Biology subjects

Corander, J.

Publications and source records attributed to Corander, J..

At least 19 recordsLinked to original sources

bioRxiv2026

Identifying candidate genetic determinants associated with invasion in pneumococcal lineages

Serotype invasiveness estimates help identify high-risk serotypes and interpret disease replacement following pneumococcal conjugate vaccination. Because serotype and lineage are strongly associated, their independent contributions must be separated to identify unexpectedly invasive lineages and investigate underlying mechanisms. Using temporally matched carriage and invasive-disease isolates from children aged <7 years in South Africa and the USA, we fitted a logistic mixed-effects model with Global Pneumococcal Sequence Cluster (GPSC) as a random effect and serotype and country as fixed effects. We conducted a species-wide genome-wide association study (GWAS) of invasion and compared the prevalence of significant unitigs in GPSC1 and GPSC21 to identify invasion-associated variants differentiating them. We additionally examined capsule-locus variation that could contribute to their contrasting invasiveness. After serotype and country adjustment, GPSC1 and GPSC21, which both express 19F and share an immediate common ancestor, ranked fifth and 149th of 150 lineages; odds multipliers were 1.51 (95% conditional interval 1.06-2.15) and 0.61 (0.41-0.93), respectively. Of 1,569 invasion-associated unitigs from the species-wide GWAS, 272 differentiated GPSC1 from GPSC21. Among these, the strongest signal mapped to hk11 (desK) and was associated with GPSC1; bgaA was the highest-ranked known antibody-binding target. Within GPSC1 CC271 had additional variants in the ATP-dependent Clp protease ATP-binding subunit ClpX compared to the CC420 subclade. Capsule-protein folds were conserved with local effects indistinguishable from prediction uncertainty. Separating serotype and lineage effects can identify unexpectedly invasive lineages and prioritize candidate serotype-independent determinants, vaccine antigens and capsule variation for experimental investigation.

microbiology↗

bioRxiv2026

Rapid gene exchange explains differences in bacterial pangenome structure

The size and diversity of bacterial gene repertoires, known as pangenomes, vary widely across species. The evolutionary forces driving the maintenance of pangenomes is an open topic of debate, with contradictory theories suggesting that pangenomes exist as a result of neutral evolution, with all genes gained and lost at random, or that all genes provide a fitness benefit to the host and are maintained by positive selection. Modelling of pangenome dynamics has provided insight into how gene exchange explains observed gene frequency distributions, and stands as the only means of jointly inferring contributions of individual gene selection effects and mobility on the maintenance of pangenomes. However, previous modelling studies have not included both gene-level selection and mobility, and do not consider broadly sampled genome datasets for many species. To differentiate neutral and selective forces maintaining pangenomes, we developed a mechanistic model of gene-level evolution, Pansim, and a scalable model fitting framework, PopPUNK-mod. Together, these tools leverage rapid genome distance calculation to fit models of pangenome dynamics to datasets containing hundreds of thousands of genomes. We used this framework to compare the pangenome dynamics of over 400 different bacterial species, using over 600,000 genomes. We find that diversity in pangenome characteristics between species is driven predominantly by variation in the number of rapidly exchanged genes, while the rate of exchange of remaining genes is conserved. We find that bacterial phylogeny, rather than ecology, correlates with pangenome dynamics. We express that pan-species gene-level analyses are now needed to understand selection across accessory genes. Our work highlights the importance of gene exchange rate differences in governing differences in pangenome characteristics between species.

bioinformatics↗

bioRxiv2026

A 28-year evolution experiment on Burkholderia pseudomallei survival in nutrient-depleted sterile water

