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Elizabethkingia anophelis MSU001 Isolated from Anopheles stephensi: Molecular Characterization and Comparative Genome Analysis - PubMed

  • ️Mon Jan 01 2024

Elizabethkingia anophelis MSU001 Isolated from Anopheles stephensi: Molecular Characterization and Comparative Genome Analysis

Shicheng Chen et al. Microorganisms. 2024.

Abstract

Elizabethkingia anophelis MSU001, isolated from Anopheles stephensi in the laboratory, was characterized by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-ToF/MS), biochemical testing, and genome sequencing. Average nucleotide identity analysis revealed 99% identity with the type species E. anophelis R26. Phylogenetic placement showed that it formed a clade with other mosquito-associated strains and departed from a clade of clinical isolates. Comparative genome analyses further showed that it shared at least 98.6% of genes with mosquito-associated isolates (except E. anophelis As1), while it shared at most 88.8% of common genes with clinical isolates. Metabolites from MSU001 significantly inhibited growth of E. coli but not the mosquito gut symbionts Serratia marcescens and Asaia sp. W12. Insect-associated E. anophelis carried unique glycoside hydrolase (GH) and auxiliary activities (AAs) encoding genes distinct from those of clinical isolates, indicating their potential role in reshaping chitin structure and other components involved in larval development or formation of the peritrophic matrix. Like other Elizabethkingia, MSU001 also carried abundant genes encoding two-component system proteins (51), transcription factor proteins (188), and DNA-binding proteins (13). E. anophelis MSU001 contains a repertoire of antibiotic resistance genes and several virulence factors. Its potential for opportunistic infections in humans should be further evaluated prior to implementation as a paratransgenesis agent (by transgenesis of a symbiont of the vector).

Keywords: Elizabethkingia anopheles; genome analysis; interaction; symbiotic traits.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1

Growth features and microscopic observation of E. anophelis MSU001. (A) Hemolytic activity on sheep blood agar; (B) motility test; (C) scan electron microscopy; (D) demonstration of bacterial morphology by electron microscopy with negative stain.

Figure 2
Figure 2

Phylogenetic placement of E. anophelis MSU001. The tree was constructed with 18 genomes with a core of 2307 genes per genome, 41,526 in total. The core had 783,693 amino acid residues/bp per genome, 14,106,474 in total. The horizontal bar represents 0.05 substitutions per site.

Figure 3
Figure 3

Pan and core genome evolution according to the number of selected Elizabethkingia genomes. (A) Number of genes (pan-genome) for a given number of sequentially added genomes. A pan development plot was generated for the following genomes: E. anophelis Ag1 (NZ_CP023402), E. anophelis R26 (NZ_CP023401), E. anophelis 2_62 (NZ_CP071551), E. anophelis 296_96 (NZ_CP046080), E. anophelis AR4_6 (NZ_CP023404), E. anophelis AR6_8 (NZ_CP023403), E. anophelis As1 (NZ_LFKT01000002), E. anophelis CSID_3000521207 (NZ_CP015067), E. anophelis F3201 (NZ_CP016375), E. anophelis JM_87 (NZ_CP016372), E. anophelis MSU001 (NZ_JAHDTL010000009), E. anophelis SUE (NZ_CP034247), E. anophelis LDVH-AR107 (NZ CP023403), E. anophelis JUNP 353 (NZ_ AP022313). (B) Number of shared genes (core genome) as a function of the number of genomes sequentially added. The genomes used for generating the core genome development plot were the same as listed in (A).

Figure 4
Figure 4

Venn diagram illustrating the distribution of shared and specific clusters of orthologous groups in the selected Elizabethkingia genomes. (A) Venn diagram of shared and unique genes in the selected mosquito-associated Elizabethkiniga. (B) Venn diagram of shared and unique genes in MSU001 and the clinically important Elizabethkiniga. The unique and shared genomes among the compared genomes were determined using the BLAST score ratio approach of EDGAR 3.2 with a cutoff of 30%.

Figure 5
Figure 5

Inhibitory effects of Elizabethkingia metabolites on selected bacteria. * Statistically significant difference (p < 0.05). (A) Growth comparison between wild type strain for arginine utilization (SCH814) and arginine metabolism mutant (SCH873) in the M9 medium and M9 medium supplemented with 20-fold diluted LB medium. (B) Comparison between growth of SCH814 and SCH873 in mosquitoes. (C) The effects of spent media on the growth of Asaia sp. W12, Serratia marcescens and E. coli. The spent broth from E. anophelis MSU001 (48-h incubation) was added E. coli, Serratia marcescens ano1 and Asaia sp. W12, statically cultured at 28 °C for 24 h and plated on their respective solid agar media for CFU calculation.

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References

    1. Coyle A.L. Elizabethkingia anophelis: Exploring the outbreak of disease in the Midwest. Nursing. 2017;47:61–63. doi: 10.1097/01.NURSE.0000512887.67622.84. - DOI - PubMed
    1. Janda J.M., Lopez D.L. Mini Review: New Pathogen Profiles: Elizabethkingia anophelis. Diagn. Microbiol. Infect. Dis. 2017;88:201–205. doi: 10.1016/j.diagmicrobio.2017.03.007. - DOI - PubMed
    1. Kämpfer P., Matthews H., Glaeser S.P., Martin K., Lodders N., Faye I. Elizabethkingia anophelis Sp. Nov., Isolated from the Midgut of the Mosquito Anopheles gambiae. Int. J. Syst. Evol. Microbiol. 2011;61:2670–2675. doi: 10.1099/ijs.0.026393-0. - DOI - PubMed
    1. Breurec S., Criscuolo A., Diancourt L., Rendueles O., Vandenbogaert M., Passet V., Caro V., Rocha E.P.C., Touchon M., Brisse S. Genomic Epidemiology and Global Diversity of the Emerging Bacterial Pathogen Elizabethkingia anophelis. Sci. Rep. 2016;6:30379. doi: 10.1038/srep30379. - DOI - PMC - PubMed
    1. Ganley J.G., D’Ambrosio H.K., Shieh M., Derbyshire E.R. Coculturing of Mosquito-Microbiome Bacteria Promotes Heme Degradation in Elizabethkingia anophelis. ChemBioChem. 2020;21:1279–1284. doi: 10.1002/cbic.201900675. - DOI - PubMed

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