Wild Turkeys (Meleagris gallopavo, hereafter turkeys), an important North American game species, have experienced declines throughout their eastern range. Growing concern over turkey population sustainability has renewed interest in investigating potential disease threats. We conducted pathogen surveillance in turkeys in three North Carolina, USA ecoregions-Mountains, Piedmont, and Coastal Plain-in 2020-22 to provide baseline data relevant to the southeastern USA. We collected samples from 586 live free-ranging turkeys plus 22 recaptured individuals (n=608: 194 males, 414 females; 159 juveniles, 449 adults) to test for exposure to or infection with selected pathogens. Molecular testing revealed infections with Haemoproteus spp. (57%), lymphoproliferative disease virus (LPDV; 46.8%), Mycoplasma spp. (39.8%), Leucocytozoon spp. (8.8%), and reticuloendotheliosis virus (REV; 3.4%). We detected antibodies to Toxoplasma gondii (21.3%), West Nile virus (WNV; 15.4%), and avian influenza virus (2.0%). No turkey coronavirus, Plasmodium, Borrelia, or Salmonella infections were detected. There were no prevalence differences between sexes, except for REV (females=5%, males=1%). Prevalence was higher in adults than in juveniles for LPDV (adult=52%, juvenile=33%), WNV (adult=19%, juvenile=6%), Haemoproteus (adult=60%, juvenile=49%), T. gondii (adult=24%, juvenile=14%), and Leucocytozoon (adult=11%, juvenile=3%). Prevalence of LPDV differed significantly across ecoregions, with the highest prevalence in the Piedmont (62%), followed by the Mountains (51%) and the Coastal Plain (27%). Prevalence of WNV antibodies was higher in the Piedmont (24%) than in the Mountains (8%). Haemoproteus and Leucocytozoon prevalence increased over a regional gradient, with detections of 24% and 0% in the Mountains, 65% and 6% in the Piedmont, and 85 and 21% in Coastal Plain, respectively. Mycoplasma spp. prevalence was higher in the Mountains (45%) and the Coastal Plain (47%) than in the Piedmont (27%). Our data highlighted sex-, age-, and region-based differences in prevalence for several pathogens, thereby enabling managers to tailor management strategies and researchers to investigate effects of these pathogens on turkey survival and movement.

While diabetes-related complications have been widely investigated, the burden of infectious diseases across the diabetes spectrum remains relatively understudied. We developed a Bayesian approach to compare infection risk across 9,476 patients with type 1 diabetes (T1D), 74,270 with type 2 diabetes (T2D), and 32,095 with prediabetes. Patients with T1D, T2D, and prediabetes had multifold increased risk for all organ system- and pathogen-based composite infection outcomes. We also quantified risk for 1,401 individual infection outcomes, finding increased risk for most infections among patients with either T1D, T2D, or prediabetes. Patients had increased risk for well-established diabetes-associated infections (e.g., mucormycosis) and less commonly associated infections (e.g., West Nile Virus encephalitis). Finally, we found disparities in risk across sociodemographic subgroups (i.e., age, sex, ethnicity, ancestry, and insurance status). Our comprehensive findings advance previous research by quantifying risk for wide-ranging infection outcomes across diverse patients with T1D, T2D, and prediabetes through an innovative Bayesian approach.

West Nile virus (WNV) is an endemic arboviral infection in the United States that has undergone phylogenetic evolution since its introduction 25 years ago. An integrated vector-human-pathogen study was conducted in the summer of 2023 to unearth contemporary Culex quinquefasciatus habitat patterns and human transmission spillover foci in South Carolina, a state with historically little WNV data. A serosurvey revealed WNV seroprevalence 10 times the national average (22% versus 2%, respectively), with unusual epidemiologic risk factors. Female Culex quinquefasciatus WNV positivity was low (2.7%), with viral phylogenetics 100% homologous to the WN02 clade. Mosquito vectors clustered in affluent urban neighborhoods with greater tree canopy cover and abundant waterbodies. Culex quinquefasciatus abundance was greatest when climate variance was nominal in the 72 hours preceding collection. An unusual bimodal mosquito temporal pattern was observed, reflecting changing climate patterns. The present comprehensive WNV study reveals emerging transmission factors as WNV continues to evolve and persist in the southeastern United States.

Neuroinvasive West Nile Virus Infection in Kidney Transplantation: Three Case Reports.

