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SMART-RNA-Metavirome: a practical RNA metavirome platform compatible with high-throughput sequencing of both short and long reads.

BACKGROUND: The RNA virosphere's extensive diversity and its role in emerging infectious diseases underscore the importance of non-targeted sequencing for identifying unknown or rare pathogens, including co-infections. However, enriching low-abundance viral sequences in RNA metaviromics, particularly in the preparation of cDNA libraries and their compatibility with next-generation sequencing (NGS) and third-generation sequencing (TGS), remains challenging. Therefore, our objective is to develop and systematically assess a practical RNA metavirome methodology specifically tailored for the enrichment of low-abundance viral sequences within samples. METHODS: We developed the SMART-RNA-Metavirome platform, integrating SMART-9n library preparation with NGS and TGS technologies. Total RNA was extracted from two field-collected wild Aedes albopictus pools, along with one laboratory-infected Ae. albopictus pool harboring dengue virus (DENV). This RNA was subjected to reverse transcription using both this optimized protocol and random primer-based methods, followed by high-throughput sequencing on Illumina, Oxford Nanopore, and QitanTech Nanopore technologies. Welch's t-test was employed for comparative analysis of the subsequent RNA metavirome data, specifically to evaluate differences in viral species composition and abundance of viral reads between experimental groups. Furthermore, the effectiveness of this platform was systematically validated via RT-qPCR and SMART-RNA-Metavirome-based Oxford Nanopore sequencing across multiple sample types, including mosquito specimens from DENV-infected Ae. albopictus, serum samples from dengue patients and viral isolates of Japanese encephalitis virus (JEV) and Zika virus (ZIKV). RESULTS: The SMART-RNA-Metavirome platform has been systematically validated to excel in enriching the composition and diversity of the RNA virome (P = 0.04), providing sufficient coverage for the complete reconstruction of viral genomes. When employed in the detection of DENV-infected Ae. albopictus, clinical serum samples, and viral isolates of JEV and ZIKV, this technique exhibits a robust correlation with RT-qPCR (r2 > 0.95). Notably, it demonstrates exceptional sensitivity, ensuring sufficient coverage even in samples of DENV-infected Ae. albopictus with a Ct-value of 35.3, attaining an impressive 99.88% genome coverage. Furthermore, this platform possesses the capability to identify virus species and determine their serotypes. CONCLUSIONS: In our study, the SMART-RNA-Metavirome platform outperforms traditional methods, enriching RNA virome composition and diversity, enabling practical compatibility with both NGS and TGS technologies. It demonstrates significant proficiency in detecting both known and unknown arboviruses, even in low-titer samples such as those from wild mosquitoes and clinical sera. This platform facilitates comprehensive monitoring, risk assessment, and early warning of RNA virus transmissions, enhancing our understanding of RNA virome diversity and ecological patterns.

High-Throughput Nucleotide Sequencing

Recently Evolved, Stage-Specific Genes Are Enriched at Life-Stage Transitions in Flies.

Understanding how genomic information is selectively utilized across different life stages is essential for deciphering the developmental and evolutionary strategies of metazoans. In holometabolous insects, the dynamic expression of genes enables distinct functional adaptations at embryonic, larval, pupal, and adult stages, likely contributing to their evolutionary success. While Drosophila melanogaster (D. melanogaster) has been extensively studied, less is known about the evolutionary dynamics that could govern stage-specific gene expression. To address this question, we compared the distribution of stage-specific genes, that is, genes expressed in temporally restricted developmental stages, across the development of D. melanogaster and Aedes aegypti (A. aegypti). Using tau-scoring, a computational method to determine gene expression specificity, we found that, on average, a large proportion of genes (20%-30% of all protein-coding genes) in both species exhibit restricted expression to specific developmental stages. Phylostratigraphy analysis, a method to date the age of genes, further revealed that stage-specific genes fall into two major categories: highly conserved and recently evolved. Notably, many of the recently evolved and stage-specific genes identified in A. aegypti and D. melanogaster are restricted to Diptera order (20%-35% of all stage-specific genes), highlighting ongoing evolutionary processes that continue to shape life-stage transitions. Overall, our findings underscore the complex interplay between gene evolutionary age, expression specificity, and morphological transformations in development. These results suggest that the attraction of genes to critical life-stage transitions is an ongoing process that may not be constant across evolutionary time or uniform between different lineages, offering new insights into the adaptability and diversification of dipteran genomes.

