[Impact of Tetracycline Antibiotic on the Transcriptional Expression of Tetracycline Resistance Genes in Shigella flexneri].

The tetracycline (TC) antibiotic has been widely found in different environmental matrices. The tetracycline resistant bacterium (TRB) of Shigella flexneri was screened and purified from activated sludge, and was then used to study the impact of TC stress on the gene abundances and expression levels of TC resistance genes (TC-ARGs), including tetC, tetO, and tetX, which were respectively quantified by quantitative PCR and reverse transcriptional PCR. Correlations between the TC concentration and gene abundances of TC-ARGs and their expression levels were discussed. The results showed that TC stress had an inhibiting effect on the growth of Shigella flexneri during the entire culture cycle (24 h) and that the growth rate of the bacterial concentration decreased with increasing TC concentration. However, less impact on the gene abundance of TC-ARGs was found. TC stress could promote the expression of TC-ARGs in Shigella flexneri, and the expression levels of tetC, tetO, and tetX genes first increased and then decreased. The correlation results indicated that no significant correlation was observed between the TC concentration and gene abundances of TC-ARGs and their expression levels. Nevertheless, the gene abundances of tetC and tetO were significantly correlated with their expression levels, thus indicating that they can be used to evaluate and assess expression levels to a certain extent.

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

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.