Environmental persistence allows opportunistic pathogens to survive a range of harsh conditions, increasing the likelihood of eventual infection. Burkholderia pseudomallei, the causative agent of melioidosis, can endure long-term nutrient-depletion in the environment, but its adaptive mechanisms remain poorly understood. Here, we investigated the evolutionary trajectory of a clinical B. pseudomallei strain maintained in sterile water since 1994. The strain was inoculated into nine individual tubes at nine initial concentrations (102-1010 CFU/mL) and has remained viable to date. Liquid chromatography-mass spectrometry analysis of the water identified potential carbon sources, including phthalic acid -- a plastic degradation product likely leached from inoculation tubes -- which the strain can metabolise via an intact catabolic operon. Genomic variations accumulated between 1994 and 2022 were characterised using both single-colony and plate-sweep sequencing which provided complementary insights. Across all tubes, we identified 249 single-nucleotide polymorphisms (SNPs), 73 indels, and a large-scale deletion. Cultures from each tube displayed a consistently low mutation rate (3.18 x 10- SNPs per site per year), suggesting that cells entered a dormant or slow-growth state. Of 393 genes with mutations, 193 were independently mutated in more than one tube, particularly those involved in signal transduction, cell wall and membrane biogenesis, and secondary metabolite synthesis. These patterns indicate parallel adaptation to long-term nutrient deprivation through modulation of cell-density-related functions and loss of metabolically costly pathways. Remarkably, B. pseudomallei from this experiment remain viable after nearly three decades, providing a rare natural model for understanding how environmental bacteria endure and adapt in extremely nutrient-depleted conditions.

evolutionary biology↗

bioRxiv2025

Mapping the evolutionary path towards multi-drug resistance in the pandemic Escherichia coli ST131 lineage

Escherichia coli sequence type (ST) 131 is the most widely studied genetic lineage of the species to date, originally identified in the early 2000s as an increasingly common cause of human urinary tract and bloodstream infections worldwide. This lineage is subdivided into four extant main subclades A, B, C1 and C2 that exhibit distinct features in terms of invasiveness, antibiotic resistance and transmissibility. However, the evolutionary pathway from the generally susceptible ST131-B to the drug-resistant ST131-C clades remains poorly mapped. To fill this knowledge gap, we analysed in detail human clinical isolates obtained in Vietnam, designated as belonging to the generally neglected minor clade ST131-B0. We sequenced them using both short- and long-read technology, and combined these data with a recently published high-resolution genomic collection to provide further insight into the evolutionary process and its timeline. Extensive genomic analyses established ST131-B0 as an intermediary progenitor in the evolutionary path leading from ST131-B to the ST131-C clades, associated with multiple type I pili switches, as well as the loss and gain of specific chromosomal genes representing diverse core functions such as metabolism, transcription, DNA binding and type II toxin-antitoxin systems. Furthermore, all Vietnamese isolates of ST131-B0 unprecedentedly harboured blaCTX-M genes encoding extended-spectrum {beta}-lactamases, a trait dominant in ST131-C clades and not previously observed in ST131-B0. Our study supports the hypothesis that the ST131-C clades have gradually evolved from ST131-B by reducing the host range with better adaptation to colonising humans under selective conditions.

evolutionary biology↗

bioRxiv2025

Co-evolution between colibactin production and resistance is linked to clonal expansions in Escherichia coli

Specific strains of Escherichia coli employ the polyketide synthase island to produce a metabolite called colibactin that is implicated in colorectal tumorigenesis via its genotoxic effect on human DNA. However, the damage to host DNA is essentially collateral from ecological competition during bacterial colonisation, where colibactin is used to displace other gut bacteria. Despite intensive recent research on colibactin and its involvement in tumorigenesis, the evolutionary dynamics related to its production remain poorly characterised. We map evolution of colibactin production in E. coli using a large collection of high-resolution genomes and show that its introduction via a mobile element has induced multiple plasmid-mediated acquisitions of previously unrecognised colibactin immunity genes in non-producing multi-drug resistant lineages. Our study suggests that in E. coli, multi-drug resistance is incompatible with colibactin production due to the unbearable combined fitness costs, supported by colibactin-producing phenotypes remaining susceptible to most classes of antibiotics and thriving in regions with low antimicrobial usage. Consequently, resistant lineages circulating in these regions are under selective pressure to acquire colibactin immunity to compete with the endemic colibactin-producing bacteria during colonisation. Conversely, high antimicrobial usage selects for a colibactin susceptible antimicrobial resistant phenotype as it drives the colibactin-producing E. coli towards extinction. Such a duality of evolutionary strategies to become endemic in a host population structured by varying ecological pressures may hold more generally across bacterial species colonizing the human gut.

