[Effect of Tetracycline Antibiotic on Abundance and Transcriptional Expression Level of Tetracycline Resistance Genes in Activated Sludge]

Abstract

Tetracycline resistant bacteria (TRB) screened from activated sludge were used to study the effect of tetracycline (TC) antibiotic on the transcriptional expression of tetracycline resistance genes (TC-ARGs). The gene abundances of seven TC-ARGs including tetA, tetC, tetG, tetM, tetO, tetW, and tetX, as well as their expression levels, were quantified by quantitative PCR (qPCR) and reverse transcriptional PCR (RT-PCR). The correlations between TC concentrations and gene abundance of TC-ARGs and their expression levels were discussed. The results showed that the gene abundances of tetA, tetG, and tetW generally increased with increasing TC exposure concentrations during the entire culture cycle, whereas other TC-ARGs fluctuated greatly. The impact of TC stress on the transcriptional expression level of different TC-ARGs varied to a great extent. The gene expression of tetA was relatively stable and exhibited an upregulated trend with increasing TC concentrations. When the TC concentration was 100 mg·L^-1, the upregulation of tetA expression was as high as 5.3-fold compared with the control. Under short-term TC stress (one day), the transcriptional expression level was upregulated with increasing TC concentration. The correlation results showed that gene abundances of tetA and tetW correlated significantly with their respective expression levels, indicating that they can evaluate expression levels to a certain extent, which can further mirror functional activities and environmental risks.

Keywords: activated sludge; gene abundance; tetracycline antibiotic; tetracycline resistance gene; transcriptional expression level.