Animals

Genomic epidemiology of dengue virus 2 and 3 reveals repeated introductions and exportations of several lineages in Colombia.

Dengue fever, a major mosquito-borne viral disease, is transmitted by Aedes mosquitoes and poses a significant global health burden. Despite extensive research, the spatiotemporal dynamics of dengue virus (DENV) lineages in Colombia remain understudied. Here, we analyze 11,443 complete genome sequences from Colombia and the Americas to map the genomic epidemiology of DENV-2 and DENV-3. Phylogeographic reconstruction revealed multiple independent introductions and exportations of the DENV-2 II and III lineages, as well as the DENV-3 lineage III_C.2, underscoring Colombia's critical role as both a source and a sink of viral traffic within the Americas. Antigenic profiling demonstrated distinct clustering of emergent lineages in antigenic space, consistent with immune-escape-driven turnover. These results highlight the necessity of sustained, high-resolution genomic surveillance to guide targeted public health interventions and mitigate dengue transmission across the region.

Dengue Virus

Evolutionary history of Jamestown Canyon virus reveals complex multi-vector ecology.

Jamestown Canyon virus (JCV) is a historically understudied mosquito-borne virus of increasing concern in North America. We generated 658 whole-genome JCV sequences from northeast United States, including 84% (500/597) of all JCV-positive mosquitoes detected in Connecticut from 1997 to 2022. Then, we applied phylodynamic methods to demonstrate how mosquito phenology structures the maintenance and evolution of JCV. Our phylogenetic analyses estimate that JCV was introduced in the Northeast by at least the early 1700s, and the primary introductions of lineages A and B into Connecticut occurred during the mid-1800s to mid-1900s. Further, we estimate that JCV evolves at a rate of ∼3 × 10-5 substitutions per site per year (s/s/y), making it one of the slowest-evolving known RNA viruses, because the virus spends ∼10 months per year in evolutionary stasis while overwintering in mosquito eggs. To investigate ecological drivers of JCV spread in Connecticut, we paired discrete trait and continuous phylogeographic reconstructions with mosquito surveillance data. We estimate that JCV has a low diffusion rate of ∼30-60 km2/year, which is more similar to slow-moving tick-borne viruses than to other mosquito-borne viruses. We found that univoltine Aedes mosquitoes were likely to maintain the virus across years through overwintering in eggs, accounting for its slow evolution and dispersal, while multivoltine mosquitoes contributed to periodic bursts of spatial diffusion and amplification within seasons. We demonstrate the utility of dense sequencing and phylodynamics to disentangle complex transmission cycles, offering a framework for rapidly advancing our evolutionary and ecological knowledge of understudied viruses.

Animals

Mosquitoes provide a transmission route between possums and humans for Buruli ulcer in southeastern Australia.

Buruli ulcer, a chronic subcutaneous infection caused by Mycobacterium ulcerans, is increasing in prevalence in southeastern Australia. Possums are a local wildlife reservoir for M. ulcerans and, although mosquitoes have been implicated in transmission, it remains unclear how humans acquire infection. We conducted extensive field survey analyses of M. ulcerans prevalence among mosquitoes in the Mornington Peninsula region of southeastern Australia. PCR screening of trapped mosquitoes revealed a significant association between M. ulcerans and Aedes notoscriptus. Spatial scanning statistics revealed overlap between clusters of M. ulcerans-positive Ae. notoscriptus, M. ulcerans-positive possum excreta and Buruli ulcer cases, and metabarcoding analyses showed individual mosquitoes had fed on humans and possums. Bacterial genomic analysis confirmed shared single-nucleotide-polymorphism profiles for M. ulcerans detected in mosquitoes, possum excreta and humans. These findings indicate Ae. notoscriptus probably transmit M. ulcerans in southeastern Australia and highlight mosquito control as a Buruli ulcer prevention measure.