microbiology↗

bioRxiv2025

Megaplasmids associate with Escherichia coli and other Enterobacteriaceae

Humans and animals are ubiquitously colonized by Enterobacteriaceae, a bacterial family that contains both commensals and clinically significant pathogens. Here, we report Enterobacteriaceae megaplasmids of up to 1.58 Mbp in length in infant and adult guts, and other microbiomes. Of 19 complete plasmid genomes, one was reconstructed from an E. coli isolate; others were linked to species of Citrobacter and Enterobacter via analysis of genome modification patterns. The detection of related plasmids in different Enterobacteriaceae, conjugation machinery, and more diverse modified motifs in certain plasmids compared to hosts suggests that these elements are self-transmissible, with a broad host range. The plasmids encode multi-drug efflux systems and potential secreted effectors. Up to 208 tRNAs are encoded and include sequence variants that may counter tRNA-centric defense mechanisms. Overall, the vast megaplasmid coding capacity may broaden host range, increase competitiveness, control invasion by other elements, and counter programmed cell death.

microbiology↗

bioRxiv2025

Ciprofloxacin resistance in Klebsiella pneumoniae: phenotype prediction from genotype and global distribution of resistance determinants

BACKGROUNDCiprofloxacin resistant Klebsiella pneumoniae is common or emerging in many geographies, and knowledge of local resistance rates is important for empirical therapy. Whilst there are known K. pneumoniae ciprofloxacin resistance determinants, there is a lack of systematic data on the effect of determinants, alone and in combination, and there are no publicly accessible tools for predicting resistance from whole genome sequence data. METHODSThe KlebNET-GSP AMR Genotype-Phenotype Group aggregated a matched genotype-phenotype dataset of n=12,167 K. pneumoniae species complex (KpSC) isolates from 27 countries between 2001-2021. We developed a rules-based classifier to predict ciprofloxacin resistance by categorizing the number of quinolone resistance determining regions mutations in gyrA and parC, the number of plasmid-mediated quinolone resistance genes, and the presence/absence of aac(6')-Ib-cr (which can acetylate ciprofloxacin). Predictive performance was assessed using the discovery dataset, for which we re-phenotyped discrepant isolates; and validated using externally contributed datasets (n=7,030 KpSC isolates). RESULTSThe rules-based classifier predicted R vs S/I with categorical agreement, sensitivity, and specificity >96%, and major/very major error rates <4%. Performance was similar across diverse KpSC sources (human, animal, other), species, and intra-species lineages. External validation of the classifier yielded overall 93.12% categorical agreement [95% confidence interval (CI), 92.50-93.74%], 8.65% major errors [95% CI, 7.34-9.97%], and 6.20% very major errors [95% CI, 5.51-6.90%]. We implemented the classifier in Kleborate, a command-line tool that is integrated into the Pathogenwatch web platform. Using this to assess the global distribution of ciprofloxacin resistance determinants in KpSC genomes available in Pathogenwatch (n=31,319, from 109 countries between years 2000-2023), we observed a significant positive association between national quinolone consumption rates and predicted ciprofloxacin resistance (R2=0.20, p=0.004). CONCLUSIONS Ciprofloxacin resistance phenotypes can be reasonably predicted from genotypes, which is sufficient for informing surveillance. However, unexplained resistance remains and accuracy is insufficient for clinical applications. We demonstrate the value of aggregating genotype-phenotype data to explore resistance mechanisms and develop predictors, but highlight complexities in combining phenotype data from different assays and standards.

genomics↗

bioRxiv2025

Machine learning based lineage prediction from AMR phenotypes for Escherichia coli ST131 clade C surveillance across infection types

