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<p>Abstract</p> <p>Background</p> <p>Frequent and indiscriminate use of antibiotics has led to the development of multi-drug resistant bacterial strains. It necessitates the exploitation of alternative therapeutic strategies. In order to reduce the dose of antibiotic required and to decrease the associated side effects, the present study was aimed at evaluating the synergism, if any, between a conventional antibiotic, co-trimoxazole (CTZ)) and cell free supernatant (CFS) of a probiotic (<it>L. plantarum) </it>against <it>S. </it>Typhimurium NCTC 74. This antimicrobial combination was selected on the basis of antibiotic susceptibility pattern of <it>Salmonella </it>and <it>L. plantarum</it>.</p> <p>Methods</p> <p>The synergy was evaluated in terms of size of zone of inhibition, fractional inhibitory concentration index, time-kill assay (<it>in-vitro) </it>as well as macrophage functions (<it>ex-vivo</it>).</p> <p>Results</p> <p>The concentration producing the same or higher antibacterial effect (size of zone of inhibition) was reduced to half when both the agents were used in combination with respect to the concentrations required when used separately. CTZ and CFS exhibited synergetic activity against <it>Salmonella </it>by checkerboard microtitre test and the time-kill test. <it>Ex-vivo </it>studies demonstrated a significantly higher intracellular killing of bacteria by macrophages treated with CFS (80 AU/ml) + (CTZ) (2 μg/ml) as compared to when treated with both separately at higher concentrations. Significant reduction in the extent of lipid peroxidation and nitrite levels generated by macrophages in presence of CFS and CTZ, in conjunction, further substantiated the synergistic efficacy of the combination.</p> <p>Conclusions</p> <p>The antimicrobial efficacy of this combination indicates that it may serve as the basis in developing alternative strategies to combat <it>Salmonella </it>infections.</p>