Animals

Bioengineered zinc oxide nanoparticles derived from Teucrium polium as a multifunctional platform for anticancer activity, hemocompatibility, larval toxicity and photocatalytic remediation.

In this study, we evaluate the physicochemical properties and the antioxidant, antimicrobial, anticancer, photocatalytic, and larvicidal activities of T. polium-mediated ZnO NPs. The synthesized ZnO NPs were characterized by UV-visible spectroscopy (absorption at 392 nm), FTIR, TEM, and XRD, confirming their successful synthesis. The antioxidant activity of ZnO NPs was evaluated using various assays: DPPH scavenging at 54% (100 µg/mL), ABTS scavenging at 63.3% (100 µg/mL), FRAP scavenging at 61.2% (100 µg/mL), and hydrogen peroxide scavenging at 65% (100 µg/mL), demonstrating concentration-dependent activity. The antibacterial properties were tested against E. coli, P. aeruginosa, K. pneumoniae, and S. aureus, with the largest inhibition zone observed for P. aeruginosa (28.3 mm at 100 µg/mL). Cytotoxicity on MCF-7 cells showed a dose-dependent decrease in cell viability, with values of 75.6%, 44%, 20%, and 8% for concentrations of 25, 50, 75, and 100 µg/mL, respectively. ROS generation and apoptosis were also observed at higher concentrations. The photocatalytic degradation of Methyl Orange was evaluated under UV irradiation, yielding 85% degradation efficiency at pH 3 with a ZnO NP concentration of 50 mg/L. The larvicidal toxicity against Aedes aegypti was significant, with LC50 values of 76.63 µg/mL for III instar and 82.74 µg/mL for IV instar larvae. The results suggest that T. polium-mediated ZnO NPs possess significant potential for therapeutic applications, including antioxidant, antimicrobial, anticancer, photocatalytic, and larvicidal activities, making them a promising candidate for biomedical and environmental applications.

Zinc Oxide

A bunyamwera virus minireplicon system in mosquito cells.

Artificial minigenomes are powerful tools for studying the replication and transcription of negative-strand RNA viruses. Bunyamwera virus (BUN; genus Orthobunyavirus, family Bunyaviridae) is an arbovirus that shows fundamental biological differences when replicating in mammalian versus mosquito cells. To study BUN RNA synthesis in mosquito cells, we developed a bacteriophage T7 RNA polymerase-based minireplicon system similar to that described previously for mammalian cells. An Aedes albopictus C6/36-derived mosquito cell line stably expressing T7 RNA polymerase was established. Viral proteins and artificial minigenomes (containing Renilla luciferase as a reporter) were transcribed and expressed in these cells from transfected T7 promoter-containing plasmids. Transcription of the minigenome required two viral proteins, the nucleocapsid protein N and the RNA-dependent RNA polymerase L, a situation similar to that in mammalian cells. However, unlike the situation in mammalian cells, the viral polymerase was not inhibited by the viral nonstructural protein NSs. We also report that promoter strength is different for vertebrate versus invertebrate cells. The development of this system opens the way for a detailed comparison of bunyavirus replication in cells of disparate phylogeny.

Aedes

Comparative dynamics of Japanese encephalitis virus adaptation in porcine macrophages and insect cells.

