Genome Single Nucleotide Polymorphism Research Articles

Decoding the anomalies: a genome-based analysis of Bacillus cereus group strains closely related to Bacillus anthracis

IntroductionThe Bacillus cereus group encompasses a complex group of closely related pathogenic and non-pathogenic bacterial species. Key members include B. anthracis, B. cereus, and B. thuringiensis organisms that, despite genetic proximity, diverge significantly in morphology and pathogenic potential. Taxonomic challenges persist due to inconsistent classification methods, particularly for B. cereus isolates that resemble B. anthracis in genetic clustering.MethodsThis study investigated B. cereus group isolates from blood smears of animal carcasses in Kruger National Park, uncovering an unusual isolate with B. cereus features based on classical microbiological tests yet B. anthracis-like genomic similarities with an Average Nucleotide Identity (ANI) of ≥95%. Using comparative genomics, pan-genomics and whole genome Single Nucleotide Polymorphism (wgSNP) analysis, a total of 103 B. cereus group genomes were analyzed, including nine newly sequenced isolates from South Africa and a collection of isolates that showed some classification discrepancies, thus classified as “anomalous.”Results and discussionOf the 36 strains identified as B. anthracis in GenBank, 26 clustered phylogenetically with the four confirmed B. anthracis isolates from South Africa and shared 99% ANI. Isolates with less than 99% ANI alignment to B. anthracis exhibited characteristics consistent with B. cereus and/or B. thuringiensis, possessing diverse genetic profiles, insertion elements, resistance genes, and virulence genes features, contrasting with the genetic uniformity of typical B. anthracis. The findings underscore a recurrent acquisition of mobile genetic elements within B. cereus and B. thuringiensis, a process infrequent in B. anthracis.ConclusionThis study highlights the pressing need for standardized taxonomic criteria in B. cereus group classification, especially as anomalous isolates emerge. This study supports the existing nomenclature framework which offers an effective solution for classifying species into genomospecies groups. We recommend isolates with ANI ≥99% to standard reference B. anthracis be designated as typical B. anthracis in GenBank to maintain taxonomic clarity and precision.

Emergence of Multidrug-Resistant NDM-5-Producing ST307 Klebsiella pneumoniae in Uruguay, 2023.

Carbapenem and colistin-resistant Klebsiella pneumoniae pose a significant threat to public health, particularly in intensive care units, due to high morbidity and mortality rates. This study aimed to analyze five NDM carbapenemase-producing multidrug-resistant K. pneumoniae isolates from different hospitals. Antimicrobial susceptibility testing, hypermucoviscosity analysis, biofilm production assessment, MLST, PCR, and whole-genome sequencing were conducted. All isolates harbored NDM-5 metallo-β-lactamase, belonging to MLST 307, were biofilm producers and exhibited a stop codon (Q30) along MgrB. Genomic analysis revealed multiple-replicon plasmids carrying resistance genes, notably blaNDM-5, blaCTX-M-15, rmtB, and qnrB1, with complex genetic structures encoding several mobile genetic elements, including the Tn3 family and IS26. All isolates harbored wzi173 (capsule-locus KL102), iutA (a siderophore-associated gene), and the type 3 fimbriae mrkABCDFHIJ operon. The core genome single nucleotide polymorphisms (SNPs) analysis suggests the circulation of two strains of ST307 clone (SNPs range differences 4-77). These findings highlight the potential plasticity of the high-risk ST307 clone and the urgent need for surveillance and intervention strategies to combat antimicrobial resistance. To our knowledge, this is the first report of K. pneumoniae ST307 carrying blaNDM-5 and the first description of ST307 in Uruguay. The presence of blaNDM-5 and pan-aminoglycoside resistance rmtB genes are identified for the first time in Uruguay.

Molecular Characterization of Whole Genome Sequencing of Salmonella spp. in Shapingba District, Chongqing, China, 2016-2023.

