Using next generation sequencing approaches to define the population biology of the neglected cystic fibrosis lung pathogen Burkholderia multivorans

Parfitt, Kasia 2022. Using next generation sequencing approaches to define the population biology of the neglected cystic fibrosis lung pathogen Burkholderia multivorans. PhD Thesis, Cardiff University. Item availability restricted.

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Abstract

Burkholderia multivorans is the most frequently isolated Burkholderia cepacia complex species recovered from cystic fibrosis lung infection. However, its pathogenesis and species population biology remain elusive. Understanding adaptational factors of B. multivorans to the CF lung microenvironment is important for predicting its pathogenesis and disease outcome. B. multivorans population biology was explored using pan genome analysis, average nucleotide identity and phylogenomic analysis (n = 283). The population split into two major genomic lineages, designated 1 and 2, and four B. multivorans model strains were selected to represent them: the soil strain ATCC 17616 (lineage 2a), BCC1272 (lineage 2a), BCC0033 (lineage 2b), and BCC0084 (lineage 1). The latter 3 CF strains were completely genome sequenced to add to the readily available reference genome ATCC 17616. Using gene presence-absence analysis, unique B. multivorans lineage-specific genes were identified. This enabled diagnostic PCR design with genes ghrB_1 and glnM_2 selected as the lineage 1 and lineage 2 targets, respectively. The PCRs showed 100% lineage-specificity against 48 B. multivorans strains. Phenotypic analysis was performed on a subset of 49 B. multivorans strains evaluating their morphology, growth kinetics, motility, biofilm formation, and exopolysaccharide production. The B. multivorans phenotype was variable amongst the strains, with no link to genomic lineage. Phenotypic comparison was also performed when B. multivorans were mixed with a secondary CF pathogen. The suppression of P. aeruginosa LESB58 protease production, when cultured with B. multivorans, was identified as an interesting interaction based on an unknown mechanism. Three of the B. multivorans model strains (BCC0033, BCC0084, and ATCC 17616) were also evaluated in a murine respiratory infection model and all showed good persistence over 5-days. Overall, this work has built a foundation of knowledge on the B. multivorans phenotype and genotype, enabling associations between lineage, therapeutics testing, and clinical outcome to be studied

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Biosciences
Subjects:	Q Science > Q Science (General)
Date of First Compliant Deposit:	24 November 2022
Date of Acceptance:	24 November 2022
Last Modified:	24 Nov 2022 16:28
URI:	https://orca.cardiff.ac.uk/id/eprint/154453

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