Bacterial resistance to the third-generation cephalosporin antibiotics has become a major concern for public health. This study was aimed to determine the characteristics and distribution of bla(CTX-M-14), which encodes an extended-spectrum beta-lactamase, in Escherichia coli isolated from Guangdong Province, China. A total of 979 E. coli isolates isolated from healthy or diseased food-producing animals including swine and avian were examined for bla(CTX-M-14) and then the bla(CTX-M-14_)positive isolates were detected by other resistance determinants [extended-spectrum beta-lactamase genes, plasmid-mediated quinolone resistance, rmtB, and floR] and analyzed by phylogenetic grouping analysis, PCR-based plasmid replicon typing, multilocus sequence typing, and plasmid analysis. The genetic environments of bla(CTX-M-14) were also determined by PCR. The results showed that fourteen CTX-M-14-producing E. coli were identified, belonging to groups A (7/14), B1 (4/14), and D (3/14). The most predominant resistance gene was bla(TEM) (n = 8), followed by floR (n = 7), oqxA (n = 3), aac(6')-1b-cr (n = 2), and rmtB (n = 1). Plasmids carrying bla(CTX-M-14) were classified to IncK, IncHI2, IncHI1, IncN, IncFIB, IncF or Inch, ranged from about 30 to 200 kb, and with insertion sequence of ISEcp1, IS26, or ORF513 located upstream and IS903 downstream of bla(CTX-M-14). The result of multilocus sequence typing showed that 14 isolates had 11 STs, and the 11 STs belonged to five groups. Many of the identified sequence types are reported to be common in E. coli isolates associated with extraintestinal infections in humans, suggesting possible transmission of bla(CTX-M-14) between animals and humans. The difference in the flanking sequences of bla(CTX-M-14) between the 2009 isolates and the early ones suggests that the resistance gene context continues to evolve in E. coli of food producing animals.