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To investigate tetracycline resistance and resistant genotype in  Riemerella anatipestifer , the tetracycline susceptibility of 212  R. anatipestifer  isolates from China between 2011 and 2017 was tested. The results showed that 192 of 212 (90.6%)  R. anatipestifer  isolates exhibited resistance to tetracycline (the MICs ranged from 4 to 256 μg/ml). The results of PCR detection showed that, 170 of 212 (80.2%)  R. anatipestifer  isolates possessed the  tet (X) gene. Other genes, including  tet (A),  tet (M),  tet (Q),  tet (O),  tet (B), and  tet (O/W/32/O), were found at frequencies of 20.8, 4.7, 1.4, 0.9, 0.9, and 0.5%, respectively. However,  tet (C),  tet (E),  tet (G),  tet (K), and  tet (W) were not detected in any isolate. In these  tet  gene positive strains, 31 (14.6%), 2 (0.9%), 5 (2.4%), 1 (0.5%), 3 (1.4%) were detected containing  tet (A)/ tet (X),  tet (M)/ tet (O),  tet (M)/ tet (X),  tet (O)/ tet (X), and  tet (Q)/ tet (X) simultaneously, respectively. One isolates, R131, unexpectedly contained three  tet  genes, i.e.,  tet (M),  tet (O), and  tet (X). Sequence analysis of the  tet  gene ORFs cloned from  R. anatipestifer  isolates confirmed that  tet (A),  tet (B),  tet (M),  tet (O),  tet (Q) and an unusual mosaic  tet  gene  tet (O/W/32/O) were present in  R. anatipestifer . The MIC results of  R. anatipestifer  ATCC 11845 transconjugants carrying  tet (A),  tet (B),  tet (M),  tet (O),  tet (O/W/32/O),  tet (Q), and  tet (X) genes exhibited tetracycline resistance with MIC values ranging from 4 to 64 μg/ml. Additionally, the  tet (X) gene could transfer into susceptible strain via natural transformation (transformation frequencies of ~10 −6 ). In conclusion, the  tet (A),  tet (B),  tet (M),  tet (O),  tet (O/W/32/O),  tet (Q), and  tet (X) genes were found and conferred tetracycline resistance in  R. anatipestifer  isolates. Moreover, the  tet (X) is the main mechanism of tetracycline resistance in  R. anatipestifer  isolates. To our knowledge, this is the first report of  tet (A),  tet (B),  tet (M),  tet (O),  tet (Q), and mosaic gene  tet (O/W/32/O) in  R. anatipestifer .

Various Profiles of tet Genes Addition to tet(X) in Riemerella anatipestifer Isolates From Ducks in China