Omadacycline and eravacycline are gradually being used as new tetracycline antibiotics for the clinical treatment of Gram-negative pathogens. Affected by various tetracycline-inactivating enzymes, there have been reports of resistance to eravacycline and omadacycline in recent years. We isolated a strain carrying the mobile tigecycline resistance gene
tet
(X4) from the feces of a patient in Zhejiang Province, China. The strain belongs to the rare ST485 sequence type. The isolate was identified as
Klebsiella pneumoniae
by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The MICs of antimicrobial agents were determined using either the agar dilution method or the micro broth dilution method. The result showed that the isolate was resistant to eravacycline (MIC = 32 mg/L), omadacycline (MIC > 64 mg/L), and tigecycline (MIC > 32 mg/L). Whole-genome sequencing revealed that the
tet
(X4) resistance gene is located on the IncFII(pCRY) conjugative plasmid.
tet
(X4) is flanked by IS
Vsa3
, and we hypothesize that this association contributes to the spread of the resistance gene. Plasmids were analyzed by S1-nuclease pulsed-field gel electrophoresis (S1-PFGE), Southern blotting, and electrotransformation experiment. We successfully transferred the plasmid carrying
tet
(X4) to the recipient bacteria by electrotransformation experiment. Compared with the DH-5α, the MICs of the transformant L3995-DH5α were increased by eight-fold for eravacycline and two-fold higher for omadacycline. Overall, the emergence of plasmid-borne
tet
(X4) resistance gene in a clinical isolate of
K. pneumoniae
ST485 underscores the essential requirement for the ongoing monitoring of
tet
(X4) to prevent and control its further dissemination in China.
IMPORTANCE
There are still limited reports on
Klebsiella pneumoniae
strains harboring tetracycline-resistant genes in China, and
K. pneumoniae
L3995hy adds a new example to those positive for the
tet
(X4) gene. Importantly, our study raises concerns that plasmid-mediated resistance to omadacycline and eravacycline may spread further to a variety of ecological and clinical pathogens, limiting the choice of medication for extensively drug-resistant bacterial infections. Therefore, it is important to continue to monitor the prevalence and spread of
tet
(X4) and other tetracyclines resistance genes in
K. pneumoniae
and diverse bacterial populations.