In recent years, Salmonella infection is a major global public health concern, particularly in food safety. This study analyzed the genomes of 102 Salmonella strains isolated between 2016 and 2023 from food, foodborne disease patients, and food poisoning incidents, focusing on their molecular characteristics, antibiotic resistance genes (ARGs), and virulence genes. S. enterica serovar Enteritidis (37.3%) and S. enterica serovar Typhimurium (21.6%, including its monophasic variant 1,4,[5],12:i:-) were the main strains among 22 serotypes. Multilocus sequence typing revealed 23 sequence types (STs), with ST11, ST19, and ST34 as the most prevalent. All strains carried at least 24 ARGs. Detection rates for aac(6')-Iy, blaTEM-1, and sul2 ranged from 44.1% to 63.7%, mainly in S. Enteritidis and S. Typhimurium. Rates for qnrS1, sul1, and aadA were 12.8% to 16.7%, while mcr-1 appeared in one ST34 S. Typhimurium strain. All strains contained at least 98 virulence genes. The genes pefABCD, mig-5, and spvBCD were in 48.0% of strains, while rck was found in 36.3%, mainly linked to S. Enteritidis and S. Typhimurium. The tssM gene was found in 37.3% of the strains, exclusively in S. Enteritidis. Core genome single nucleotide polymorphisms (cgSNPs) analysis grouped the strains into nine clusters, with 75.5% belonging to three major groups. Food poisoning event 1 was correlated with cluster 3, while events 2 and 3 were linked to cluster 1. Across events, SNP differences among strains were ≤6. Strains with SNP differences ≤10 were also found in other clusters. This method is promising for tracking sporadic cases and identifying potential foodborne safety incidents. Salmonella strains in the region exhibit substantial genetic diversity, demonstrating the efficacy of cgSNPs analysis for source tracking. Ongoing surveillance is essential given the prevalence of ARGs and virulence genes. This study provides a data foundation for local Salmonella epidemiology.

Genome‐wide SNP Detection in the Horse Mackerel (Trachurus trachurus, T. mediterraneus, T. picturatus) of Turkish Waters Using Illumina and Nanopore Sequencing

The horse mackerel, distributed in the Pacific, Atlantic, Indian Ocean, and Mediterranean waters, is economically valuable due to its high nutritional content and fishing capacity. Global market demand for this species has been increasing over the years. This rising demand causes fishing pressure, endangering the species' sustainability. Especially the horse mackerel species (Trachurus trachurus, Trachurus mediterraneus and Trachurus picturatus), which are found as mixed stocks in Turkish waters and under fishing pressure, should be considered as different stocks and evaluated with distinct management strategies. However, due to the difficulty of distinguishing these three Trachurus species based on morphological characteristics, conflicting reports have emerged, particularly regarding their distribution areas, and species identification is not always precise. In our study, we sequenced the genomes of these species using next-generation sequencing (Illumina) and third-generation sequencing technologies (Oxford Nanopore), and identified SNP (single nucleotide polymorphism) differences to distinguish the species. We identified a total of 2311386 SNPs in the Trachurus trachurus genome, of which 1313612 were transitions and 997774 were transversions. 5099766 SNPs in the Trachurus picturatus genome, of which 2927066 transitions and 2172700 transversions. Similarly, 5007863 SNPs in the Trachurus mediterraneus genome, of which 2940560 transitions and 2067303 transversions. These SNP datasets provide a robust genetic marker toolkit for accurately distinguishing Trachurus species and evaluating genetic diversity within and between populations. The findings of this study contribute essential genetic resources for defining the stock structures of these species, informing sustainable fisheries management strategies, and guiding conservation initiatives. By addressing the challenges of species identification and genetic diversity assessment, this research lays the groundwork for mitigating the impact of overfishing and ensuring the sustainability of horse mackerel populations in Turkish and global waters.

Genotypic diversity and biological characterization of Alphabaculovirus mabrassicae isolates from a wide host range.

