Infection, Genetics and Evolution

Group A Streptococcus (GAS), or Streptococcus pyogenes is almost exclusive and greatly adapted human pathogen. It causes a wide array of clinical symptoms, ranging from minor infections of the skin and soft tissues to pharyngitis, meningitis, pneumonia, bacteraemia, cellulitis, puerperal sepsis, and necrotising fasciitis. The risk of S. pyogenes infection is known to be influenced by several host characteristics, including age, underlying diseases like diabetes, varicella, or skin lesions, both chronic and acute, and certain risk behaviours such as use of drugs. Household size and overcrowding are two environmental factors that significantly affect the transmission of S. pyogenes. The majority of cases occur spontaneously in the community, and preventative opportunities are still limited. A large portion of GAS-related mortality is found in low-income areas and communities. Based on aforementioned public health risk, the creation of effective therapeutic vaccines would be an excellent addition to current control measures. The purpose of this work is to address the need for new instruments to aid in the elimination of S. pyogenes infections. The discovery of high antigenic regions in several highly conserved proteins brings us one step closer to developing peptide vaccines capable of influencing the different phases of S. pyogenes infection, providing more effective defence and greater serotype coverage. This study used various techniques of immunoinformatics to design an effective multi-epitope vaccine that produced neutralising antibodies against multiple strains of S. pyogenes.

BackgroundOutbreaks of fluconazole-resistant Candida parapsilosis have recently emerged worldwide. In Spain, this phenomenon has been reported since 2020, mainly involving isolates from different clones harbouring the Y132F mutation at Erg11. MethodsWe analysed the expansion of fluconazole resistant C. parapsilosis strains within the national antifungal resistance surveillance program. Genetic clustering and relationships were assessed using microsatellite typing and whole genome sequencing. FindingsWe identified the expansion of three distinct clones carrying the Y132F mutation. Additionally, there was an increase in strains harbouring the G458S mutation, most of which belonged to a clonal complex, although other less prevalent clones were also detected. G458S isolates showed higher resistance to azoles than Y132F strains, particularly to voriconazole and isavuconazole. This increased resistance was associated with mutations in the Tac1 transcriptional regulator and duplication of a chromosomal region containing Tac1 and Erg11. One G458S isolate without mutation at Tac1 exhibited lower MIC values. Furthermore, two isolates carried the K143R mutation, and a distinct group of resistant strains without detectable ERG11 mutations was also identified. Resistant cases were detected across 31 hospitals in 12 autonomous regions. InterpretationOur findings indicate a concerning nationwide expansion of antifungal-resistant C. parapsilosis in Spain, involving multiple resistance mechanisms and clonal lineages, with implications for antifungal treatment and infection control strategies.

We report the genome sequence of Bacillus paranthracis SCM10-01, isolated from a wild neotropical bird (Synallaxis cabanisi) collected in Peru. The assembly yielded one chromosome, three plasmids, and Bacillus phage SCM10. Genomic screening identified complete hemolysin BL, nonhemolytic enterotoxin operons, and cytotoxin K2, but no anthrax-associated toxin or capsule genes.

Among Madagascar primates, the sportive lemurs (family Lepilemuridae) have seen their species diversity increase from eight in 2005 to 26 in 2009 mostly by applying the phylogenetic species concept to DNA barcode data. Despite the genus being speciose, only one case of sympatry is known from northern Madagascar, where two sportive lemur species described based on low mtDNA divergence, Lepilemur ankaranensis and Lepilemur milanoii, were found to co-occur at the center of their joint distribution range. Here, to clarify the taxonomy of these two species and examine their sympatry, we apply an integrative taxonomic framework to genomic and morphological data from 84 individuals of L. ankaranensis and L. milanoii, encompassing their entire distribution range and the forest of Analafiana, beyond their southernmost limit. Using clustering, multivariate, and isolation by distance analyses, we find no evidence of a sympatric zone and show that despite clear genetic differentiation between regions, the genomic and morphological diversity of the L. ankaranensis, L. milanoii-Analafiana group is clinal and explained by geographic distance. These results clarify that L. milanoii is a junior synonym of L. ankaranensis and that the Analafiana forest population belongs to L. ankaranensis, extending its distribution. It further implies that the 'sympatric' zone, the Andrafiamena forest, hosts conspecific individuals with slightly differentiated mtDNA backgrounds, rather than slightly differentiated sympatric species. Lastly, we re-evaluate the IUCN conservation metrics of L. ankaranensis, which continue to qualify as Endangered (EN) under the B1ab(i-v) criteria.

