Many freshwater cyanobacteria, including Microcystis aeruginosa, lack several known antibiotic resistance genes; however,
both axenic and xenic M. aeruginosa strains exhibited high antibiotic resistance against many antibiotics under our tested
concentrations, including colistin, trimethoprim, and kanamycin. Interestingly, axenic PCC7806, although not the xenic
NIBR18 and NIBR452 strains, displayed susceptibility to ampicillin and amoxicillin, indicating that the associated bacteria
in the phycosphere could confer such antibiotic resistance to xenic strains. Fluorescence and scanning electron microscopic
observations revealed their tight association, leading to possible community-level β-lactamase activity. Combinatory treatment
of ampicillin with a β-lactamase inhibitor, sulbactam, abolished the ampicillin resistance in the xenic stains. The
nitrocefin-based assay confirmed the presence of significant community-level β-lactamase activity. Our tested low ampicillin
concentration and high β-lactamase activity could potentially balance the competitive advantage of these dominant species
and provide opportunities for the less competitive species, thereby resulting in higher bacterial diversity under ampicillin
treatment conditions. Non-PCR-based metagenome data from xenic NIBR18 cultures revealed the dominance of blaOXArelated
antibiotic resistance genes followed by other class A β-lactamase genes (AST-1 and FAR-1). Alleviation of ampicillin
toxicity could be observed only in axenic PCC7806, which had been cocultured with β-lactamase from other freshwater
bacteria. Our study suggested M. aeruginosa develops resistance to old-class β-lactam antibiotics through altruism, where
associated bacteria protect axenic M. aeruginosa cells.
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