Baculoviruses are ideal biological insecticides, providing long-lasting pest control and environmental benefits. Alphabaculovirus mabrassicae stains, with their broad host range, have been effective in agricultural pest management. Various A. mabrassicae isolates (MbMNPV-CHb1/CTa/K1/QD, MyseMNPV-Hb, HearMNPV and MacoNPV-B) have been identified in different hosts. Identifying more effective A. mabrassicae strains with detailed genetic information is crucial for commercial use. Laboratory bioassays showed that the median lethal concentration (LC50) of MyseMNPV-Hb against Mythimna separata was significantly lower than those against Helicoverpa armigera and Spodoptera exigua, but higher than the LC50 of MbMNPV-CHb1, MbMNPV-QD and HearMNPV against H. armigera or S. exigua. Comparative genomic analysis revealed significant differences in genomic composition and single-nucleotide polymorphisms between MyseMNPV-Hb and the other isolates. A piggyBac-like element, likely to have been from Alcis repandata (Lepidoptera: Geometridae), was identified in the genomes of these isolates. Eight genes in the A. mabrassicae genomes were found to be under positive selection. Alphabaculovirus mabrassicae isolates exhibit different infectivity in various pests, indicating the need for selecting appropriate isolates specific target pests. This study elucidates the genetic factors contributing to the differential infectivity of A. mabrassicae isolates and extends knowledge on its population characteristics. © 2025 Society of Chemical Industry.

Phylogeny and virulence determinant detection of Fusobacterium necrophorum strains isolated at the UK Anaerobe Reference Unit between 1982 and 2019.

Explore the presence, or absence, of virulence genes and the phylogeny of a multi-decade UK collection of clinical and reference Fusobacterium necrophorum isolates. Three hundred and eighty-five F. necrophorum strains (1982-2019) were recovered from storage (-80°C). Illumina whole genome sequencing (WGS) was undertaken with 374/385 genomes available for examination after quality checking. Sequences were analysed, using BioNumerics (bioMérieux; v 8.1), for the presence of known virulence genes. Strain phylogeny was investigated using a bespoke Fusobacterium spp. whole genome multi-locus sequence typing (wgMLST) method and single nucleotide polymorphism (SNP) analysis. F. necrophorum ssp. necrophorum and F. necrophorum ssp. funduliforme phylogeny showed clear separation of the two subspecies and clustering of F. necrophorum ssp. funduliforme into three distinct clades. Congruence between SNP and wgMLST analysis was high (99.3%) and indistinguishable clusters were observed with wgMLST (n=3) and SNP (n=1) analysis of isolates derived from different students attending the same education setting. There was no grouping of strains by disease state or decade of isolation. No association was demonstrated with specific virulence gene detection although conspicuous virulence gene patterns were seen amongst the different subspecies and clades. There was no evidence that the pathogenesis of F. necrophorum infection was associated with the presence of the virulence determinants investigated. Host-pathogen interactions should, therefore, be a focus of future research. Person-to-person transmission is a feature of this important pathogen.

Strain phylogroup and environmental constraints shape Escherichia coli dynamics and diversity over a 20-year human gut time series.

Escherichia coli is an increasingly antibiotic-resistant opportunistic pathogen. Few data are available on its ecological and evolutionary dynamics in its primary commensal niche, the vertebrate gut. Using Illumina and/or Nanopore technologies, we sequenced whole genomes of 210 E. coli isolates from 22 stools sampled during a 20-year period from a healthy man (ED) living in Paris, France. All phylogroups, except C, were represented, with a predominance of B2 (34.3%), followed by A and F (19% each) phylogroups. Thirty-five clones were identified based on their haplogroup and pairwise genomic single nucleotide polymorphism distance and classified in three phenotypes according to their abundance and residence time: 25 sub-dominant/transient (52 isolates), five dominant/transient (48 isolates) and five dominant/resident (110 isolates). Four over five dominant/resident clones belonged to B2 and closely related F phylogroups, whereas sub-dominant/transient clones belonged mainly to B1, A and D phylogroups. The long residence times of B2 clones seemed to be counterbalanced by lower colonization abilities. Clones with larger within-host frequency persisted for longer. By comparing ED strain genomes to a collection of commensal E. coli genomes from 359 French individuals, we identified ED-specific genomic properties including an enrichment in genes involved in a metabolic pathway (mhp cluster) and the presence of a very rare antiviral defense island. The E. coli colonization within the gut microbiota was shaped by both the intrinsic properties of the strain lineages, in particular longer residence of phylogroup B2, and the environmental constraints such as diet or phages.

Genetic Concordance of Staphylococcus aureus From Oropharyngeal and Sputum Cultures in People With Cystic Fibrosis.