BackgroundMalaria control is increasingly challenged by the urban-adapted vector Anopheles stephensi, yet molecular and cellular tools for this species remain scarce, restricting functional genomic studies and the development of genetic control strategies. To help address this gap, we established a new embryo-derived Anopheles stephensi cell line. ResultsWe generated and characterised a novel embryo-derived Anopheles stephensi (SDA-500) cell line capable of sustained growth in vitro. Species identity was confirmed by mitochondrial COI barcoding, and karyotypic analysis revealed a diploid chromosome complement with the presence of a Y chromosome, confirming that at least some cells are of male origin. Transfection conditions were optimized, with TransIT-PRO showing higher efficiency than Lipofectamine-based reagents. Using a dual-luciferase reporter assay, of several promoters tested the Anopheles gambiae polyubiquitin promoter exhibited the strongest and most consistent transcriptional activity in SDA-500 cells. ConclusionsThe SDA-500 cell line provides a stable and genetically validated in vitro platform that supports efficient transgene expression. This resource provides a useful system for functional genomics and molecular manipulation in Anopheles stephensi and is expected to facilitate studies of mosquito biology and contribute to the development of novel malaria control strategies.

IntroductionIn sub-Saharan Africa, and more specifically in Cameroon, antimicrobial resistance (AMR) represents a major public health threat. This is underlined by the increasing appearance of multidrug-resistant bacteria. Extended-spectrum {beta}-lactamase producing Escherichia coli (ESBL-Ec), a critical priority bacterium, is increasingly implicated in life-threatening infections in hospital and community settings in Cameroon. Data on the genetic composition of ciprofloxacin-resistant Escherichia coli are limited in Cameroon. This study aimed to investigate the prevalence, genetic diversity, resistance mechanisms in multidrug-resistant Escherichia coli organisms isolated from clinical samples in two hospitals in Yaounde, Cameroon. MethodA cross-sectional study was conducted from February to June 2025 in two healthcare facilities in Yaounde, Cameroon. All clinical samples from in- and out-patients were analysed. After culturing, identification was performed using API20E as per the manufacturers instructions and ESBL production was screened in CHROMagarTM ESBL. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disc diffusion method. Polymerase chain reaction (PCR) was used to detect ESBL and plasmid mediated quinolone resistance (PMQR)genes, as well as mutations in quinolone resistance-determining region (QRDR) (gyrA/parC) Horizontal. plasmid transfer was also investigated. Finally, phylogroup analysis was assessed. ResultThe prevalence of MDR E. coli was 50.7% (n=33/65), all of which (100%) were ESBL producers and 91% were ciprofloxacin-resistant. Highest resistance rates were observed for cefotaxime (100%), ceftriaxone (100%), and ciprofloxacin (91%). The most frequent ESBL genes were blaTEM (36.3%; n=12/33). Among PMQR genes, qnrB was detected in 16.6% (n=5/30) of isolates. Only the ESBL genes were carried by plasmids; the most prevalent plasmid-borne gene was blaTEM (40%), followed by blaCTX-M (26.7%). Mutations within the topoisomerase QRDR (parC gene) were identified in 36.6% (n=11/30) of ciprofloxacin-resistant strains. Phylogroup analysis revealed a predominance of phylogroup A, followed by group B. ConclusionThis study reveals a high prevalence of multidrug-resistance, ESBL (blaTEM dominant) and fluoroquinolone resistance in E. coli in Yaounde, with plasmid dissemination of ESBL genes and chromosomal stabilization of PMQR determinants. The predominance of commensal phylogroups in clinical samples underlines the role of the community reservoir. It is urgent to reinforce " real-time One Health" genomic surveillance in Cameroon.

