Phage-Based Biocontrol of Antibiotic-Resistant Bacterium Isolated from Livestock Wastewater Treatment Plant

Abstract

The presence of antibiotic-resistant bacteria (ARB) in receiving water can severely threaten the aquatic environment and human health. The treated effluent containing ARB in some livestock wastewater treatment plants (WWTPs) is returned to the municipal WWTP to reduce the residual ammonia and phosphorus concentrations. ARBs are widespread through wastewater treatment processes and are discharged into river and lake. This study highlights that the isolated lytic phage could reduce ARB isolated from livestock WWTPs and apply phage-based biocontrol in mixed cultures. ARB and lytic phages were isolated from livestock wastewater and used in a batch reactor with diverse cultures. The isolated bacterium was from the Aeromonas species and was resistant to various antibiotics (penicillin, tetracycline, colistin, and kanamycin), indicating multi-drug resistance and biofilm formation. The isolated lytic phage successfully infected Aeromonas species in pure culture and was relatively stable in terms of pH, temperature, and toxic chemicals. The multiplicity of infection (MOI) was examined to determine the proper phage number to kill the host bacterium. The optimal number to control the isolated ARB was a 1:100 phage-to-host ratio. Scanning electron microscopy showed that lytic phages reduced bacterial growth and biofilm formation. Phage-mediated biocontrol was applied in a batch reactor with mixed cultures. Pyrosequencing data from the batch reactor indicated that lytic phages reduced the proportion of the isolated ARB from 65.7 to 20% in 24 h. This study provides evidence for the possible application of lytic phages to control ARB in treated wastewater and an alternative method to prevent the widespread exposure of ARB without producing chemical byproducts.