People with cystic fibrosis (CF) may not expectorate sputum at young ages or after they receive CFTR modulators. While oropharyngeal swabs are commonly used to test for lower airway pathogens, it is unknown whether Staphylococcus aureus from the oropharynx matches the strain(s) infecting the lungs. Our goal was to determine whether oropharyngeal and sputum isolates of S. aureus are genetically distinct in a cohort of patients with CF. We obtained historical S. aureus isolates from patients who intermittently expectorated sputum in 2018, and we prospectively cultured S. aureus from oropharyngeal swabs and sputum from subjects with CF between August 2020 and February 2022. We performed short-read whole genome sequencing, determined sequence type, and performed phylogenetic analysis using S. aureus core genome single nucleotide polymorphisms (SNPs). We assigned isolates from a patient to the same strain if they had the same sequence type and differed by ≤ 60 SNPs or the isolates were not disturbed by clade breaker analysis. 36 subjects had S. aureus in ≥ 1 oropharyngeal swab and ≥ 1 sputum in 2018. In the prospective collection, 31 subjects had synchronous oropharyngeal swab and sputum collections. Although polyclonal infections were detected, sputum and oropharyngeal isolates of S. aureus typically matched the same strain within study subjects, both over the span of 2018 (31/36 patients) and when collected simultaneously from 2020 to 2022 (29/31 patients). In patients with CF who intermittently produce sputum, oropharyngeal swabs identify S. aureus with genetic and phenotypic similarity to those cultured from sputum.

Rapid whole genome characterization of antimicrobial-resistant pathogens using long-read sequencing to identify potential healthcare transmission.

Whole genome sequencing (WGS) can help identify transmission of pathogens causing healthcare-associated infections (HAIs). However, the current gold standard of short-read, Illumina-based WGS is labor and time intensive. Given recent improvements in long-read Oxford Nanopore Technologies (ONT) sequencing, we sought to establish a low resource approach providing accurate WGS-pathogen comparison within a time frame allowing for infection prevention and control (IPC) interventions. WGS was prospectively performed on pathogens at increased risk of potential healthcare transmission using the ONT MinION sequencer with R10.4.1 flow cells and Dorado basecaller. Potential transmission was assessed via Ridom SeqSphere+ for core genome multilocus sequence typing and MINTyper for reference-based core genome single nucleotide polymorphisms using previously published cutoff values. The accuracy of our ONT pipeline was determined relative to Illumina. Over a six-month period, 242 bacterial isolates from 216 patients were sequenced by a single operator. Compared to the Illumina gold standard, our ONT pipeline achieved a mean identity score of Q60 for assembled genomes, even with a coverage rate as low as 40×. The mean time from initiating DNA extraction to complete analysis was 2 days (IQR 2-3.25 days). We identified five potential transmission clusters comprising 21 isolates (8.7% of sequenced strains). Integrating ONT with epidemiological data, >70% (15/21) of putative transmission cluster isolates originated from patients with potential healthcare transmission links. Via a stand-alone ONT pipeline, we detected potentially transmitted HAI pathogens rapidly and accurately, aligning closely with epidemiological data. Our low-resource method has the potential to assist in IPC efforts.

Genomic and Gut Microbiome Evaluations of Growth and Feed Efficiency Traits in Broilers.

In this study, we combined genomic and gut microbiome data to evaluate 13 economically important growth and feed efficiency traits in 407 Dahen broilers, including body weight (BW) at four, six, nine, and ten weeks of age (BW4, BW6, BW9, and BW10), as well as the average daily gain (ADG6, ADG9, and ADG10), feed conversion ratio (FCR6, FCR9, and FCR10), and residual feed intake (RFI6, RFI9, and RFI10) for the three growing ages. The highest ADG and lowest FCR were observed at nine and six weeks of age, respectively. We obtained 47,872 high-quality genomic single-nucleotide polymorphisms (SNPs) by sequencing the genomes and 702 amplicon sequence variants (ASVs) of the gut microbiome by sequencing the 16S rRNA gene, both of which were used for analyses of linear mixed models. The heritability estimates (± standard error, SE) ranged from 0.103 ± 0.072 to 0.156 ± 0.079 for BW, 0.154 ± 0.074 to 0.276 ± 0.079 for the ADG, 0.311 ± 0.076 to 0.454 ± 0.076 for the FCR, and 0.413 ± 0.077 to 0.609 ± 0.076 for the RFI traits. We consistently observed moderate and low negative genetic correlations between the BW traits and the FCR and RFI traits (r = -0.562 to -0.038), whereas strong positive correlations were observed between the FCR and RFI traits (r = 0.564 to 0.979). For the FCR and RFI traits, strong positive correlations were found between the measures at the three ages. In contrast to the genomic contribution, we did not detect a gut microbial contribution to all of these traits, as the estimated microbiabilities did not confidently deviate from zero. We systematically evaluated the contributions of host genetics and gut microbes to several growth and feed efficiency traits in Dahen broilers, and the results show that only the host genetics had significant effects on the phenotypic variations in a flock. The parameters obtained in this study, based on the combined use of genomic and gut microbiota data, may facilitate the implementation of efficient breeding schemes in Dahen broilers.