BackgroundThe ecological dynamics between Trypanosoma parasites and their wild mammalian hosts, such as bats and armadillos, are complex. Recent 18S rRNA metabarcoding studies have reported extraordinary levels of hidden parasite diversity and frequent multi-lineage coinfections within individual wild hosts. However, the boundary between genuine biological coinfection and methodological artifact remains difficult to establish. Based on Gauses principle of competitive exclusion, the mammalian bloodstream represents a highly constrained niche where stable coexistence of identical ecological competitors is theoretically rare. We hypothesize that previously reported hyper-diverse Trypanosoma coinfections are largely bioinformatic artifacts, and that true intra-host dynamics instead favor single-lineage dominance. MethodsTo test this hypothesis, we sequenced samples from 27 wild armadillos (Dasypus novemcinctus) and 26 bats from Ecuador. The 18S rRNA gene was amplified via nested PCR and sequenced using an Oxford Nanopore Technologies MinION platform. We developed a progressively stringent bioinformatics pipeline to evaluate coinfection hypotheses. Raw reads were processed through three alignment scenarios: Lenient, Moderate, and Conservative. These scenarios modulate sequence identity, mapping quality (MAPQ), and coverage thresholds to effectively isolate true biological signals from alignment ambiguity. ResultsUnder lenient alignment parameters, the resulting profiles mirrored previous literature, exhibiting massive apparent intra-host multi-lineage diversity. However, as bioinformatic stringency increased to conservative thresholds ([≥] 98% sequence identity, [≥] 99% coverage, and MAPQ [≥] 30), artifactual pseudo-coinfections collapsed. The highly restricted dataset demonstrated overwhelming single-lineage dominance, validating only three active mixed infections out of the retained samples. Furthermore, our rigorous pipeline isolated rare but genuine biological signals, including the detection of Trypanosoma cruzi marinkellei--historically considered a bat-restricted subgenus--within the terrestrial armadillo cohort. We also confirmed the presence of T. cruzi DTU III (TcIII) in Ecuadorian armadillos, representing a significant biogeographical record for the region. ConclusionsOnce methodological noise is computationally stripped away, active multi-strain Trypanosoma coinfections in the host bloodstream are revealed to be ecologically anomalous. Our findings strongly support the principle of competitive exclusion, suggesting established lineages actively suppress competitors. While Oxford Nanopore sequencing offers necessary resolution for wildlife parasitology, fine-tuning algorithmic parameters is critical to accurately represent host-parasite networks and prevent the artificial inflation of intra-host diversity metrics. Author summaryPrevious studies using DNA metabarcoding have reported that wild mammals, such as bats, frequently harbor complex communities of multiple Trypanosoma parasite lineages simultaneously. However, ecological principles suggest that identical competitors struggle to coexist stably within a constrained environment like the host bloodstream. To investigate whether these reported high coinfection rates reflect true biology or methodological artifacts, we sequenced the 18S rRNA gene of Trypanosoma from 26 bats and 27 armadillos in Ecuador. We processed the sequencing data through computational pipelines with progressively stricter filtering parameters. We observed that under lenient filtering, animals appeared to have highly diverse, mixed infections. Conversely, when strict parameters were applied to remove potential analytical noise, the artificial complexity collapsed, revealing that the vast majority of hosts were dominated by a single parasite lineage. We confirmed only three active mixed infections in our highly restricted dataset. Our findings indicate that active multi-strain Trypanosoma coinfections are rare, aligning with the principle of competitive exclusion. These results highlight the necessity of applying rigorous bioinformatic filters to accurately evaluate host-parasite interactions and avoid overestimating diversity metrics.

Most cockroaches and the termite Mastotermes darwiniensis are associated with Blattabacterium, an ancient obligate endosymbiont that participates in the nitrogen metabolism of its host. Blattabacterium has been vertically transmitted since it was acquired by the common ancestor of cockroaches and termites and was reportedly lost twice, once in the cockroach genus Nocticola and once in all termites except Mastotermes darwiniensis. Here, we acquired cockroach specimens spanning most of the cockroach phylogenetic tree to study Blattabacterium using shotgun sequencing. We found no traces of Blattabacterium in 64 specimens from ten independent lineages of cockroaches across three families: Blattellidae, Pseudophyllodromiidae, and Anaplectidae. The absence of Blattabacterium was confirmed with three PCR amplifications targeting the 16S and 23S ribosomal genes with primers specific to Blattabacterium. Notably, cockroaches lacking Blattabacterium were often infected by Rickettsia and Wolbachia, many of which were related to the mutualistic Wolbachia strain of Cimex lectularius, the common bed bug. These results indicate that cockroaches from Blattellidae, Pseudophyllodromiidae and Anaplectidae have lost their ancestral Blattabacterium endosymbiont at least ten times independently, with many of these losses possibly facilitated and compensated by new associations with mutualistic Wolbachia strains that may help provision the host with B vitamins.

