Abstract
The lung in cystic fibrosis (CF) is home to numerous pathogens that shorten the lives of patients. The aim of the present study was to assess changes in the lung bacteriome following antibiotic therapy targeting Pseudomonas aeruginosa in children with CF. The study included nine children (9–18 years) with CF who were treated for their chronic or intermittent positivity for Pseudomonas aeruginosa. The bacteriomes were determined in 16 pairs of sputa collected at the beginning and at the end of a course of intravenous antibiotic therapy via deep sequencing of the variable region 4 of the 16S rRNA gene, and the total bacterial load and selected specific pathogens were assessed using quantitative real-time PCR. The effect of antipseudomonal antibiotics was observable as a profound decrease in the total 16S rDNA load (p = 0.001) as well as in a broad range of individual taxa including Staphylococcus aureus (p = 0.03) and several members of the Streptococcus mitis group (S. oralis, S. mitis, and S. infantis) (p = 0.003). Improvements in forced expiratory volume (FEV1) were associated with an increase in Granulicatella sp. (p = 0.004), whereas a negative association was noted between the total bacterial load and white blood cell count (p = 0.007). In conclusion, the data show how microbial communities differ in reaction to antipseudomonal treatment, suggesting that certain rare species may be associated with clinical parameters. Our work also demonstrates the utility of absolute quantification of bacterial load in addition to the 16S rDNA profiling.
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Acknowledgements
The authors wish to thank Drs. Klára Vilimovská-Dědečková, Veronika Skalicka, Jana Bartosova, and Tereza Jenik. Further, we would like to thank M. Antuskova for technical assistance and A. Bilkova for help with clinical data collection.
Funding
The study was funded by the Grant Agency of Charles University in Prague, grant no. 400213.
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L.K. processed the samples, prepared the libraries, performed sequencing, analyzed the data, and wrote the manuscript; P.D. conceived the study and helped in manuscript writing; J.L. and M.K. performed the statistical analysis; and O.C. supervised the molecular analysis protocol, analyzed the data, and helped in manuscript writing.
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Kramná, L., Dřevínek, P., Lin, J. et al. Changes in the lung bacteriome in relation to antipseudomonal therapy in children with cystic fibrosis. Folia Microbiol 63, 237–248 (2018). https://doi.org/10.1007/s12223-017-0562-3
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DOI: https://doi.org/10.1007/s12223-017-0562-3