National networks of laboratory-based surveillance of antimicrobial resistance (AMR) monitor resistance trends and disseminate these data to AMR stakeholders. Whole-genome sequencing (WGS) can support surveillance by pinpointing resistance mechanisms and uncovering transmission patterns. However, genomic surveillance is rare in low-and middleincome countries. Here, we implement WGS within the established Antimicrobial Resistance Surveillance Program of the Philippines via a binational collaboration. In parallel, we characterize bacterial populations of key bug-drug combinations via a retrospective sequencing survey. By linking the resistance phenotypes to genomic data, we reveal the interplay of genetic lineages (strains), AMR mechanisms, and AMR vehicles underlying the expansion of specific resistance phenotypes that coincide with the growing carbapenem resistance rates observed since 2010. Our results enhance our understanding of the drivers of carbapenem resistance in the Philippines, while also serving as the genetic background to contextualize ongoing local prospective surveillance.

Background. Drug-resistant bacterial infections constitute a growing threat to public health globally. National networks of laboratory-based surveillance of antimicrobial resistance (AMR) monitor the emergence and spread of resistance and are central to the dissemination of these data to AMR stakeholders. Whole-genome sequencing (WGS) can support these efforts by pinpointing resistance mechanisms and uncovering transmission patterns. We implemented WGS within the established Antimicrobial Resistance Surveillance Program (ARSP) of the Philippines. We aimed to employ WGS to characterize bacterial populations and dissect resistance phenotypes of key bug-drug combinations, thus establishing a genetic background to contextualize local prospective surveillance.Methods. We sequenced the genomes from eight bacterial pathogens collected between 2013 and 2014 by the ARSP, and conducted phylogenetic analyses, in silico genotyping, genomic predictions of AMR, and characterization of key plasmids carrying carbapenemase genes. Here, we focus on carbapenemase-producing organisms.Findings. ARSP phenotypic data indicated increasing carbapenem resistance for Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae and Escherichia coli, with marked expansion of specific resistance phenotypes. By linking resistance phenotypes to genomic data, we revealed the diversity of genetic lineages (strains), AMR mechanisms, and AMR vehicles underlying this expansion. We discovered a previously unidentified plasmid-driven hospital outbreak of carbapenem-resistant K. pneumoniae, uncovered the interplay of carbapenem resistance genes and plasmids in the geographic circulation of international epidemic K. pneumoniae ST147, and found that carbapenem-resistant E. coli ST410 were represented by diverse lineages of global circulation that both conserved international plasmids and acquired plasmids of local circulation.Conclusions. WGS provided an enhanced understanding of the interplay between strains, genes and vehicles driving the dissemination of carbapenem resistance in the Philippines. We generated a blueprint for the integration of WGS and genomic epidemiology into an established national system of laboratory-based surveillance of AMR through international collaboration. Continued prospective sequencing, capacity building and collaboration will strengthen genomic surveillance of AMR in the Philippines and the translation of genomic data into public-health action.

Background Klebsiella pneumoniae is a critically important pathogen in the Philippines. Isolates are commonly resistant to at least 2 classes of antibiotics, yet mechanisms and spread of its resistance are not well studied. Methods A retrospective sequencing survey was performed on carbapenem-, extended spectrum beta-lactam-, and cephalosporin-resistant Klebsiella pneumoniae isolated at 20 antimicrobial resistance (AMR) surveillance sentinel sites from 2015 through 2017. We characterized 259 isolates using biochemical methods, antimicrobial susceptibility testing, and whole-genome sequencing (WGS). Known AMR mechanisms were identified. Potential outbreaks were investigated by detecting clusters from epidemiologic, phenotypic, and genome-derived data. Results Prevalent AMR mechanisms detected include bla CTX-M-15 (76.8%) and bla NDM-1 (37.5%). An epidemic IncFII(Yp) plasmid carrying bla NDM-1 was also detected in 46 isolates from 6 sentinel sites and 14 different sequence types (STs). This plasmid was also identified as the main vehicle of carbapenem resistance in 2 previously unrecognized local outbreaks of ST348 and ST283 at 2 different sentinel sites. A third local outbreak of ST397 was also identified but without the IncFII(Yp) plasmid. Isolates in each outbreak site showed identical STs and K- and O-loci, and similar resistance profiles and AMR genes. All outbreak isolates were collected from blood of children aged < 1 year. Conclusion WGS provided a better understanding of the epidemiology of multidrug resistant Klebsiella in the Philippines, which was not possible with only phenotypic and epidemiologic data. The identification of 3 previously unrecognized Klebsiella outbreaks highlights the utility of WGS in outbreak detection, as well as its importance in public health and in implementing infection control programs.

Objective. Scarcity of data on the serotype composition and antibiotic resistance of invasive pneumococci from developing countries has been noted. 13 We describe in this study the serogroup distribution and antimicrobial resistance patterns of Streptococcus pneumoniae in the Philippines from 2004-2011.Methodology. S. pneumoniae isolated from patients with invasive pneumococcal disease (IPD) were referred to the Antimicrobial Resistance Surveillance Reference Laboratory from 2004 to 2011. Typing of isolates was done through slide agglutination and antimicrobial susceptibility was determined following CLSI methods.Results. The penicillin-resistant meningitis isolates were of serotypes 1, 5 and 14 which are covered by PCV10 and 13. The erythromycin resistant isolates were serotype 9 while cotrimoxazole resistant isolates were serotypes 1, 5, 6, 12 and 14. Forty-one percent of the cotrimoxazole resistant isolates are covered by PCV7, and 88% are covered by both PCV10 and PCV13. Levofloxacin resistant isolates were of serotypes 5 and 23 with PCV7 coverage of 50% and PCV10 and PCV13 coverages of 100%.Conclusions. S. pneumoniae serotypes causing IPD in the country is largely similar to the dominant IPD serotypes worldwide. The serotype distribution in the Philippines remained stable from 2004 to 2011 and antimicrobial resistance among the isolates remained low. The serotypes of antibiotic resistant S. pneumoniae in this study were not similar with known serotype resistance profiles in other Asian countries. With the inclusion of PCV in the free national immunization program of the country beginning 2013, continued surveillance of prevailing pneumococcal serotypes should be done to monitor any shift in the prevalence of PCV associated serotypes to guide disease control measures including control of emergence of resistant pneumococcal isolates.

Performing whole genome sequencing (WGS) for the surveillance of antimicrobial resistance (AMR) offers the ability to determine not only the antimicrobials to which rates of resistance are increasing, but also the evolutionary mechanisms and transmission routes responsible for the increase at local, national, and global scales. To derive WGS-based outputs, a series of processes are required, beginning with sample and metadata collection, followed by nucleic acid extraction, library preparation, sequencing, and analysis. Throughout this pathway there are many data-related operations required (informatics) combined with more biologically-focused procedures (bioinformatics). For a laboratory aiming to implement pathogen genomics, the informatics and bioinformatics activities can be a barrier to starting on the journey; for a laboratory that has already started, these activities may become overwhelming. Here we describe these data bottlenecks and how they have been addressed in laboratories in India, Colombia, Nigeria, and the Philippines, as part of the NIHR Global Health Research Unit on Genomic Surveillance of AMR. The approaches taken include the use of reproducible data parsing pipelines and genome sequence analysis workflows, using technologies such as Data-flo, the Nextflow workflow manager, and containerization of software dependencies. By overcoming barriers to WGS implementation in countries where genome sampling for some species may be underrepresented, a body of evidence can be built to determine the concordance of antimicrobial sensitivity testing (AST) and genome-derived resistance, novel high-risk clones can be discovered, and unknown mechanisms of resistance discovered.