Several pasteurization procedures for alfalfa (Medicago sativa) seeds were investigated to completely disinfect inoculated Escherichia coli (Migula) Castellani and Chalmers ATCC(25922). Hot-water treatments (85°C for 9 s) were equally or more effective than 20,000 ppm calcium hypochlorite treatments, yielding a reduction of 2 log₁₀CFU/g. A greater reduction (4 log) was obtained by soaking the seeds prior to the heat treatments and, in some cases, no inoculated bacteria were detected after enrichment of the seeds in peptone. No survival of inoculated bacteria was detected from the seeds or the sprouts after deeply presoaking (15°C for 60 min) and treating the seeds with heat, but the germination decreased to 73% along with the yield which fell to 78.4% that of the control. We describe the hot-water treatment as an alternative method to hypochlorite treatments for disinfecting pathogenic bacteria in seeds for alfalfa sprout production.