2.Rising antimicrobial resistance (AMR) in Escherichia coli bloodstream infections (BSIs) in high-income settings has typically been dominated by one clone, the sequence type (ST) 131. More specifically, ST131 clade C (ST131-C) is associated with fluoroquinolone resistance and extended-spectrum {beta}-lactamases (ESBLs). Even though urinary tract infections (UTIs) are a known common precursor to BSIs, there is currently limited knowledge on the longitudinal prevalence of ST131-C in UTIs and, therefore, the temporal link between the two infection types. Leveraging available genomic and antimicrobial susceptibility test (AST) data for ciprofloxacin, gentamicin, and ceftazidime in 2790 E. coli BSI isolates, we trained random forest and Extreme Gradient Boosting (XGBoost) classifiers to predict if an E. coli isolate belongs to ST131-C using only AST data. These models were used to predict the yearly prevalence of ST131-C in 22942 UTI and 24866 BSI isolates from Norway. The XGBoost classifier achieved a prediction F1-score of over 70% on a highly unbalanced dataset where only 4.3% of the genomic BSI isolates belonged to ST131-C. The predicted prevalence of ST131-C in UTIs exhibited a similar annual trend to that of BSIs, with a stable infection burden for eight years after its rapid expansion, confirming that the persistence of ST131-C in BSIs is largely driven by ST131-C UTIs. However, a higher prevalence of ST131-C in BSIs ([~]7%) compared to UTIs ([~]4%) suggests a subsequent enrichment of ST131-C. Our study highlights how existing epidemiological knowledge can be supplemented by utilising extensive data from AMR surveillance efforts without genomic markers. 3. Impact statementThis study proposes a potential analysis method that leverages AST data, which is already regularly collected for AMR surveillance purposes. Using such data to approximate the population-wide prevalence of MDR clones, such as ST131-C, could allow for larger-scale, retrospective studies of its prevalence in a population than genomic-based methods at a significantly lower cost. Such a method could supplement existing knowledge and epidemiology study practices. We use the proposed method to find relationships between the prevalence of the important MDR E. coli clone ST131-C in UTIs and BSIs in Norway. These results suggest that monitoring and reducing MDR in UTIs could reduce the burden of this invasive clone in hard-to-treat BSIs. 4. Data summaryAll AST data, clonal information for isolates with genomic data, and code used in this study can be found in the following repository: https://github.com/theodorross/EColi-UTI-Predictions. Only the clone and published metadata information is shown for the UTI data shared by Handal, Kaspersen et al.

bioinformatics↗

bioRxiv2025

High frequency body site translocation of nosocomial Pseudomonas aeruginosa

Pseudomonas aeruginosa is an important nosocomial pathogen which can cause serious infections across diverse anatomic locations. Infections can spread within an individual to different body sites, but the rate and directionality of this process is unknown. We explored within-host diversity as well as the body site translocation dynamics using de-convoluted metagenomic P. aeruginosa reads from 256 hospital patients sampled at both respiratory and gut sites. Of the 84 patients where P. aeruginosa genomes could be recovered, there were 27 cases where the same P. aeruginosa clone was detected across multiple body sites. Using a simulation approach, we found that the majority of body site sharing was likely due to within-patient translocation of clones rather than independent acquisition from the hospital environment. Using ancestral reconstruction, we predicted that most clones likely occupied a respiratory niche, and that the probable direction of clone transmission was lung-to-gut. We detected within-patient variation and noted a strong enrichment of mutations in genes associated with antimicrobial resistance, irrespective of sample type. There was significantly more translocation than has been previously reported and highlight that lower respiratory tract infections can result in persistent gut colonisation of P. aeruginosa, a major risk factor for sepsis in vulnerable hosts.

microbiology↗

bioRxiv2024

Genomic characterization of a persistent, azole-resistant C. parapsilosis strain responsible for a hospital outbreak during the first COVID-19 wave

Yeasts belonging to the Candida genus typically reside on the mucosal surface and within the respiratory and gastrointestinal tract as commensals. Under conditions of host vulnerability, they can act as opportunistic pathogens, leading to various forms of candidiasis, including candidemia. Such infections can be particularly problematic when caused by isolates that exhibit resistance to antifungal drugs, which is becoming more prevalent in many regions. One hundred and seven samples of Candida spp. were isolated from patients with candidemia in the hospital San Matteo in Pavia (Italy) over a period of 6 years, from 2015 to the first COVID wave in spring 2020. In order to understand the epidemiology of Candida infections in this hospital setting, the isolates were whole-genome sequenced which identified most as C. parapsilosis and C. albicans. Comparative genomics revealed that isolates of C. albicans were genomically diverse, indicating repeated introductions in the hospital from the community. C. parapsilosis isolates comprised two groups of highly similar isolates representing strains capable of long-term persistence in the hospital. All isolates of the main persistent group were resistant to fluconazole and presented variable levels of resistance to voriconazole and itraconazole, resulting from the Y132F substitution in erg11 and the N455D substitution in upc2. Interestingly, with the exception of the single isolate susceptible to both voriconazole and itraconazole, all the 61 isolates presented one unreported missense mutation in mrr1 (S1907C).