BACKGROUND: Japanese encephalitis virus (JEV) is a zoonotic mosquito-borne Orthoflavivirus that circulates primarily in birds and pigs. Previous observations of vector-free transmission between pigs indicates the possibility of single-host cycling in swine. Therefore, the aim of this work was to investigate the evolutionary pressure of single host cycling using a relevant primary cell culture model. METHODS: To investigate whether such single-host cycles affect viral infectivity, fitness and genomic adaptations, two strains and a reverse genetic cDNA-derived clone of JEV were serially passaged 12 times in primary porcine monocyte-derived macrophages (MDMs), in Aedes albopictus-derived C6/36 cells, and alternately between both cell types. Next-generation sequencing analysis was used to identify selected single nucleotide variants (SNVs) and haplotypes. Phenotype-to-genotype connections were confirmed using reverse genetics. RESULTS: For all viruses, serial passaging in MDMs - but not in C6/36 cells - led to a rapid increase in relative infectivity toward MDMs, accompanied by reduced plaque sizes in porcine endothelial cells. In contrast to C6/36 cells, MDM imposed a strong selective pressure, rapidly favoring selection of many SNVs and viral haplotypes. In addition, we identified a dominant selection of mutants with glutamic acid to lysine substitutions at positions 49 or 138 in the E protein, which explained the small plaque phenotype and caused viral sensitivity to heparin-mediated inhibition of attachment, indicating enhanced virus binding to glycosaminoglycans (GAG). The E138K mutant also explained the increased relative infectivity for MDM. CONCLUSION: This work demonstrates a high evolutionary pressure on JEV in MDM causing rapid selections of minor haplotypes. Furthermore, the efficient selection of E49K and E138K SNV, which were responsible for the phenotype, are likely caused by a selective pressure for GAG binding, observed in vitro with other mammalian cells.

Animals

Autochthonous chikungunya virus (CHIKV) outbreak in the province of Modena, Emilia-Romagna region, Italy, August to October 2025: epidemiology, clinical features and virological findings.

Between 10 August and 31 October 2025, 343 cases (306 confirmed and 37 probable) of symptomatic autochthonous chikungunya virus (CHIKV) infection were notified in eight municipalities in the province of Modena, Emilia-Romagna region, northern Italy. The infection was diagnosed by detection of CHIKV RNA in blood and urine samples, and by detection of IgM antibodies against CHIKV. Most common symptoms were arthralgia (n = 326) and fever (n = 317). No fatalities were reported. Chikungunya virus RNA was detected in 39 (14.8%) of 263 pools of Aedes albopictus mosquitoes, thereby confirming local vector-borne transmission. Sequences from 11 case samples and two mosquito pools were whole genome sequenced. The viral strain belonged to the East/Central/South African (ECSA) lineage 2 and was closely related to the strains circulating in Reunion Island in 2024-25. The public health response included rapid vector control measures, reinforced epidemiological surveillance and multidisciplinary coordination between public health authorities and clinicians, laboratories and entomologists. This was one of the largest autochthonous CHIKV outbreaks ever recorded in a temperate European region. It highlights the critical importance of integrated, multidisciplinary preparedness and response for arboviral threats in non-endemic areas.

Humans

RT-RPA-Assisted CRISPR/Cas12a-Based Isothermal Detection of Chikungunya Virus.

Chikungunya virus (CHIKV) is transmitted through the bite of Aedes mosquitoes, specifically A. aegypti and A. albopictus. CHIKV belongs to the alphavirus with a positive-sense ssRNA genome of 11-12 kb size. The virus has been reported from various geographical regions across the globe. Chikungunya fever is an acute febrile illness, which, if left untreated, may develop into chronic arthralgia that may persist for several months or acute encephalitis syndrome. Therefore, early diagnosis of CHIKV is crucial to initiate prompt supportive treatment. Laboratory diagnosis of CHIKV typically relies on serological tests such as IgM antigen capture ELISA and molecular methods including RT-PCR or qRT-PCR. However, both these methods are not viable in peripheral settings. This chapter highlights recent advancements in molecular detection techniques for CHIKV, specifically isothermal detection methods that eliminate the requirement for complex instruments. The detection is facilitated by RT-RPA and CRISPR/Cas12a endonuclease. The assay offers advantages over existing methods such as rapid and early detection, and eliminates cross-over contamination, ultra-sensitivity, high specificity, and ease of result interpretation.

Chikungunya virus

RNA viruses in sylvatic mosquitoes and phlebotomine sand flies from Alto Pantanal, Mato Grosso, Brazil 2019.