Phenotypic Plasticity, Multiple Paternity, and Shell Shape Divergence Across Lake‐Stream Habitats in a Freshwater Mussel Brood (Pyganodon grandis)

ABSTRACT Hydrodynamic forces and their absence appear to exert differential selection pressure on aquatic biodiversity in lake and stream habitats, creating a tight fit between organismal phenotypes and their environments. Ecophenotypic variants may be the result of genetic differentiation or phenotypic plasticity, where a genotype can produce multiple phenotypes dependent on the environment. Freshwater mussels possess a wide degree of morphological variation that frequently covaries with the environment, making them a good system to understand the mechanisms of ecophenotypic variation across hydrological conditions. We designed a two‐year experiment where individuals from the same Pyganodon grandis maternal brood (half and full siblings) were reared at a controlled site and four natural sites involving one lake and three streams. At the end of the experiment, shell shape was quantified for recaptured (N = 70), wild (N = 206), and zoo‐reared (N = 305) mussels. The maternal individual and 46 recaptured mussels were sequenced for genomic single nucleotide polymorphisms to test for multiple paternity and its effect on offspring morphology. Analysis of covariance found significant differences in shell shape between rearing sites, particularly between stream and lake habitats, but no shape differences were detected across the three stream sites. At two of the four sites, the shell shape of recaptured individuals was not significantly different than that of wild populations. Genomic sequencing and parentage analysis identified 11–27 different fathers among recaptured individuals. Yet no genetic differences were present between stream and lake habitats, and there was no paternal effect on shell shape. Taken together, phenotypic plasticity, over genetic differentiation, is identified as the primary mechanism of ecophenotypic variation. Plasticity is likely ubiquitous across freshwater mussels and may be a key adaptation given their high variance in habitat use. Multiple paternity may also play a role in the evolution of phenotypic plasticity, allowing more males from greater distances opportunities for fertilization, thus increasing genetic connectivity. Lastly, phenotypic plasticity and multiple paternity are convenient properties for freshwater mussel conservation and propagation. Multiple fathers increase the genetic variation of propagated broods, while plasticity may provide resilience to the release of stocked individuals across environmental heterogeneity.

Extensive cytonuclear discordance revealed by phylogenomic analyses suggests complex evolutionary history in the holly genus Ilex (Aquifoliaceae)

Ilex L., the exclusive genus of Aquifoliaceae, encompasses over 600 dioecious wood species with a highly irregular distribution, predominantly found in South America and Asia. The phylogeny and classification of this genus remain enigmatic due to significant early extinctions, constrained morphological diversity, recent hybridization/introgression, and conflicting signals from previously utilized markers. This study presents phylogenetic reconstructions based on complete chloroplast genome sequences and single nucleotide polymorphisms (SNPs) derived from genome resequencing data. A total of 116 accessions of Ilex, representing approximately 108 taxa, were included as the ingroup, with five accessions of two species serving as outgroups. Analysis of the chloroplast genome and nuclear SNP data individually resulted in two robust phylogenetic trees, revealing substantial discrepancies between the chloroplast genome and nuclear SNP phylogenies at both the species and clade levels. The chloroplast genome sequences successfully resolved relationships within this genus into eight strongly supported major clades, while the nuclear SNPs resolved relationships into seven highly supported major clades. Our nuclear SNP phylogenetic tree, in comparison to the chloroplast genome tree, aligns more closely with the recently updated classification of Ilex in multiple instances. The extensive cytonuclear discordance identified may be attributed to recent hybridization events and incomplete lineage sorting (ILS).