genomics↗

bioRxiv2024

The major pathogen Haemophilus influenzae experiences pervasive recombination and purifying selection at local and global scales

Haemophilus influenzae is a major opportunistic human pathogen which causes both non-invasive and invasive disease. The H. influenzae type b (Hib) vaccine has led to a significant reduction of invasive Hib disease, but offers no protection against colonisation or disease by unencapsulated non-typeables (NT) or non-b serotypes, and H. influenzae remains a public health burden worldwide, with increasing reports of multi-drug resistance (MDR). Despite this, there is no comprehensive understanding of the species global population structure. To advance understanding about the evolution and epidemiology of the species, we whole-genome sequenced 4,475 isolates of H. influenzae from an unvaccinated paediatric carriage and pneumonia cohort from northwestern Thailand. Despite no Hib immunisation, serotype b was uncommonly found (5.7%), while 91.7% of isolates were NT. We identified a large number of nearly pan-resistant lineages that were mostly NT, and discovered that no lineages were enriched among disease samples, suggesting the ability to cause invasive disease is not restricted to any subpopulation of the species. Extensive population genetic analyses of our data combined with a worldwide collection of 5,976 published genomes revealed a highly admixed population structure, low core genome nucleotide diversity, and evidence of pervasive negative selection. The combined data confirm that MDR lineages are not confined to our cohort, and their establishment globally is an urgent concern.

microbiology↗

bioRxiv2024

Enhanced metagenomics-enabled transmission inference with TRACS

Coexisting strains of the same species within the human microbiota pose a substantial challenge to inferring the host-to-host transmission of both pathogenic and commensal microbes. Here, we present TRACS, a highly accurate algorithm for estimating genetic distances between strains at the level of individual SNPs, which is robust to intra-species diversity within the host. Analysis of well-characterised Faecal Microbiota Transplantation datasets, along with extensive simulations, demonstrates that TRACS substantially outperforms existing strain aware transmission inference methods. We use TRACS to infer transmission networks in patients colonised with multiple strains, including SARS-CoV-2 amplicon sequencing data from UK hospitals, deep population sequencing data of Streptococcus pneumoniae and single-cell genome sequencing data from malaria patients infected with Plasmodium falciparum. Applying TRACS to gut metagenomic samples from a large cohort of 176 mothers and 1,288 infants born in UK hospitals revealed species-specific transmission rates between mothers and their infants. Notably, TRACS identified increased persistence of Bifidobacterium breve in infants, a finding missed by previous analyses due to the presence of multiple strains.

bioinformatics↗

bioRxiv2024

Basic reproduction number for pandemic Escherichia coli clones is comparable to typical pandemic viruses

Extra-intestinal pathogenic Escherichia coli (ExPEC) ubiquitously colonize the human gut and represent clinically the most significant bacterial species causing urinary tract infections and bacteremia in addition to contributing to meningitis in neonates. During the last two decades, new E. coli multi-drug resistant clones such as ST131, particularly its clades C1 and C2, have spread globally, as has their generally less resistant sister clade ST131-A. Phylodynamic coalescent modeling has indicated exponential growth in the populations corresponding to these clades during the early 2000s. However, it remains unknown how their transmission dynamics compare to viral epidemics and pandemics in terms of key epidemiological quantities such as the basic reproduction number (R0). Estimation of R0 for opportunistic pathogenic bacteria poses a difficult challenge compared to viruses causing acute infections, since data on E. coli infections accumulate with a much longer delay, even in the most advanced public health reporting systems. Here, we developed a compartmental model for asymptomatic gut colonization and onward transmission coupled with a stochastic epidemiological observation model for bacteremia and fitted the model to annual Norwegian national E. coli disease surveillance and bacterial population genomics data. Approximate Bayesian Computation leveraged by the ELFI software package was used to infer R0 for the pandemic ST131 clades. The resulting R0 estimates for ST131-A, ST131-C1 and ST131-C2 were 1.47 (1.31-1.60), 1.18 (1.12-1.20) and 1.13 (1.08-1.20), respectively, indicating that the ST131-A transmission potential can be even comparable to pandemic influenza viruses, such as H1N1. The significantly lower transmissibility of ST131-C1 and ST131-C2 suggests that their global dissemination has been aided by antibiotic selection pressure and that they may be more effectively transmitted through healthcare facilities instead of primarily community-driven transmission. In summary our results provide a fundamental advance in understanding the relative transmissibility of these opportunistic pathogens and that it can vary markedly even between very closely related E. coli.

