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.
Objective: Acinetobacter baumannii is an opportunistic nosocomial pathogen that has increasingly become resistant to carbapenems worldwide. In the Philippines, rates of carbapenem resistance and multidrug resistance are above 50%. We undertook a genomic study of carbapenem-resistant A. baumannii in the Philippines to characterize the population diversity and antimicrobial resistance mechanisms. Methods: We sequenced the whole genomes of 117 A. baumannii isolates recovered by 16 hospitals in the Philippines between 2013 and 2014. From the genome sequences, we determined the multilocus sequence type, presence of acquired determinants of antimicrobial resistance and relatedness between isolates. We also compared the phenotypic and genotypic resistance results. Results: Carbapenem resistance was mainly explained by acquisition of the class-D Beta-lactamase gene blaOXA-23. The concordance between phenotypic and genotypic resistance to imipenem was 98.15%, and it was 94.97% overall for the seven antibiotics analysed. Twenty-two different sequence types were identified, including 7 novel types. The population was dominated by the high-risk international clone 2 (i.e. clonal complex 92), in particular by ST195 and ST208 and their single locus variants. Using whole-genome sequencing, we identified local clusters representing potentially undetected nosocomial outbreaks, as well as multi-hospital clusters that indicated interhospital dissemination. Comparison with global genomes suggested that the establishment of carbapenem-resistant international clone 2 in the Philippines is likely the result of clonal expansion and geographical dissemination, and at least partly explained by inadequate hospital infection control and prevention. Discussion: This is the first extensive genomic study of carbapenem-resistant A. baumannii in the Philippines, and it underscores the importance of hospital infection control and prevention measures to contain high-risk clones.
Background. Salmonella enterica ser. Typhi and Salmonella enterica ser. Paratyphi are agents of typhoid fever, a severe systemic disease, which remains to be a public health concern in the Philippines. Infection due to non-typhoidal Salmonella (NTS), on the other hand, most often results in a self-limiting acute gastroenteritis but may result in invasive disease in some cases. There is scarcity of information on the Salmonella serotypes in the Philippines which limits understanding of the distribution, transmission and antimicrobial resistance of these bacteria.Objective. This study describes the serotype distribution and antimicrobial resistance of Salmonella in the Philippines over a 15-year period.Methodology. Salmonella isolates were collected through the Philippine Department of Health-Antimicrobial Resistance Surveillance Program (DOH-ARSP) from January 1, 2004 to December 31, 2018. The isolates were serotyped using Sven Gard method for slide agglutination using antigens from Denka Seiken (Japan), and S and A serotest (Thailand). Antigenic formula obtained were classified according to White-Kauffmann-LeMinor scheme. Antimicrobial susceptibility testing for ampicillin, ceftriaxone, cefotaxime, chloramphenicol, ciprofloxacin, and trimethoprim-sulfamethoxazole, were performed using both automated and conventional methods (Kirby Bauer disk diffusion and gradient diffusion method). Antimicrobial susceptibility results were interpreted using Clinical and Laboratory Standards Institute (CLSI) 2018 interpretive criteria (M100Ed28E).Results. A total of 2,387 isolates were collected from human specimens during the 15-year study period. There were 69 serotypes of Salmonella identified with the most common being Salmonella enterica ser. Typhi: n=1895 (79.39%), Salmonella enterica ser. Enteritidis: n=182 (7.62%), Salmonella enterica ser. Typhimurium: n=87 (3.64%), Salmonella enterica ser. Weltevreden: n=24 (1.00%), Salmonella enterica ser. Paratyphi A: n=17 (0.71%), Salmonella enterica ser. Stanley: n=17 (0.71%), Salmonella enterica ser. Anatum: n=13 (0.54%), Salmonella enterica ser. Heidelberg: n=12 (0.50%), Salmonella enterica ser. Choleraesuis var. Kunzendorf: n=9 (0.38%). The multidrug resistant Salmonella serotypes reported in this study were mostly resistant to ampicillin, cefotaxime, ciprofloxacin combinations. Conclusion.This present study showed that prevailing Salmonella serotypes in the Philippines were similar with Salmonella serotypes reported from other Asian countries. Typhoidal isolates were high among 6-17 years old and were mostly from males. The antimicrobial resistance rates for typhoidal Salmonella isolates to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, ciprofloxacin, ceftriaxone and cefotaxime were lower compared with the antimicrobial resistance rates for non-typhoidal Salmonella isolates. Multidrug resistance for both Salmonella Typhi and NTS were relatively low. Continued and enhanced surveillance is needed to monitor the rising levels of antimicrobial resistance, determine risk ...
Pseudomonas aeruginosa is an opportunistic pathogen that often causes nosocomial infections resistant to treatment. Rates of antimicrobial resistance (AMR) are increasing, as are rates of multidrug-resistant (MDR) and possible extensively drug-resistant (XDR) infections. Our objective was to characterize the molecular epidemiology and AMR mechanisms of this pathogen. We sequenced the whole genome for each of 176 P. aeruginosa isolates collected in the Philippines in 2013–2014; derived the multilocus sequence type (MLST), presence of AMR determinants and relatedness between isolates; and determined concordance between phenotypic and genotypic resistance. Carbapenem resistance was associated with loss of function of the OprD porin and acquisition of the metallo-β-lactamase (MBL) gene blaVIM. Concordance between phenotypic and genotypic resistance was 93.27% overall for six antibiotics in three classes, but varied among minoglycosides. The population of P. aeruginosa was diverse, with clonal expansions of XDR genomes belonging to MLSTs ST235, ST244, ST309 and ST773. We found evidence of persistence or reintroduction of the predominant clone ST235 in one hospital, and of transfer between hospitals. Most of the ST235 genomes formed a distinct lineage from global genomes, thus raising the possibility that they may be unique to the Philippines. In addition, long-read sequencing of one representative XDR ST235 isolate identified an integron carrying multiple resistance genes (including blaVIM-2), with differences in gene composition and synteny from the P. aeruginosa class 1 integrons described previously. The survey bridges the gap in genomic data from the Western Pacific Region and will be useful for ongoing surveillance; it also highlights the importance of curtailing the spread of ST235 within the Philippines.
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