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Gentamicin (Gm) is an aminoglycoside commonly used to treat bacterial infections such as tularemia – the disease caused by  Francisella tularensis . In addition to being pathogenic,  F. tularensis  is found in environmental niches such as soil where this bacterium likely encounters Gm producers ( Micromonospora  sp.). Here we show that  F. tularensis  exhibits increased resistance to Gm at ambient temperature (26°C) compared to mammalian body temperature (37°C). To evaluate whether  F. tularensis  was less permeable to Gm at 26°C, a fluorescent marker [Texas Red (Tr)] was conjugated with Gm, yielding Tr-Gm. Bacteria incubated at 26°C showed reduced fluorescence compared to those at 37°C when exposed to Tr-Gm suggesting that uptake of Gm was reduced at 26°C. Unconjugated Gm competitively inhibited uptake of Tr-Gm, demonstrating that this fluorescent compound was taken up similarly to unconjugated Gm. Lysates of  F. tularensis  bacteria incubated with Gm at 37°C inhibited the growth of  Escherichia coli  significantly more than lysates from bacteria incubated at 26°C, further indicating reduced uptake at this lower temperature. Other facultative pathogens ( Listeria monocytogenes  and  Klebsiella pneumoniae ) exhibited increased resistance to Gm at 26°C suggesting that the results generated using  F. tularensis  may be generalizable to diverse bacteria. Regulation of the uptake of antibiotics provides a mechanism by which facultative pathogens survive alongside antibiotic-producing microbes in nature.

Temperature-Dependent Gentamicin Resistance of Francisella tularensis is Mediated by Uptake Modulation