microbiology↗

bioRxiv2024

Diversity, functional classification and genotyping of SHV β-lactamases in Klebsiella pneumoniae

Interpreting phenotypes of blaSHV alleles in Klebsiella pneumoniae genomes is complex. While all strains are expected to carry a chromosomal copy conferring resistance to ampicillin, they may also carry mutations in chromosomal blaSHV alleles or additional plasmid-borne blaSHV alleles that have extended-spectrum {beta}-lactamase (ESBL) activity and/or {beta}-lactamase inhibitor (BLI) resistance activity. In addition, the role of individual mutations/amino acid changes is not completely documented or understood. This has led to confusion in the literature and in antimicrobial resistance (AMR) gene databases (e.g., NCBIs Reference Gene Catalog and the {beta}-lactamase database (BLDB)) over the specific functionality of individual SHV protein variants. Therefore, identification of ESBL-producing strains from K. pneumoniae genome data is complicated. Here, we reviewed the experimental evidence for the expansion of SHV enzyme function associated with specific amino-acid substitutions. We then systematically assigned SHV alleles to functional classes (wildtype, ESBL, BLI-resistant) based on the presence of these mutations. This resulted in the re-classification of 37 SHV alleles compared with current assignments in NCBIs Reference Gene Catalog and/or BLDB (21 to wildtype, 12 to ESBL, 4 to BLI-resistant). Phylogenetic and comparative genomic analyses support that; i) SHV-1 (encoded by blaSHV-1) is the ancestral chromosomal variant; ii) ESBL and BLI-resistant variants have evolved multiple times through parallel substitution mutations; iii) ESBL variants are mostly mobilised to plasmids; iv) BLI-resistant variants mostly result from mutations in chromosomal blaSHV. We used matched genome-phenotype data from the KlebNET-GSP Genotype-Phenotype Group to identify 3,999 K. pneumoniae isolates carrying one or more blaSHV alleles but no other acquired {beta}-lactamases, with which we assessed genotype-phenotype relationships for blaSHV. This collection includes human, animal, and environmental isolates collected between 2001 to 2021 from 24 countries across six continents. Our analysis supports that mutations at Ambler sites 238 and 179 confer ESBL activity, while most omega-loop substitutions do not. Our data also provide direct support for wildtype assignment of 67 protein variants, including eight that were noted in public databases as ESBL. We reclassified these eight variants as wildtype, because they lack ESBL-associated mutations, and our phenotype data support susceptibility to 3GCs (SHV-27, SHV-38, SHV-40, SHV-41, SHV-42, SHV-65, SHV-164, SHV-187). The approach and results outlined here have been implemented in Kleborate v2.4.1 (a software tool for genotyping K. pneumoniae from genome assemblies), whereby known and novel blaSHV alleles are classified based on causative mutations. Kleborate v2.4.1 was also updated to include ten novel protein variants from the KlebNET-GSP dataset and all alleles in public databases as of November 2023. This study demonstrates the power of sharing AMR phenotypes alongside genome data to improve understanding of resistance mechanisms. Impact statementSince every K. pneumoniae genome has an intrinsic SHV {beta}-lactamase and may also carry additional mobile forms, the correct interpretation of blaSHV genes detected in genome data can be challenging and can lead to K. pneumoniae being misclassified as ESBL-producing. Here, we use matched K. pneumoniae genome and drug susceptibility data contributed from dozens of studies, together with systematic literature review of experimental evidence, to improve our understanding of blaSHV allele variation and mapping of genotype to phenotype. This study shows the value of coordinated data sharing, in this case via the KlebNET-GSP Genotype-Phenotype Group, to improve our understanding of the evolutionary history and functionality of blaSHV genes. The results are captured in an open-source AMR dictionary utilised by the Kleborate genotyping tool, that could easily be incorporated into or used to update other tools and AMR gene databases. This work is part of the wider efforts of the KlebNET-GSP group to develop and support a unified platform tailored for the analysis and interpretation of K. pneumoniae genomes by a wide range of stakeholders. Data summaryBlaSHV allele sequences and class assignments are distributed with Kleborate, v2.4.1, DOI:10.5281/zenodo.10469001. Table S1 provides a summary of blaSHV alleles, including primary accessions, class-modifying mutations, and supporting evidence for class assignments that differ from NCBIs Reference Gene Catalog or BLDB. Whole genome sequence data are publicly available as reads and/or assemblies, individual accessions are given in Table S2; corresponding genotypes and antibiotic susceptibility phenotypes and measurements are available in Tables S3 and S4, respectively.