Genomic epidemiological analysis of a single-centre polyclonal outbreak of Serratia marcescens, Belgium, 2022 to 2023.

Serratia marcescens is an opportunistic pathogen with a propensity to cause nosocomial outbreaks, particularly in neonatal intensive care units (NICUs). We present a sustained outbreak spanning over 18 months (1 January 2022-29 August 2023) in a NICU in Antwerp, Belgium, affecting 61 neonates, identified through samples taken for diagnostic purposes and by rectal screening. Ten neonates were infected: five with lower respiratory tract infection, four with conjunctivitis and one fatal case with sepsis. In a logistic regression analysis, nursing in an incubator was significantly associated with acquisition of S. marcescens (odds ratio (OR): 2.99; 95% confidence interval (CI): 1.14-8.25; p < 0.05). Whole genome sequencing-based multilocus sequence typing (wgMLST) and core genome single nucleotide polymorphism (cgSNP) analysis of isolates from clinical (n = 4), screening (n = 52) and environmental samples (n = 8), identified eight clusters and five singletons not associated with the clusters. Although outbreak measures were successful in containing further spread within the ward during sudden surges when > 4 cases per week were identified (peak events), several peaks with different clonal clusters occurred. The emergence of similar outbreaks in Belgian hospitals underscores the need of continuous surveillance and NICU-specific infection prevention and control (IPC) measures.

Genomic Single Nucleotide Polymorphism (SNP) markers and mitochondrial haplotypes illuminate the origins of Crown-of-Thorns Starfish (Acanthaster solaris) outbreaks in the South China Sea

BackgroundOutbreaks of the coral predator Crown-of-Thorns Starfish (CoTS) pose a severe threat to coral reefs in the Indo-Pacific Ocean. In 2018, the South China Sea (SCS) experienced significant CoTS outbreaks, leading to extensive coral mortality across the Xisha, Zhongsha, Dongsha, and Nansha Islands, severely impacting the coral reef ecosystem.ResultsTo explore the origins of these outbreaks, we conducted a comprehensive genomic analysis using data from genomic single nucleotide polymorphism sites (SNPs) and mitochondrial haplotypes. Our analysis reveals that CoTS populations in the SCS, which exhibit moderate genetic diversity and may have undergone positive selection or population expansion. There was limited genetic differentiation among CoTS populations from XS, ZS, and NS groups. Especially between the XS and ZS groups, there was almost no genetic differentiation. The populations from XS, ZS, and NS groups have strong genetic connections with populations in Vietnam and the Philippines. There was high gene flow from Vietnam to the Xisha Islands and from the Philippines to the Nansha Islands, suggesting that the CoTS populations in these regions primarily originate from these neighboring countries.ConclusionThe comprehensive analyses of SNP and mitochondrial genomes have provided valuable insights into the population genetics of CoTS. This research has generated significant genomic resources and facilitated important studies on the genetics of the CoTS species. By identifying potential source populations and understanding the genetic basis of their spread, managers can develop more effective conservation strategies to protect vulnerable coral reef ecosystems in the SCS.

Validation and implementation of whole-genome sequencing-based analytical methods for molecular surveillance and relatedness analysis of Mycobacterium tuberculosis complex isolates at a national reference laboratory

Whole genome sequencing-based methodologies have become extremely relevant for the molecular surveillance of human pathogens and are being increasingly introduced into national reference laboratory services. In this study, we describe the validation and implementation of core-genome Multi-Locus Sequence Typing (cgMLST) and whole genome single-nucleotide polymorphism (wgSNP) analysis at the Irish Mycobacteria Reference Laboratory, as a replacement for Mycobacterial Interspersed Repetitive Unit-Variable Number Tandem Repeat (MIRU-VNTR) typing. Concordance of clustering, discriminatory power, and ease-of-use of both WGS analytical methods were evaluated. Although wgSNP analysis (MTBseq) was the most discriminatory method (p < 0,001), we recommend cgMLST (SeqSphere+), as the first-line approach for molecular typing of Mycobacterium tuberculosis isolates in the context of routine surveillance work due to its ease of use and decreased turnaround time, while reserving wgSNP-analysis for a more in-depth cluster analysis of new isolates that show a distance of ≤12 alleles to any other isolate(s) in the cgMLST database.

