A novel virulence plasmid encoding yersiniabactin, salmochelin, and RmpADC from hypervirulent Klebsiella pneumoniae of distinct genetic backgrounds

Hypervirulent Klebsiella pneumoniae (hvKP) is increasingly reported worldwide as a major clinical and public health threat. The virulence of hvKP is attributed largely to the carriage of virulence plasmids (KpVPs). To date, two dominant types of KpVP have been identified, namely, KpVP-1 and KpVP-2. In this study, we reported two hvKP strains from bloodstream infections that carry highly identical virulence plasmids that exhibited <40% coverage compared with KpVP-1 and KpVP-2. This novel virulence plasmid was designated KpVP-3. The two hvKP have different genetic backgrounds, which belonged to ST29-K54 and ST111-K63, respectively. They were both positive for the string test, highly virulent on the Galleria mellonella infection model, and possess high-level macrophage-killing resistance in vitro. Apart from the intrinsic non-susceptibility to ampicillin, both strains were susceptible to commonly used antibiotics. The virulence plasmid carried virulence genes rmpADC, iroBCDN (iro1), and the ybt locus (ybt4) which was not present on either KpVP-1 or KpVP-2. It did not carry antimicrobial resistance genes but carried an incomplete conjugation machinery containing only the traH and traF genes. The KpVP-3 plasmid was stably maintained in both hvKP strains and could not be eliminated with SDS treatment or by serial passage on stress-free agar plates. KpVP-3 was non-self-transmissible under experimental conditions. Data mining suggested KpVP-3-type plasmids have emerged in different countries including China, Australia, and the USA. The emergence of this novel virulence plasmid might pose a potential threat to public health. Heightened efforts are required to study its dissemination mechanism.