genomics↗

bioRxiv2024

Convergence and global epidemiology of Klebsiella pneumoniae plasmids harbouring the iuc3 virulence locus.

BackgroundKlebsiella pneumoniae (Kp) is an important pathogen of humans and animals, and recent reports of convergent strains that carry both virulence and antimicrobial resistance genes (ARGs) have raised serious public health concern. The plasmid-borne iuc locus, encoding the siderophore aerobactin, is a key virulence factor in this species. The variant iuc3 is associated with porcine and human clinical isolates and is carried by mostly uncharacterised IncF plasmids. MethodsWe used a combination of short-read and long-read sequencing to characterise IncFIB(K)/IncFII iuc3-carrying plasmids harboured by 79 Kp isolates and one K. oxytoca isolate recovered as part of two large One-Health studies in Italy (SpARK) and Thailand (OH-DART). Adding data from public repositories gave a combined dataset of 517 iuc3 isolates, and the plasmids were analysed using both clustering and phylogenetic methods. FindingsWe note seven large, convergent, plasmids from Thailand that have emerged through the hybridisation of co-circulating plasmids harbouring iuc3 and antimicrobial resistance genes (ARGs) encoding extended-spectrum beta-lactamases (ESBLs). We were also able to identify putative parental plasmids which were mostly associated with two neighbouring meat markets, as were the hybrid plasmids. Clustering and global phylogenetic analyses resolved an iuc3 plasmid sub-group circulating throughout Asia, with occasional examples in Europe and elsewhere. This variant carries multiple ARGs and is commonly harboured by clinical isolates, thus warranting targeted plasmid surveillance. InterpretationOur study reveals that plasmid hybridisation leading to the convergence of resistance and virulence traits may be very common, even in non-clinical ( One-Health) settings. Population-scale plasmid genomics makes it possible to identify putative parental plasmids, which will help to identify plasmid types that are most likely to hybridise, and what the selective consequences may be for the plasmid and host. A distinct iuc3 plasmid sub-variant is associated with clinical isolates in Asia which requires close monitoring. Research In ContextMultiple reports of convergent clones of Klebsiella pneumoniae that combine both hypervirulence and multidrug resistance (MDR-hvKp) have been published recently; a PubMed search in November 2023 using the key words convergence Klebsiella pneumoniae returned 143 papers, 99 of which were published from 2020 onwards. Our study demonstrates that the hybridisation of plasmids carrying AMR and virulence genes is a frequent, ongoing, process in natural populations. The subsequent transfer of plasmids conferring both traits is thus likely to be a key driver behind the spread of convergent strains. Our study also provides an exemplar of how hybrid assemblies can facilitate large-scale global genomic plasmid epidemiology. Evidence before the studyAlthough multiple recent reports highlight the emergence and spread of convergent Kp strains, the confluence of resistance and virulence genes within the same plasmid has not been studied at a population level, and putative parental plasmids are rarely identified. Moreover, there have been few high-resolution genomic epidemiology studies on closely related plasmids using both long and short-read data on a global scale. Added valueWe more than double the number of complete sequences available for plasmids harbouring iuc3 from 58 to 139 and provide evidence on the host lineages most likely to harbour these plasmids (e.g., ST35), and epidemiological source (e.g., pig, wild animal, human). Our comparative analysis of phylogenetic and clustering approaches will help to inform future plasmid epidemiological studies. ImplicationsThe hybridisation of plasmids harbouring virulence and resistance genes occurs frequently in natural populations, even within One-Health settings. However, the selective drivers (if any) and evolutionary consequences of this phenomenon are unclear. There is clear utility in generating closed plasmid genomes on a population scale, and targeted plasmid surveillance on a clinical sub-variant of iuc3 plasmids is warranted.