Based on recent epidemiologic studies, Streptococcus agalactiae (Group B Streptococcus; GBS) sequence type (ST) 1010 is an emerging lineage now identified in multiple countries. We report the phylogenetic and genomic characteristics of a set of 55 GBS sequence type (ST) 1010 strains, as well as two newly described single-locus variants of ST1010. A core genome phylogeny suggests that ST1010 is closely related to both ST452 and the hypervirulent clonal complex (CC) 17 GBS lineage. Notably, we demonstrate that genes encoding two virulence determinants previously described as specific to CC17 GBS, the HvgA adhesin and the serine-rich repeat protein Srr2, are both present in ST1010 genomes. Srr2 is shared with members of ST452. High-level gentamicin resistance (HLGR) encoded on an IS256 mobile element, previously described in a small number of ST1010 isolates, is present in a distinct ST1010 subclade encompassing the majority of ST1010 isolates. The relationship between ST452 (serotype IV), ST1010 (serotype IV), and ST17 (serotype III) strains suggests that ST17 may have arisen from a serotype IV ancestor and later acquired the type III capsule locus. Taken together, these findings clarify the phylogenetic position of ST1010 and suggest sequential acquisition of virulence determinants and HLGR prior to its international emergence. IMPACT STATEMENTST1010 GBS has emerged internationally, with colonizing and invasive isolates described in the United States, Dominican Republic, Netherlands, and Italy. Using a core genome phylogeny and targeted detection of genomic regions, we demonstrate that ST1010 shares specific virulence determinants with the CC17 hypervirulent GBS lineage and that HLGR is confined to a specific numerically dominant subclade of ST1010. Our work spotlights the importance of future epidemiologic and genomic surveillance of ST1010 and related lineages. DATA SUMMARYPublicly available genomic data were used from three previously published studies (Laycock KM et al., McGee L et al., Khan UB et al.), as well as a set of newly sequenced GBS genomes from clinical strains originating in New York City (NYC). The corresponding accession numbers and detailed information for all strains are provided in the Table.

Mosquitoes in the Toxorhynchites genus (Theobald, 1901) are specialized predators of container-breeding mosquito larvae, including major Aedes disease vectors. Despite their biological control potential, species boundaries and phylogenetic relationships remain poorly understood. Utilizing country-wide sampling across Panama, we evaluated the taxonomic status and diversity of local Toxorhynchites species. We collected 147 specimens from artificial and natural containers, yielding 97 cytochrome c oxidase subunit I barcodes from 101 sequenced individuals. A Panama-only Neighbor-Joining analysis revealed four distinct, well-supported clusters (I - IV) exhibiting high mean inter-cluster genetic divergence (4.8 - 20.2%), alongside morphological and ecological segregation. A subsequent global Neighbor-Joining analysis incorporating BOLD and GenBank sequences recovered Toxorhynchites as a strongly supported monophyletic group (99.0% bootstrap support) and revealed deep phylogenetic divergence separating Old World Toxorhynchites (Toxorhynchites) (Clades A and B) from New World Toxorhynchites (Lynchiella) (Clades C - G) lineages. Panamanian Clusters I and II formed two distinct groupings within Clade G, nesting as sisters to Subclade G1 characterized by severe taxonomic discordance involving sequences labeled as Tx. moctezuma, Tx. theobaldi, and Tx. rutilus. Cluster III nested within Tx. hypoptes (1.5% divergence) in Clade E, whereas Cluster IV associated with Tx. haemorrhoidalis s.l. (>5.0% divergence) in Clade C. The high degree of taxonomic uncertainty or cryptic diversity uncovered within Clades C, D, and G underscores an urgent need for formal taxonomic revision to accurately delimit species boundaries. Due to its widespread peri-urban distribution, Cluster I (within the Tx. moctezuma s.l. complex) shows the greatest promise for mass-rearing and Aedes biocontrol applications in Panama.

