Fate of potential indicator antimicrobial resistance genes (ARGs) and bacterial community diversity in simulated manure-soil microcosms.

The aim of this study was to investigate the fate of nine potential indicator antimicrobial resistance genes (ARGs) (sul1, sul2, tetB, tetM, ermB, ermF, fexA, cfr, intI1) and the diversity of bacterial communities in response to poultry manure applications to arable soil over a 90 day period. Quantitative real time PCR and Illumina high-throughput sequencing of 16S rDNA gene were used to quantify and trace ARG fate. The levels of all genes dramatically decreased over time and intI1, sul1, sul2 and tetM always had the greatest abundance and lowest dissipation rates. This indicated that more effort should be focused on the ARG elimination from manure rather than waiting for subsequent attenuation in the environment. Our sequencing results documented dramatic changes in the microbial community structure and diversity during these experiments. In poultry manure groups, Bacteroidetes and Actinobacteria were the two dominant phyla while Acidobacteria dominated the control groups. Moreover, the relative abundance of genera Corynebacterium, Pseudomonas, Ochrobactrum, Actinomadura and Bacillus, which contained potential opportunistic pathogens, changed over time suggesting that poultry manure not only strongly influenced bacterial community composition, but also selected specific bacterial communities. This study provides a glimpse of ARG fates and bacterial community diversity in soil after the application of poultry manure.

Dissemination of resistance genes in duck/fish polyculture ponds in Guangdong Province: correlations between Cu and Zn and antibiotic resistance genes.

Duck/fish polyculture farming is a typical farming model in the Pearl River delta in southern China. We examined soil, water, and sediment samples from three duck-fish farms in Guangdong Province in September and December 2014. We determined the abundance of three metal resistance genes, 16S rDNA, and 23 antibiotic resistance genes encoding resistance to tetracycline, sulfonamides, quinolones, chloramphenicol, and ß-lactamases. Microbial community structure was quantified by Illumina high-throughput sequencing of 16S rDNA genes. We found a prevalence of antibiotic resistance genes and the sul1, sul2, tetA, tetM, aac(6')-Ib, and floR genes were the most abundant. Levels of Cu and Zn were significantly correlated with numerous ARG types and sul2, floR, and tetM were identified as potential antibiotic resistance gene indicators. Cu levels were significantly and positively correlated with the relative abundance of sul3, tetT, tetW, qnrB, qnrS, fexB, sul1, sul2, tetM, and qnrA. Zn was significantly correlated to relative abundance of sul2, sul3, tetM, tetA, tetT, tetW, qnrA, qnrB, qnrS, aac(6')-Ib, qepA, blaSHV, cmlA, floR, fexA, cfr, and fexB. The levels of Acinetobacter, Brevibacillus, and Wautersiella showed significant positive correlations with metal resistance genes as well as qnrB, oqxA, oqxB, and blaSHV (p > 0.8). Sphingobacterium, Flavobacterium, Acidothermus, and Corynebacterium had significant correlations with abundance of tetracycline resistance genes, sulfonamide resistance genes, blaTEM, blaCTX, and cfr (p > 0.8). Sphingobacterium, Flavobacterium, Acidothermus, and Corynebacterium were most abundant in soil samples while Acinetobacter, Brevibacillus, and Wautersiella were most abundant in water samples. Dissemination of antibiotic resistance genes in aquaculture environments is extensive and tracing their origins is necessary to establish risk assessment methods required for aquatic environmental protection.