. Bacterial communities in slurries with or without antibiotic-treatment were profiled via high-throughput Illumina sequencing of the 16S rRNA gene. Unstable (ceftriaxone) and adsorbed (kanamycin) antibiotics show minor or negligible influences on the soil bacterial community. Stable antibiotics (nalidixic acid and tetracycline) have significantly affected the structure of the bacterial community. Most of enriched bacterial genera by various antibiotics belong to the same phylum, Proteobacteria. Inhibited bacterial phyla by nalidixic acid are Firmicutes and Bacteroidetes, while those inhibited by tetracycline are Firmicutes, Bacteroidetes and Cyanobacteria. According to the PICRUSt prediction of metagenome, influence of antibiotics on overall metabolic function of the bacterial community is rather limited. This study has provided valuable information, from a phylogenetic viewpoint, about the influence of high concentration of antibiotics on soil bacterial community.