Whole-genome sequencing of clinical isolates of Klebsiella michiganensi in China carrying blaIPM-4 and blaNDM-1

AimThis study aimed to analyze the prevalence, phenotypes, and carriage of resistance genes of carbapenem-resistant Klebsiella michiganensis strains isolated in our hospital. Methodology: Four K. michiganensis strains were collected from January 2015 to December 2023. Antimicrobial susceptibility was tested using 21 antibiotics with the BD Phoenix™ M50 System. Whole-genome sequencing of the four strains was performed on an Illumina NovaSeq 6000 platform. Species identification was performed using Kleborate software, sequence type (ST) was performed using MLST, and prediction of antibiotic resistance genes and virulence genes were performed using ABRicate. PlasmidFinder was used to search for plasmids. Whole genome data from 211 strains of carbapenem-resistant K. michiganensis were downloaded from the NCBI database; together with the strains in this study, these data were used to construct an phylogenetic tree using genome single nucleotide polymorphisms (SNP). ResultsAntimicrobial susceptibility showed that K. michiganensis was highly resistant to β-lactams. For the three carbapenems, strain WF0046 was only resistant to ertapenem, while WF0047, WF0052, and WF0053 were resistant to all three carbapenems tested. The four strains of K. michiganensis only showed complete sensitivity to polymyxin E and tigecycline. The four K. michiganensis strains were found to have 43 resistance genes, and all carried carbapenem resistance genes; WF0046 carried the carbapenem resistance gene blaIMP-4, while the other three strains carried blaNDM-1. Among the other resistance genes, β-lactam resistance genes were predicted most (11 types), followed by eight types of aminoglycoside resistance genes. Of the strains characterized in this study, WF0046 belonged to ST158, WF0047 belonged to ST40, WF0052 belonged to ST533, and WF0053 belonged to ST13. These four strains, along with 211 carbapenem-resistant K. michiganensis strains downloaded from NCBI, were divided into five clades; WF0052 belonged to clade A, WF0046 belonged to clade C, and WF0047 and WF0053 both belonged to clade D. The four strains had relatively distant genetic relationships; WF0046, WF0047, and WF0053 were closely related to strains of human origin from other regions of China, while WF0052 was genetically close with a strain of K. michiganensis isolated in the United States. ConclusionThe strains of K. michiganensis from our hospital were resistant to multiple antibiotics, and all strains carried carbapenem resistance genes along with multiple other antibiotic resistance genes. The phylogenetic results showed that these four strains had distant genetic relationships and different ST types, indicating that they came from different sources and the possibility of polyclonal transmission.

Potential use of Fourier-transform infrared spectroscopy as a rapid screening tool for investigating nosocomial outbreaks of ST-80 vancomycin-resistant Enterococcus faecium

Core genome single-nucleotide polymorphism phylogeny was used to characterise a nosocomial outbreak caused by ST-80 Vancomycin-resistant Enterococcus faecium (VREf). It identified 22 of 25 epidemiologically related isolates as belonging to an outbreak cluster. The use of Fourier-transform infrared (FT-IR) spectroscopy with a cluster-defining cut-off of 0.071 resulted in the correct classification of 21 of 22 phylogenetically related isolates in a single cluster. It successfully distinguished three phylogenetically unrelated isolates from the outbreak cluster, along with five ST-80 unrelated control isolates, and five isolates from a previous outbreak in May 2023, yielding only one mischaracterised environmental isolate. These findings support the potential use of FT-IR spectroscopy as a rapid screening tool to assist outbreak investigations. Notably, this study is the first to focus on the performance of FT-IR spectroscopy in the epidemiological analysis of VREf isolates with the same sequence type.

Genetic structure and common ancestry expose the dingo-dog hybrid myth.