The Pantanal biome harbors exceptional biodiversity but has been increasingly impacted by climate change and human activities. This region is considered a high-risk zone for zoonotic spillover, making viral studies in sylvatic mosquitoes and other invertebrates indispensable, as these vectors are involved in the transmission of pathogens of public health concern. This study aimed to describe viral genomes identified in Aedes spp., Ochlerotatus sp., Mansonia sp., Phlebotomus sp., Psorophora spp., and Anopheles spp. dipterans collected in March and June 2019, in Pirizal and Porto São Luiz, Alto Pantanal, Mato Grosso State, Brazil. Diptera specimens were pooled by genera, and nucleic acids were extracted, followed by library preparation and sequencing on the Illumina NextSeq 500/550 platform. A total of 39 putative viral sequences were recovered, including 23 potentially novel viruses. Coding-complete genomes were identified from Virgaviridae (n=1), Rhabdoviridae (n=1), and Metaviridae (n=1), as well as seven coding-complete segments from Partitiviridae (n=4) and Solemoviridae (n=3). Additionally, 29 partial genomes were recovered from Partitiviridae (n=7), Metaviridae (n=6), Chuviridae (n=2), Sedoreoviridae (n=1), Nodaviridae (n=3), Tombusviridae (n=2), Phasmaviridae (n=2), Flaviviridae (n=3), Virgaviridae (n=1), and Solemoviridae (n=2). Viral characterization in Diptera specimens has gained increasing importance with the advancement of metagenomic approaches, which contribute to global One Health initiatives by providing data that may support the prediction and prevention of future viral spillover events.

Animals

Detection and genomic characterization of a travel-associated ECSA lineage chikungunya virus infection in Mexico.

BACKGROUND: In 2013, chikungunya virus (CHIKV), a re-emerging Aedes-borne virus, was introduced into the Americas. This led to synchronous epidemics across the region associated mainly with the Asian lineage, which eventually subsided. Resurgent outbreaks have been recorded since, principally in South America, largely driven by the East-Central-South-African (ECSA) lineage. In 2025, more than 300,000 CHIKV suspected cases were reported in Brazil and Cuba. CASE SUMMARY: In November 2025, a healthy adult male traveling from Cuba arrived in Merida, Mexico, and shortly after presented febrile symptoms consistent with an arboviral infection. CHIKV infection was diagnosed by RT-qPCR. Though the infection was mild, the patient developed a rash on the abdomen and neck that persisted for up to a month, with further inflammation of the joints of the left leg. Phylogenetic analysis of the viral genome indicated placement within the ECSA lineage, clustering with other contemporaneous virus genomes sampled from Brazil that belong to a recently described clade II within the country, in which viral genomes from Cuba also cluster. CONCLUSION: We identify a travel-associated ECSA lineage CHIKV case in Mexico. This viral lineage has not previously been detected in the country. This finding highlights the risk for subsequent local transmission and is consistent with reports of the presence of this lineage in Cuba. Ten years since the last CHIKV epidemic in Mexico, strengthened surveillance is required to anticipate potential local outbreaks within the region.

ECSA

The Bunyamwera orthobunyavirus Gc glycoprotein head and stalk drives an infectious virion assembly pathway specific for the insect host.

