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. 2022 Jan 4:15:1-12.
doi: 10.2147/IDR.S343292. eCollection 2022.

Two Phenotypes of Klebsiella pneumoniae ST147 Outbreak from Neonatal Sepsis with a Slight Increase in Virulence

Hua Zou #  1 , Yan Shen #  1 , Chunli Li  1 , Qiuhong Li  1
Affiliations

Two Phenotypes of Klebsiella pneumoniae ST147 Outbreak from Neonatal Sepsis with a Slight Increase in Virulence

Hua Zou et al. Infect Drug Resist. .

Abstract

Purpose: Severe infection has been the leading causes of neonatal death, especially the emergency of multidrug-resistant bacteria such as carbapenem-resistant Enterobacteriaceae. This study aimed to investigate the outbreak of carbapenem-resistant Klebsiella pneumoniae (CR-KP) in neonatal wards and to explore the possible pathogenesis of Klebsiella pneumoniae.

Materials and methods: CR-KP were collected from neonatal ward of Chongqing Health Center for Women and Children between 2017 and 2019. Broth microdilution method was used to evaluate the antimicrobial activities in vitro, at the same time, the virulence of the strain was evaluated by in vitro and in vivo experiments. At last, prokaryotic chain specific transcriptome sequencing was conducted to explore the possible pathogenesis of CR-KP.

Results: In this study, a total of 14 carbapenem-resistant-Klebsiella pneumoniae (CR-KP) strains were isolated from Chongqing Health Center for Women and Children, among which all CR-KP isolates were identified as NDM-1-producers. Molecular epidemiological studies revealed ST147 being the most common sequence type (ST). Moreover, we first found two phenotypes of K. pneumoniae with different virulence from the same specimen. Type I, which was a white and sticky colony had a slight increase in virulence with higher biofilm formation, serum resistance and virulence than Type II with gray colony. Compared with the Type II, 10 pathways were obviously changed in Type I especially amino acid metabolism, such as arginine and proline metabolism.

Conclusion: Our findings revealed a new potential threat of NDM-1-positive CR-KP with higher virulence in neonatal ICU ward. We found two phenotypes of K. pneumoniae with different virulent, which may be due to the difference expression of arginine and proline metabolism.

Keywords: NDM-1; carbapenem-resistant-Klebsiella pneumoniae; neonates; virulence.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
PFGE dendrogram of CR-KP isolates from March, 2017 to February, 2019. All the strains were divided into three clusters. Cluster A belongs to ST147, Clusters B belongs to 3444 and Clusters C belongs to ST17.
Figure 2
Figure 2
Biofilm formation and serum complement-mediated killing of selected CR-KP isolates. (A) Biofilm formation of CR-KP 01 belongs to ST17, CR-KP 14 belongs to ST147 and CR-KP 02 belongs to ST3444. (B) Serum complement-mediated killing of selected CR-KP isolates.
Figure 3
Figure 3
Virulence potential of K. pneumoniae strains in a Galleria mellonella infection model. (A) The survival rate of G. mellonella infected with CR-KP 01inoculum of 1×108 CFU/mL, 1×106 CFU/mL and 1×104 CFU/mL. (B) The survival rate of G. mellonella infected with CR-KP 14 inoculum of 1×108 CFU/mL, 1×106 CFU/mL and 1×104 CFU/mL. (C) The survival rate of G. mellonella infected with CR-KP 02 inoculum of 1×108 CFU/mL, 1×106 CFU/mL and 1×104 CFU/mL.
Figure 4
Figure 4
The characteristics of type I and Type II. (A) PFGE dendrogram of these two types. (B) Biofilm formation of type I and Type II. (C) Serum complement-mediated killing of these two types. (D) Virulence potential of type I and Type II in a Galleria mellonella infection model.
Figure 5
Figure 5
The transcriptomes of type I and Type II. (A) Histogram presentation of GO classification. M: Cell wall/membrane/envelope biogenesis; N: Cell motility; O: Posttranslational modification, protein turnover, chaperones; U: Intracellular trafficking, secretion, and vesicular transport; K: Transcription; L: Replication, recombination and repair; C, Energy production and conversion; E: Amino acid transport and metabolism; F: Nucleotide transport and metabolism; G: Carbohydrate transport and metabolism; H: Coenzyme transport and metabolism; I: Lipid transport and metabolism; P: Inorganic ion transport and metabolism; Q: Secondary metabolites biosynthesis, transport and catabolism; S: Function unknown; The y-axis indicates the number of genes in each category. (B) COG functional classification of type I and Type II.
Figure 6
Figure 6
KEGG pathway analysis of type I and Type II. A total of 115 DEGs were assigned to 41 KEGG pathways. KEGG pathway analysis showed that 10 pathways were obviously changed (P-value <0.05) in theType I. Arginine and proline metabolism, fatty acid degradation, geraniol degradation and phosphotransferase system were overexpressed in type I.
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