Background: The Human Immunodeficiency Virus and Acquired Immunodeficiency Syndrome (HIV/AIDS) remain a major public health concern in Ghana. Despite sustained progress in treatment and prevention, regional prevalence variations persist, driven by healthcare access, urbanization, and socio-economic factors. This study identifies trends and hotspots to guide effective HIV surveillance and control strategies in Ghana. Methods: A retrospective ecological study was conducted using secondary HIV data confirmed by laboratory testing, from the Ghana District Health Information Management System (DHIMS2) for the period 2020 to 2024. HIV prevalence was calculated as the number of confirmed cases per 100,000 population, using denominators from the Ghana Statistical Service 2021 Population and Housing Census. Spatiotemporal variation in prevalence was visualized using choropleth maps. Global Morans Index examined whether overall spatial dependency existed, followed by local indicators of spatial association (LISA), comprising local Morans I and the Getis-Ord Gi* statistic, to identify local clusters, outliers, and hotspots or coldspots. Results: National HIV prevalence per 100,000 population rose from 0.68 in 2020 to 0.84 in 2024. The highest burden was in the southern and middle belt regions: Western North (2.16), Bono East (1.71), Eastern (1.30), Volta (1.06), and Ahafo (1.01). Northern regions remained consistently low throughout the study period, with Northern (0.32), Upper East (0.26), and Savannah (0.30) recording averages below 0.50 per 100,000. Global Morans Index indicated a dispersed pattern in 2020 (I = -0.43), spatially random pattern between 2021 and 2023, and weak positive spatial association in 2024 (I = 0.22). Conclusions: Regional disparity in HIV prevalence in Ghana is widening, with greater burden concentrated in the more urbanized southern regions. Interventions guided by surveillance data and tailored to specific regions, including strengthened testing infrastructure and a more equitable distribution of health resources, are needed to curb transmission and support HIV in Ghana and the AIDS control programme. Keywords: HIV, AIDS, spatiotemporal analysis, Morans I, Getis-Ord Gi*, Ghana

Monogenean capsalids of the genus Neobenedenia are widespread parasites of wild and farmed marine fish, and represent a great threat to the mariculture of grouper in China. Fishery drug development to screen and find effective compounds to control and prevent the disease is urgent needed, considering the vast production of grouper in China (294 ktons in 2025). Annexins have been discovered in Neobenedenia and other parasites, and marked differences between the parasite annexins and those of the hosts make them potentially attractive drug targets for anti-parasite therapeutics. Herein, we utilized computer-based drug discovery screens using unique Neobenedenia melleni annexin B1 and a database of 1,456,161 small molecules. The 3D structure of annexin B1 was firstly modeled by three different protein prediction tools, namely AlphaFold 3, SWISS-MODEL, and I-TASSER, of which the most accurate protein structure was used as the drug target for the following structure-based virtual screening. In vivo experimental validation of 11 compounds after molecular docking shows that abamectin (Aba) has the most effective anti-Neobenedenia bioactivity at the concentration of 0.16 mg/L as the initial screening concentration. Given its low toxicity to host grouper (24 LC50=0.254 mg/L), abamectin was chose for further investigation. A 24 h bath exposure successfully lowered the parasitic load in infected grouper, yielding an 24 h EC50 of 0.033 mg. To elucidate the anti-parasite mechanism, long-timescale molecular dynamics simulations (1000 ns) of annexin B1 and Aba was conducted, which allowed for atomic and molecular-level analysis of the essential protein motions involved in the interaction of annexin B1 and its substrate. The interaction profile between annexin B1 and abamectin was dominated by hydrophobic contacts and water bridges, involving residues TYR-210, GLU-214, GLU-244, and SER-247, which path a way for further drug optimization.