The Orthobunyavirus genus of arthropod-borne segmented RNA viruses comprises important pathogens including the human-infecting Oropouche virus and ruminant-infecting Schmallenberg virus (SBV). The prototypical Bunyamwera orthobunyavirus (BUNV) possesses envelope-embedded glycoprotein Gn-Gc tripodal spikes, of which the ectodomains mediate virus entry, while endodomains interact with nucleoprotein (NP) enwrapped genome segments driving virion assembly. Interestingly, BUNV Gc head/stalk domains are redundant for virus growth in mammalian cells, consistent with isolations of SBV from ruminants bearing head/stalk deletions. However, these domains appear strictly maintained in orthobunyavirus isolations from arthropods in nature. To investigate the molecular mechanism that underlines this discrepancy, we compared the multiplication characteristics of wildtype BUNV (BUNV-WT) with a Gc head/stalk deleted BUNV (BUNV-∆7). In mammalian cells BUNV-WT and BUNV-∆7 grew to equivalent titres, whereas in insect cells BUNV-∆7 titres were 1000-fold lower and strikingly produced no virions following blood meal infection of Aedes mosquitoes. To understand this insect-specific restriction in virion production, we showed the intracellular abundance of BUNV-WT and BUNV-∆7 Gc and NP components were equivalent, suggesting the deletion impacted post-translational stages of the infection cycle. To explore this, we investigated Gc and ∆7-Gc interactions during BUNV-WT and BUNV-∆7 infections of both insect and mammalian cells by co-immunoprecipitation and multiplex mass spectrometry, revealing ∆7-Gc exhibited markedly reduced NP interactions in insect cells, potentially indicating reduced segment interactions during assembly. We hypothesize that the Gc head/stalk performs an insect cell-specific role in segment recruitment during virion formation, and that maintenance in nature of full-length Gc is due to this essential role in the insect host.

Animals

Global molecular and serological evidence of dengue and chikungunya infection: a systematic review and meta-analysis of 158,608 tested participants.

INTRODUCTION: Dengue virus (DENV) and chikungunya virus (CHIKV) are Aedes-borne arboviruses with overlapping clinical manifestations, shared vectors, and substantial diagnostic challenges in co-endemic settings. This systematic review and meta-analysis synthesized published evidence on molecular detection, serological positivity, and DENV-CHIKV dual positivity/co-infection in human clinical, surveillance, and community-based study populations. CONTENT: Following PRISMA 2020 guidance, five bibliographic databases (PubMed/MEDLINE, Scopus, Web of Science, ScienceDirect, and Google Scholar) and supplementary grey-literature/preprint sources were searched for English-language studies published from 1 January 1980 to 31 December 2024. No prospective PROSPERO or OSF protocol registration was available. Eligible records reported extractable numerators and denominators for DENV and/or CHIKV in humans using recognized molecular or serological assays. A total of 196 studies comprising 158,608 tested or suspected participants were included in the extraction table. The pooled CHIKV estimate was 14.0 % (95 % CI: 12.0-16.4; I2=97.5 %), with molecular and serological estimates of 9.8 and 15.7 %, respectively. The pooled DENV estimate was 13.8 % (95 % CI: 10.9-17.3; I2=99.0 %), with molecular and serological estimates of 13.1 % (95 % CI: 7.9-21.0) and 14.3 % (95 % CI: 10.2-19.8), respectively. DENV-CHIKV dual positivity/co-infection was 52.9 % (95 % CI: 48.7-57.1) among studies that tested and reported both outcomes. Country-level estimates varied widely and should be interpreted as summaries of available studies rather than nationally representative burden estimates. Funnel-plot asymmetry was statistically significant in DENV analyses but not in the overall CHIKV analysis. SUMMARY: Available evidence indicates extensive but highly heterogeneous DENV and CHIKV positivity across selected clinical and surveillance populations. The pooled estimates should be interpreted cautiously because of substantial between-study heterogeneity, diagnostic variability, outbreak-period sampling, and uneven geographic representation. OUTLOOK: The findings support integrated arboviral surveillance, multiplex diagnostics, and vector-control preparedness in co-endemic regions.

Humans

A new potential mosquito-borne virus: detection of Human-derived Jingmenvirus in several-species of mosquitoes from Yaoundé, Cameroon.

