The ability of multidrug-resistant Escherichia coli to adapt and grow in a wide range of different environmental conditions may be crucial to the global spread of antimicrobial resistance. The aim of this study was to evaluate the survival ability of 54 multidrug-resistant E. coli strains, isolated from three different biotopes (clinical setting, gull intestine, river water) when subjected to variations in pH (from 3 to 11) and salinity (from 0.5% to 6% of NaCl) and to nutrient deprivation. The growth of each isolate as well as of a reference strain was assessed during 168 h in every varying condition. Slight variations in the growth ability under some abiotic stress factors were recorded among the isolates from the different biotopes. Multidrug-resistant isolates from gull feces were found to be the more tolerant to environmental abiotic changes, while isolates from river water were the less tolerant. In addition, it was notorious that the carriage of antimicrobial resistance has a clear fitness cost in comparison with the susceptible (reference) strain, highlighting the necessity of reducing the selective pressure exerted by antibiotics. This study underlines the ecological hardness of multidrug-resistant E. coli isolates with a consequent ability to reach and colonize new host and environments.