Global surveillance of AMR through sewage samples and ... Pimlapas... · Global surveillance of...
Transcript of Global surveillance of AMR through sewage samples and ... Pimlapas... · Global surveillance of...
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Global surveillance of antimicrobial resistance through global sewage
Pimlapas Leekitcharoenphon (Shinny)Research Group for Genomic Epidemiology
DTU FoodTechnical University of Denmark (DTU)
@ShinnyPimlapas
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Global surveillance of AMR
Can human sewage be used to detect and combined with modelling explain global emergence and trends in AMR ?
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79 samples from 60 countries have been collected and analysed
Sample collection - 2016
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https://cge.cbs.dtu.dk/services/ResFinder/
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https://cge.cbs.dtu.dk/services/MGmapper/
https://bitbucket.org/genomicepidemiology/mgmapper
https://www.coursera.org/learn/metagenomics
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mapping AMR result
FPKM
Fragments Per Kilobase reference per Million bacterial fragments
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Total FPKMFragments Per Kilobase reference per Million bacterial fragments
Hendriksen RS, et al. Nature Communications, 2019
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AMR classes
Hendriksen RS, et al. Nature Communications, 2019
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AMR genes
Hendriksen RS, et al. Nature Communications, 2019
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Resistome clustering in sewage across regionsHellinger-transformed (decostand function in vegan package) and Bray-Curtis dissimilarity
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AMR class
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Drivers for AMR
Factor Significance
Temperature -
Flight connections -
Antimicrobial use *
Human development index ***
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Predict AMR level using socio-economic data
AMR abundance
Predicted AMR
abundance
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Global resistance prediction
Hendriksen RS, et al. Nature Communications, 2019
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Denmark
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Conclusion
• Sewage is a good source for an ethically acceptable and economically feasible global surveillance and prediction of AMR and infectious diseases
• Drug use explains only minor part of AMR • There are other factors that drive the occurrence
of AMR such as sanitation, health …
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Acknowledgement• Technical University of Denmark (DTU)
– Frank Aarestrup– Rene Hendriksen– Thomas Nordahl Petersen– Patrick Munk– Sunje J. Pamp– Patrick Njage– Timo Roder– Jette Kjeldgaard
• University of Edinburgh – Bram Van Bunnik– Mark Woolhouse
• Erasmus Medical Center
- Marion P. Koopmans
- David Nieuwenhuijse
• Global sewage surveillance project consortium21
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Thank you
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Conclusion
• Sewage is a good source for an ethically acceptable and economically feasible global surveillance and prediction of AMR and infectious diseases
• AMR abundance is mainly influenced by local/national parameters
• Drug use explains only minor part of AMR • There are other factors that drive the occurrence
of AMR such as sanitation, health …
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Sample collection protocol
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Packing instruction
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Sample details
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Sample collection – 2016 (pilot)
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AMR class
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Top 50 functional resistance genes
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Functional resistance• Resistance phenotype based on a functional
metagenomic antibiotic resistance determinants• Four functional metagenomic studies were
considered, that analysed human faecal samples, sewage, latrines and soils
• Only ARDs selected on antibiotics towards which they are assumed to confer resistance have been selected
• Homology reduction with ResFinder using cd-hit1. Pehrsson, E. C. et al. Interconnected microbiomes and resistomes in low-income human habitats. Nature 533, 212Ð216 (2016).2. Moore, A. M. et al. Pediatric fecal microbiota harbor diverse and novel antibiotic resistance genes. PloS One 8, e78822 (2013).
3. Sommer, M. O. A., Dantas, G. & Church, G. M. Functional characterization of the antibiotic resistance reservoir in the human microflora. Science 325, 1128Ð1131 (2009).4. Forsberg, K. J. et al. Bacterial phylogeny structures soil resistomes across habitats. Nature 509, 612Ð616 (2014).
5. Seemann, T. Prokka: rapid prokaryotic genome annotation. Bioinforma. Oxf. Engl. 30, 2068Ð2069 (2014).6. Fu, L., Niu, B., Zhu, Z., Wu, S. & Li, W. CD-HIT: accelerated for clustering the next-generation sequencing data. Bioinforma. Oxf. Engl. 28, 3150Ð3152 (2012).
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Moore AM1, Munck C, Sommer MO, Dantas G. Front Microbiol. 2011 Oct 17;2:188
Functional metagenomics
faecalsewage
toiletsoils
1. Pehrsson, E. C. et al. Interconnected microbiomes and resistomes in low-income human habitats. Nature 533, 212Ð216 (2016).2. Moore, A. M. et al. Pediatric fecal microbiota harbor diverse and novel antibiotic resistance genes. PloS One 8, e78822 (2013).
3. Sommer, M. O. A., Dantas, G. & Church, G. M. Functional characterization of the antibiotic resistance reservoir in the human microflora. Science 325, 1128Ð1131 (2009).4. Forsberg, K. J. et al. Bacterial phylogeny structures soil resistomes across habitats. Nature 509, 612Ð616 (2014).
5. Seemann, T. Prokka: rapid prokaryotic genome annotation. Bioinforma. Oxf. Engl. 30, 2068Ð2069 (2014).6. Fu, L., Niu, B., Zhu, Z., Wu, S. & Li, W. CD-HIT: accelerated for clustering the next-generation sequencing data. Bioinforma. Oxf. Engl. 28, 3150Ð3152 (2012).
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https://cge.cbs.dtu.dk/services/ResFinderFG/
ResFinder -> 3026 genes
Functional resistance -> 2514 genes
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Pig Poultry
Munk P. et al. Nature Microbiology. 898–908 (2018)
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aminoglycoside
aminoglycoside;quinolone
beta-lactam
colistin
fosfomycin
glycopeptide
macrolide
macrolide;oxazolidinone;phenicol
nitroimidazole
oxazolidinone
oxazolidinone;phenicol
phenicol
quinolone
rifampicin
sulphonamide
tetracycline
trimethoprim
ResFinder Functional resistance
Critically important (High priority)Highly important
Critically important (Highest priority)
Important
aminoglycoside
beta_lactam
cotrimoxazole
D_cycloserine
phenicol
quinolone
tetracycline
trimethoprim
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What are the other drivers of the occurrence of AMR ?
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Future perspective
• Diversity and Abundance –AMR in gene level–AMR organisms
• Other resistance such as metal resistance and chromosomal point mutation
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What are the other drivers of the occurrence of AMR ?
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AMR genes and E.coli association
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R-sq 1.95 %p-value 0.22
R-sq 13.67 %p-value 0.0008
R-sq 8.39 %p-value 0.0096
R-sq 27.59 %p-value 6.708e-07
ResFinder (total FPKM) colistin
rifampicinquinolone
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R-sq 6.01 %p-value 0.029
R-sq 35.80 %p-value 5.768e-09
R-sq 10.41 %p-value 0.0037
R-sq 10.74 %p-value 0.0032
Functional resistance (total FPKM)D-cycloserine
cotrimoxazole
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R-sq 21.49 %p-value 1.692e-05
trimethoprim
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