The increase of Salmonella enterica strains showing resistance against antibiotics has resulted in limiting the effective treatment of human infections. The present study characterized the resistance to tetracycline in S. enterica serovar Typhimurium strains, recovered from irrigation water in distinct regions in the Culiacan Valley, an important agricultural region in Mexico for horticultural crops that are exported to the United States. Analysis of the genomic diversity by pulse-field gel electrophoresis (PFGE) typing showed that the Salmonella Typhimurium strains were grouped into four distinct genotypic clusters, indicating genomic diversity among 12 strains examined. The polymerase chain reaction and DNA sequencing analysis demonstrated that the tet(A) gene was found on the genomic DNA and was located within a truncated version of transposon Tn1721. The comparative analysis of the tet(A) gene sequence in Salmonella Typhimurium strains identified high sequence similarity to the tet determinant of plasmid RP1, which is homologous to the tet gene in Tn1721. The findings show the presence of tet(A) among the tetracycline-resistant Salmonella Typhimurium strains isolated from irrigation water used for growing fresh fruits and vegetables.