IntroductionChanges in vector control strategies alter the selection pressures exerted on natural populations of Anopheles gambiae s.l. and may influence the dynamics of insecticide resistance mechanisms. However, the consequences of discontinuing indoor residual spraying (IRS) campaigns on the evolution of Ace-1-mediated resistance remain poorly documented under natural conditions. This study aimed to assess the spatiotemporal evolution of resistance to pirimiphos-methyl following the cessation of IRS and to investigate evidence consistent with the existence of a biological cost associated with Ace-1-mediated resistance. MethodsNatural populations of An. gambiae s.l. were collected between 2017 and 2023 in three districts in Burkina Faso that had undergone IRS campaigns (Kampti, Solenzo, and Kongoussi). Susceptibility tests with pirimiphos-methyl (0.25%) were conducted out following WHO protocols, and a subsample of exposed mosquitoes was genotyped to detect the Ace-1 G119S mutation. Spatiotemporal trends in mortality, allele frequencies and genotypes were analyzed according to the pre-IRS, IRS, and post-IRS periods. The association between the Ace-1 genotype and survival following exposure to pirimiphos-methyl was assessed using logistic regression, while the concordance between phenotypic and molecular indicators of resistance was examined using Spearmans correlation. ResultsThe susceptibility of An. gambiae s.l. populations to pirimiphos-methyl was gradually restored after the discontinuation of IRS at all sites. At the same time, the frequencies of the Ace-1 119S resistance allele declined sharply, particularly in Kampti and Solenzo, while they remained low in Kongoussi throughout the study period. Mosquitoes carrying resistant genotypes had a significantly higher probability of survival after exposure to pirimiphos-methyl than susceptible homozygotes, with resistant homozygotes (RR) exhibiting the greatest survival advantage (OR = 27.62; 95% CI: 6.91-110.45; p < 0.001). A significant negative correlation was observed between the frequency of the Ace-1 119S allele and phenotypic mortality ({rho} = -0.48; p = 0.033), indicating a concordance between the two indicators of resistance. The progressive decline in allele frequencies, the decreasing prevalence of resistant genotypes, and the concomitant restoration of susceptibility are field observations consistent with the existence of biological costs associated with Ace-1-mediated resistance. ConclusionThis study provides field evidence consistent with the existence of biological costs associated with Ace-1-mediated resistance in natural populations of An. gambiae s.l. These results underscore the value of an adaptive resistance management strategy based on alternating selection pressures and could guide future vector control strategies, particularly if indoor residual spraying campaigns or other interventions relying on organophosphates were reintroduced.

Reduced cephalosporin resistance in Enterococcus faecium has traditionally been reported in laboratory mutants and, more recently, in a single clinical ampicillin-susceptible (AmpS) isolate. Herein, we investigated whether this phenotype is widespread by analysing 95 clinical enterococcal isolates (78 AmpS and 17 ampicillin resistant [AmpR]) collected from three hospitals in Spain and Portugal (2009-2025). Low ceftriaxone MICs ([&le;]4 mg/L) were detected in 19/51 (37.3%) AmpS E. faecium and 7/27 (25.9%) E. lactis but in none of the AmpR isolates. Low ceftriaxone MICs were associated with older patient age in both species and with prior ampicillin therapy in E. faecium, but not with other clinical or epidemiological variables. Ceftaroline MICs were consistently low among AmpS isolates, while ceftriaxone and cefotaxime showed greater variability. Low-MIC isolates were distributed across multiple clonal lineages and hospitals and did not share a distinctive resistance or virulence gene profile. PBP5 phylogeny and variation at the psr-pbp5 region separated AmpS from AmpR E. faecium but did not explain variability in ceftriaxone MICs. Five AmpS isolates with reduced ceftriaxone MICs carried chromosomal deletions that included the psr-pbp5 region and genes with diverse cellular functions. Variation in other candidate resistance genes (pbpA, ponA, pbpF, croRS, stpA/stk and murAA) did not consistently explain the MIC differences. These results reveal unexpected heterogeneity in intrinsic cephalosporin resistance in clinical E. faecium and E. lactis and suggest that additional genetic or regulatory mechanisms underlie reduced susceptibility.

