Abstract: The process of transport has long been considered an important risk factor for pathogens entry into farms. Disinfection is a generally agreed concept to prevent the introduction of both endemic and epidemic infections, however, potentially toxic, corrosive or volatile problems have arisen because of the use of chemicals as disinfecting agents. Slightly acidic electrolyzed water (SAEW) is considered environmental friendly as it is generated from water and dilute salt solution and reverts to water after use. Also, it has the advantages of possessing broad-spectrum antimicrobial activity, reducing corrosion of surface and minimizing the potential of damage to human health. To reduce the risk of enteric pathogens transmission and leave little residue in animal farms, the disinfection effectiveness of SAEW with pH value of 5.85-6.53 for inactivating Escherichia coli (E. coli) and Salmonella enteritidis (S. enteritidis) mixture on the surface of vehicle tires was evaluated. The coupled effects of tap water washing time (from 2 to 4 min), SAEW treatment time (from 3 to 7 min) and available chlorine concentration (ACC) (from 80 to 140 mg/L) on the reduction of E. coli and S. enteritidis mixture on tires were investigated using a central composite design of the response surface (RS) methodology. The established RS model had a good fitting quantified by determination coefficient (R2) of 0.984 and adjusted determination coefficient of 0.969 (p>0.05). The model was validated with additional random 8 conditions within the experimental domain. The predicted value showed a good agreement with the actual values, for the points of response values were very close to the line of 100% correlation. The results showed that the cleaning time, disinfection time and ACC significantly affected the pathogens reduction (p<0.0001), and their influences were ranked as ACC>disinfection time>cleaning time. The linear correlation coefficients, the quadratic term coefficients and the cross validation coefficients between cleaning time and disinfection time, cleaning time and ACC were significant (p<0.05). The other term coefficient between ACC and disinfection time was not significant. The more reduction of pathogens and the significant interactions between cleaning time and other factors were likely due to the livestock manure, which was a strong limiting factor for disinfection of SAEW. Several authors have stated that the organic soiling could change the formation of combined available chlorines to affect the disinfection effectiveness of SAEW. Therefore, if livestock manure could be more removed by more cleaning time along with more SAEW treatment time and higher ACC, a more effective disinfection would be obtained. Therefore, cleaning time is very important for SAEW disinfection when organic matters exist. The maximum reduction of 1.38 log10cfu/cm2 (92.9%) for E. coli and S. enteritidis mixture was obtained for the vehicle tire washed with tap water for 4 min followed by SAEW treatment for 5 min at an ACC of 140 mg/L. The established RS model could be used. The result proves the potential of the SAEW in disinfection of bacterial cells on tires and in promoting the implementation of disinfection measures to control and reduce the transmission risk of the disease.