Factors influencing the fate of antibiotic resistance genes during thermochemical pretreatment and anaerobic digestion of pharmaceutical waste sludge

Highlights: • Factors impact on ARGs during pharmaceutical sludge treatment were studied. • Multiple linear regression was used to assess the importance of factors on ARGs. • Alkalinity showed more influence on ARGs than other sludge features. • Bacterial diversity rather than community composition affected ARGs. • Interaction effect and contributions of each factor on ARGs were elucidated. The prevalence of antibiotic resistance genes (ARGs) in waste sludge, especially for the pharmaceutical waste sludge, presents great potential risks to human health. Although ARGs and factors affecting their spreading are of major importance for human health, the factors influencing the fate of ARGs during sludge treatment, especially for pharmaceutical sludge treatment are not yet well understood. In order to be able to minimize ARGs spreading, it is important to find what is influencing their spreading. Therefore, certain factors, such as the sludge characteristics, bacterial diversity and community composition, and mobile genetic elements (MGEs) during the advanced AD of pharmaceutical sludge with different pretreatments were studied, and their affinity with ARGs was elucidated by Spearman correlation analysis. Furthermore, multiple linear regression was introduced to evaluate the importance of the various factors. Results showed that 59.7%–88.3% of the variations in individual ARGs and total ARGs can be explained by the corresponding factors. Bacterial diversity rather than specific bacterial community composition affected the fate of ARGs, whereas alkalinity was the most important factor on ARGs among all sludge characteristics investigated in this study. Besides, 66.4% of variation of total ARGs was driven by the changes of MGEs. Multiple linear regression models also reveal the collective effect of these factors on ARGs, and the contributions of each factor impact on ARGs. This study provides more comprehension about the factors impact on the fate of ARGs during pharmaceutical sludge treatment, and offers an approach to evaluate the importance of each factor, which method could be introduced for evaluation of factors influencing ARGs during other types of sludge or wastewater treatment.