Background: Though progress has been made towards universal access to diagnosis and treatment, inequalities in HIV testing are a major obstacle to HIV care in the world, as not everyone has enough access to HIV testing services. Progress is being made towards universal HIV testing and treatment, but inequalities in HIV testing are also an important hurdle to HIV care. Design: Cross-sectional secondary analysis Setting: Nationally representative survey across all 16 administrative regions of Ghana. Participants: 22,058 respondents (15,014 women aged 15-49 and 7,044 men aged 15-59) from the 2022 Ghana Demographic and Health Survey. Primary outcome measure: HIV testing uptake, defined as ever having been tested for HIV and received results (binary: yes/no). Aim: The aim of this study was to examine socioeconomic inequalities and spatial clustering in uptake of HIV testing in Ghana in the 2022 Ghana Demographic and Health Survey (GDHS). Specifically, it considered the level of inequality in wealth, the factors related to these inequalities, and the spatial distribution of HIV testing and factors associated with uptake of HIV testing. Methods: The secondary analysis of cross sectional data was done on 22,058 respondents from all 16 administrative regions of Ghana by using descriptive statistics, Erreygers concentration index, Wagstaff decomposition, spatial autocorrelation techniques, and multilevel logistic regression. Results: The findings showed that there was significant pro-rich inequality in HIV testing, with the majority of inequalities observable not being accounted for by the difference in wealth, but in education level. There was a significant geographic clustering with hotspots in the south, and coldspots in the north. HIV testing uptake was significantly predicted by wealth, education, and the age, sex, marital status and being covered by health insurance Conclusion: The study revealed that socioeconomic and geographical inequalities still exist and have significant impacts on uptake of HIV testing in Ghana. It suggests specific programs such as those aimed at disadvantaged communities, geographic areas, greater access to health insurance, and improved community-based HIV testing services to help ensure equitable access.

The Culex pipiens complex comprises globally distributed mosquito species that serve as vectors for various arboviruses and filariasis. Female mosquitoes within the Cx. pipiens complex are morphologically very similar, making species identification based on morphologically characteristics challenging. To date, Cx. pallens and Cx. pipiens f. molestus have been reported to inhabit the Republic of Korea (ROK). Conversely, Cx. quinquefasciatus, predominantly distributed in tropical regions, has not been documented in the country since a single record in the 1950s. Recently, however, Cx. quinquefasciatus was reported from the Jeju region of the ROK based on molecular identification. This study aimed to re-evaluate the occurrence of Cx. quinquefasciatus in the ROK by conducting molecular identification using the ace-2 and CQ11 markers on Cx. pipiens complex specimens collected nationwide, including from Jeju, where Cx. quinquefasciatus had previously been reported. Our analysis identified only Cx. quinquefasciatus-Cx. pallens hybrid individuals among the Cx. pipiens complex specimens from the Jeju region, highlighting the need to re-evaluate the analytical methods used in previous studies that reported the presence of pure Cx. quinquefasciatus. These findings provide essential evidence for future surveillance of the distribution, introduction, and establishment of Cx. quinquefasciatus and underscore the need for continuous monitoring.

Bacteriophages of the order Crassvirales are currently believed to be the most prevalent dsDNA phages in the human gut virome, yet their global biogeography and genomic diversity remain poorly characterized due to an overrepresentation of industrialized Western studies in public repositories. In this study, we integrated computational metagenomics and molecular approaches to identify and validate the first complete Crassvirales genome from an Egyptian population. De novo assembly and viral profiling yielded a 101,034 bp circular genome (contig k141_108779) predicted to infect the non-industrialized gut symbiont Segatella copri. The genome displays the notable feature of amber stop codon reassignments (NCBI Genetic Code 15), where canonical (TAG) stop codons encode glutamine (Q). This alternative code increases coding density to 91%. Population-level PCR surveillance and Sanger dideoxynucleotide sequencing across 252 individual Egyptian fecal samples, pooled in 10 composites, confirmed the active circulation and local sequence heterogeneity of this lineage within the community. Phylogenomic and intergenomic similarity analysis demonstrated that the isolate shares less than 50% total average nucleotide identity with all recognized type strains. These data establish that this phage constitutes a novel species within a newly proposed genus inside the family Darmviridae. Our findings expand the known geographic distribution of crAss-like phages, highlight translational versatility among Segatella-infecting viruses, and emphasize the importance of expanding virome cohorts to underrepresented regions.