genomics↗

bioRxiv2023

Genomic tailoring of autogenous poultry vaccines to reduce Campylobacter from farm to fork

Campylobacter is a leading cause of food-borne gastroenteritis worldwide, linked to the consumption of contaminated poultry meat. Targeting this pathogen at source, vaccines for poultry can provide short term caecal reductions in Campylobacter numbers in the chicken intestine. However, this approach is unlikely to reduce Campylobacter in the food chain or human incidence. This is likely as vaccines typically target only a subset of the high strain diversity circulating among chicken flocks and rapid evolution diminishes vaccine efficacy over time. To address this, we used a genomic approach to develop a whole-cell autogenous vaccine targeting isolates harbouring genes linked to survival outside of the host. We hyper-immunised a whole major UK breeder farm to passively target offspring colonisation using maternally-derived antibody. Monitoring progeny, broiler flocks revealed a near-complete shift in the post-vaccination Campylobacter population with a [~]50% reduction in isolates harbouring extra-intestinal survival genes and a significant reduction of Campylobacter cells surviving on the surface of meat. Based on these findings, we developed a logistic regression model that predicted that vaccine efficacy could be extended to target 46% of a population of clinically relevant strains. Immuno-manipulation of poultry microbiomes towards less harmful commensal isolates by competitive exclusion, has major potential for reducing pathogens in the food production chain.

microbiology↗

bioRxiv2023

Escherichia coli plasmidome maps the game of clones

Escherichia coli is the most widely studied microbe in history, but its extrachromosomal elements known as plasmids remain poorly delineated. Here we used long-read technology to high-resolution sequence the entire plasmidome and the corresponding host chromosomes from an unbiased longitudinal survey covering two decades and over 2,000 E. coli isolates. We find that some plasmids have persisted in lineages even for centuries, demonstrating strong plasmid-lineage associations. Our analysis provides a detailed map of recent vertical and horizontal evolutionary events involving plasmids with key antibiotic resistance, competition and virulence determinants. We present genomic evidence of both chromosomal and plasmid-driven success strategies that represent convergent phenotypic evolution in distant lineages, and use in vitro experiments to verify the importance of bacteriocin-producing plasmids for clone success. Our study has general implications for understanding plasmid biology and bacterial evolutionary strategies.

microbiology↗

bioRxiv2023

PAN-GWES: Pangenome-spanning epistasis and co-selection analysis via de Bruijn graphs

Studies of bacterial adaptation and evolution are hampered by the difficulty of measuring traits such as virulence, drug resistance and transmissibility in large populations. In contrast, it is now feasible to obtain high-quality complete assemblies of many bacterial genomes thanks to scalable and affordable long-read sequencing technology. To exploit this opportunity we introduce a phenotype- and alignment-free method for discovering co-selected and epistatically interacting genomic variation from genome assemblies covering both core and accessory parts of genomes. Our approach uses a compact coloured de Bruijn graph to approximate the intra-genome distances between pairs of loci for a collection of bacterial genomes to account for the impacts of linkage disequilibrium (LD). We demonstrate the versatility of our approach to efficiently identify associations between loci linked with drug resistance and adaptation to the hospital niche in the major human bacterial pathogens Streptococcus pneumoniae and Enterococcus faecalis.

bioinformatics↗