The evolutionary history of canids has been shown to be complex, with hybridization and domestication confounding our understanding of speciation among various canid lineages. The dingo is a recent canid lineage that was completely isolated from other canids for over 5000 years on the Australian mainland, but the introduction of domestic dogs in 1788 has placed doubt on its independence, with recent studies highlighting hybridization between dingoes and domestic dogs. Using genomic single nucleotide polymorphism data from 434 Australian canid samples, we explicitly test for introgression between closely related canid groups and dingoes. We found no evidence of introgression between dingoes and domestic dogs and show that previous work has likely mischaracterized shared ancestral genetic variation as evidence for hybridization. Further, New Guinea Singing Dogs are the only canid group that significantly shared genetic variation with dingoes, which fits with our understanding of previous phylogenetic analyses. Despite more recent sympatric distributions with dogs, dingoes have likely maintained their independence since their arrival in Australia, even in areas with high lethal control, indicating that their evolutionary trajectory is currently being conserved. The future conservation of the dingo lineage will require policies that promote coexistence pathways between humans and dingoes that protect rangeland systems and the dingoes' evolutionary future.

Genome-wide species delimitation and quantification of the extent of introgression in eriophyoid mite Epitrimerus sabinae complex (Acariformes: Eriophyoidea)

Species complex hinders the exploration of terrestrial species diversity, particularly in small arthropod lineages that are morphologically indistinguishable from each other. The Epitrimerus sabinae complex in the Eriophyoidea provides a valuable case study in species complex delimitation, as they exhibit limited morphological variations. In this study, we obtained thousands of nuclear genomic single-nucleotide polymorphisms via whole-genome sequencing from 55 E. sabinae complex specimens, covering their potential all known distribution ranges. We implemented a framework to infer cryptic speciation, which involved phylogenetic and genetic clustering to identify potential species, followed by population demographic assessment to confirm lineage independence (and thus species status). Our results demonstrate that the E. sabinae complex comprises ten distinct species. These species range from highly divergent, genetically isolated lineages, to differentiated populations involving gene flow. This gene flow is widespread across species boundaries, indicating potential genetic introgression among them. Additionally, demographic analyses revealed that the ten species have followed unique trajectories in size change during the Quaternary period. Time-calibrated phylogenies further showed that speciation in the E. sabinae complex occurred rapidly, resulting in a rapid radiation during the Neogene period. Collectively, parallelism/convergence and recent divergence involving multiple gene flows may explain the homoplasy of E. sabinae complex. Our results highlight the integrated approach in species complex delimitation.

Comprehensive genome-wide analysis of genetic loci and candidate genes associated with litter traits in purebred Berkshire pigs of Korea.

The objective of this study was to identify genomic regions and candidate genes associated with the total number of piglets born (TNB), number of piglets born alive (NBA), and total number of stillbirths (TNS) in Berkshire pigs. This study used a total of 11,228 records and 2,843 single-nucleotide polymorphism (SNP) data obtained from Illumina porcine 60 K and 80 K chips. The estimated genomic breeding values (GEBVs) and SNP effects were estimated using weighted single-step genomic BLUP (WssGBLUP). The heritabilities of the TNB, NBA, and TNS were determined using single-step genomic best linear unbiased prediction (ssGBLUP). The heritability estimates were 0.13, 0.12, and 0.015 for TNB, NBA, and TNS, respectively. When comparing the accuracy of breeding value estimates, the results using pedigree-based BLUP (PBLUP) were 0.58, 0.60, and 0.31 for TNB, NBA, and TNS, respectively. In contrast, the accuracy increased to 0.67, 0.66, and 0.42 for TNB, NBA, and TNS, respectively, when using WssGBLUP, specifically in the last three iterations. The results of weighted single-step genome-wide association studies (WssGWAS) showed that the highest variance explained for each trait was predominantly located in the Sus scrofa chromosome 5 (SSC5) region. Specifically, the variance exceeded 4% for TNB, 3% for NBA, and 6% for TNS. Within the SSC5 region (12.26 to 12.76 Mb), which exhibited the highest variance for TNB, 20 SNPs were identified, and five candidate genes were identified: TIMP3, SYN3, FBXO7, BPIFC, and RTCB. The identified SNP markers for TNB, NBA, and TNS were expected to provide valuable information for genetic improvement as an understanding of their expression and genetic architecture in Berkshire pigs. With the accumulation of more phenotype and SNP data in the future, it is anticipated that more effective SNP markers will be identified.