Kinema is a traditional fermented soybean food indigenous to the eastern Himalayan regions of Nepal and India. The fermentation process is primarily mediated by the bacterium Bacillus subtilis, which produces several bioactive compounds and enzymes with potential therapeutic applications. Considering the growing burden of cardiovascular diseases and the need for effective fibrinolytic agents for thrombolytic therapy, this study aimed to extract, partially purify, and evaluate the thrombolytic potential of kinemakinase derived from kinema prepared from white soybeans. Partial purification of the enzyme was achieved using ammonium sulfate precipitation. Thrombolytic activity was assessed in vitro using human blood clots, where three enzyme dilutions demonstrated clot lysis ranging from 66% to 68%, indicating considerable fibrinolytic potential. In silico analyses were also performed to investigate the structural and functional characteristics of the enzyme. The tertiary structure obtained from UniProt was modeled using the Robetta server and refined with GalaxyRefine. Docking with fibrin using ClusPro 2.0 and molecular dynamics simulations using iMODS confirmed favorable interaction and structural stability, while disulfide engineering enhanced protein stability. The findings suggest that kinema-derived kinemakinase may serve as a promising alternative thrombolytic agent, warranting further biochemical characterization and dosage optimization.

Bartonella henselae is a zoonotic pathogen associated with cat-scratch disease. Although multilocus sequence typing (MLST) has been used for strain classification, its resolution for distinguishing between B. henselae isolates remains limited. We herein developed a B. henselae-specific core genome MLST (cgMLST) scheme based on whole-genome sequencing data and examined the genetic and phenotypic diversities of 80 strains derived from cats, humans, mongooses, and masked palm civets. Using the conventional MLST scheme, the 80 strains were classified into nine sequence types (STs), while cgMLST subdivided them into 72 cgSTs, demonstrating a marked improvement in discriminatory power. The cgMLST scheme comprised 1,183 core genes and showed high applicability across the 80 strains. A phylogenetic analysis revealed that ST1, which has been associated with cat-scratch disease, was further subdivided into three major clusters and two singletons, indicating high genetic heterogeneity within this ST. We also found that the bafA subtypes clustered in a manner that was largely consistent with the cgMLST-based phylogenetic structure, suggesting a close relationship between bafA variations and the genomic background of B. henselae strains. In a human umbilical vein endothelial cell proliferation assay, strains belonging to distinct cgSTs exhibited strain-dependent differences in proliferative capacity, which were associated with the bafA subtype classification. Some strains induced focal cell fragmentation and a reduced cell density at a high multiplicity of infection, indicating strain-dependent differences in endothelial cell injury. Collectively, the present results establish a high-resolution cgMLST framework for B. henselae and demonstrate that genetically distinct strains have diverse endothelial cell phenotypes.

Monogenean ectoparasites, particularly Neobenedenia species, cause severe economic losses in mariculture. Here, ectoparasites isolated from cultured hybrid groupers (Epinephelus fuscoguttatus[female] x E. lanceolatus[male]) were confirmed as Neobenedenia melleni based on ITS1 phylogeny. In vivo screening of six structurally diverse compounds identified compound D (CAS No. 206111-37-7), a 5,6-dihydropyridine derivative, as the most effective antiparasitic agent, achieving 76.54% efficacy at 0.5 mg/L in a 90 min bath treatment. Dose-response assays demonstrated that 0.7 mg/L compound D achieved 95.23% antiparasitic efficacy without causing evident tissue damage or cytotoxicity to GF-1 cells. Ultrastructural observation by scanning electron microscopy revealed marked tegumental alterations, including deep fissures and extensive surface folding, in treated parasites. Molecular docking against ten candidate proteins identified {beta}-tubulin as the most favorable docking target, with a binding energy of -6.53 kcal/mol and three hydrogen-bond interactions, suggesting that {beta}-tubulin may be involved in the antiparasitic activity of compound D. Overall, these findings highlight compound D as a promising lead candidate for short-bath therapy against N. melleni and suggest that cytoskeletal disruption through {beta}-tubulin interaction represents a plausible mechanism of action.