BACKGROUND: Tick-borne Jingmenviruses are becoming an increasing arbovirus concern due to the rising number of reported infections in humans and animals, as well as their wide geographic distribution. The involvement of other hematophagous arthropods as vectors of Jingmenviruses is still unknown. METHODS: Mosquitoes were sampled in two different biotopes in Cameroon (Yaoundé and Garoua) during the rainy and the dry seasons in 2022 and 2023. Metatranscriptomics Next Generation Sequencing was conducted using Illumina technology. Viral sequences detection revealed the presence of several contigs with high sequence identity to a human-derived Jingmenvirus (HdJV) previously discovered in plasma from an individual from Yaoundé, Cameroon. A draft viral genome was constituted for each Jingmenvirus-positive samples. Maximum likelihood phylogenetic reconstructions were used to position mosquito-associated viruses within the diversity of Jingmenviruses. Statistical analyses were conducted to estimate the prevalence of infected mosquitoes and the effect of different variables (region, season, year, mosquito species) on Jingmenvirus detection. RESULTS: HdJV was identified during the dry and the rainy seasons in 4 species of mosquitoes: Aedes albopictus, Culex quinquefasciatus and Culex wansoni from Yaoundé, and Anopheles gambiae s.l. from Garoua. The overall prevalence of HdJV-infected mosquitoes was estimated to 0.90% [0.41-1.69]; and the unique variable significantly associated with HdJV detection was the sampling area: Yaoundé showed the highest prevalence (2.29% [0.95-4.68]) compared to Garoua (0.18% [0.01-0.79]). Mosquito-associated Jingmenviruses shared a high nucleotide identity (between 98.64-100% according to the segment) and clustered in the same clade in the phylogenetic analysis, that they belong to the same viral species circulating in different mosquito species. The viral genome shared between 96.4% and 98.9% nucleotide identity with a HdJV detected in the plasma of a patient suffering from febrile illness originating from the same area, suggesting the possible involvement of mosquitoes as vectors of arboviral Jingmenviruses in human infections. CONCLUSIONS: This finding provides new insights into the ecology and transmission dynamics of Jingmenviruses, highlighting mosquitoes as potential vectors, alongside ticks, in the zoonotic transmission of this virus group.

Jingmenvirus

Frequent introductions and climate suitability drive increasing dengue risk in Florida.

In recent years, detection of local dengue cases in Florida have increased in both frequency and geographical extent. From 2022 to 2024, consecutive outbreaks in Miami-Dade County were mainly caused by a single lineage of dengue virus (DENV) serotype 3, prompting questions about changing epidemiology and a transition towards endemicity. In this study, we used mathematical modeling and genomic epidemiology to reveal the spatiotemporal dynamics and drivers of local dengue cases in Florida. We found that annual clusters and outbreaks were caused by frequent short-lived DENV introductions, primarily from the Caribbean, and did not find evidence for local trans-seasonal DENV lineage persistence. Further, we show that the climate-driven increases in local suitability for Aedes aegypti transmission and travel-associated cases were the greatest risk factors for outbreaks in Miami-Dade and the geographic expansion of dengue in Florida. Overall, while we do not yet find evidence for endemicity, we demonstrate how climatic trends are enhancing the local public health risk caused by dengue in Florida.

Journal Article

Bunyamwera bunyavirus nonstructural protein NSs is a nonessential gene product that contributes to viral pathogenesis.

Bunyamwera virus (family Bunyaviridae, genus Bunyavirus) contains a tripartite negative-sense RNA genome. The smallest RNA segment, S, encodes the nucleocapsid protein N and a nonstructural protein, NSs, in overlapping reading frames. We have generated a mutant virus lacking NSs, called BUNdelNSs, by reverse genetics. Compared with the wild-type (wt) virus, BUNdelNSs exhibited a smaller plaque size and generated titers of virus approximately 1 log lower. In mammalian cells, the mutant expressed greatly increased levels of N protein; significantly, the marked inhibition of host cell protein synthesis shown by wt virus was considerably impaired by BUNdelNSs. When inoculated by the intracerebral route BUNdelNSs killed BALB/c mice with a slower time course than wt and exhibited a reduced cell-to-cell spread, and titers of virus in the brain were lower. In addition, the abrogation of NSs expression changed Bunyamwera virus from a noninducer to an inducer of an interferon-beta promoter. These results suggest that, although not essential for growth in tissue culture or in mice, the bunyavirus NSs protein has several functions in the virus life cycle and contributes to viral pathogenesis.

Aedes