West Nile virus (WNV) is primarily transmitted by the bite of Culex mosquitoes, but other mechanisms, such as blood transfusion, have also been described. Since its identification in the Americas in 1999, WNV has circulated consistently in the United States of America (USA); however, although WNV has been detected in humans in South America, no major outbreaks have occurred in more than 20 years. One of the hypotheses to explain this difference is the underdiagnosis of the infection. In Mexico, nine isolated cases have been officially reported since 2003 despite its proximity to the USA. In this study, we aim to demonstrate the circulation of WNV in blood donors from a northern border city of Mexico. Between August and September of 2023, 86 serum samples from volunteer blood donors were collected to determine anti-WNV Immunoglobulin (Ig) G using a commercial enzyme-linked immunosorbent assay (ELISA) kit. In a subgroup of 44 samples, anti-WNV IgM was determined. To corroborate the IgM results, nucleic acid amplification tests (NAAT) were performed to determine RNA of WNV, Dengue and Zika. The participants were questioned about the history of travel to the USA; all of them were residents of the city of Nogales, Sonora, located on the border with the state of Arizona. For the comparison of seronegative and seropositive donor groups, the Chi-square test and Mann-Whitney U test were used for qualitative and quantitative variables, respectively. Additionally, a spatial analysis of seropositive cases was conducted. One sample was reactive to anti-WNV IgM and IgG; however, all NAAT results were negative. In addition, 19 samples were reactive for IgG, and no statistically significant differences were found between the groups. Seropositive cases showed a geographic pattern of clustering on the outskirts of the city, in areas with low population density. Our results strongly suggest recent WNV circulation among residents from the northern border of Mexico. The lack of differences regarding the history and frequency of travel to the USA suggests that contact with the virus occurs in Mexico and that the low reported circulation in the region represents an underdiagnosis of the disease.

Orthoflaviviruses, such as tick-borne encephalitis virus (TBEV) and West Nile virus (WNV), can cause severe neurological disease and remain without specific antiviral treatments. We found that orthoflavivirus envelope (E) and non-structural protein 1 (NS1) interact with heat shock protein 70 (Hsp70) chaperones, key regulators of protein homeostasis and existing cancer drug targets. We examined how Hsp70 and endoplasmic reticulum–resident BiP contribute to viral protein secretion and infectivity of tick and mosquito-borne orthoflaviviruses. Targeting the Hsp70 nucleotide-binding domain with small-molecule inhibitor YM-1 significantly reduced infectivity of multiple orthoflaviviruses, while substrate-binding domain inhibitor PES-Cl specifically impaired NS1 secretion of tick-borne orthoflaviviruses. Protein degradation inhibitors restored NS1 expression in BiP-deficient cells but failed to rescue NS1 secretion. These data indicate that while BiP is essential for secretion of tick-borne orthoflavivirus NS1, it is not required for infectivity. The antiviral effect of YM-1 likely reflects inhibition of other chaperones or additional cellular targets.

Seroprevalence of four zoonotic viral diseases in camels in Qatar.

Validation of prototype virus inactivation from seven virus families of pandemic potential with a novel low-cost, field-deployable RNA extraction and storage method.

Metagenomic surveillance of emerging viruses in mosquito populations from high-risk regions of Iran.

Japanese encephalitis virus (JEV), first identified in 1935, continues to be a major threat to human health, especially in the Asia-Pacific region, where it remains prevalent. JEV, a neurotropic flavivirus, spreads through Culex tritaeniorhynchus mosquito bites and causes severe brain infections with high morbidity and mortality rates. Despite the availability of vaccines, no licensed anti-JEV drugs exist. This review provides a comprehensive overview of the epidemiology, structural and nonstructural proteins, and pathogenesis of JEV and explores potential drug targets. This study highlights both conventional and nonconventional drug targets, with a focus on nonstructural JEV proteins, which may hold promise for therapeutic development. This review also discusses drug targets shared by JEV and other flaviviruses, such as dengue, Zika, and West Nile virus, which reveal common pathways for viral entry and replication, along with distinct mechanisms specific to JEV. Key receptor interactions, including DC-SIGN, TAM receptor, sialic acid, LDLR, and CLEC5A interactions, are involved in JEV transmission and immune evasion. Additionally, the NMDA receptor has been identified as a critical player in JEV pathogenesis, suggesting new opportunities for neuroprotective therapies. A major obstacle in JEV drug development is the blood-brain barrier (BBB), which hinders the delivery of therapeutic agents to the central nervous system (CNS). Recent research has emphasized the need for innovative drug delivery systems that can cross the BBB, reducing viral replication and neural damage. While clinical trials with traditional antivirals have yielded mixed results, live attenuated and inactivated vaccines have shown promise in preventing JEV infection. Additionally, nucleic acid-based therapies, including microRNAs and short hairpin RNAs (shRNAs), are emerging as potential treatments, with nanoparticle-based delivery systems offering solutions to overcome BBB challenges. This review underscores the need for an integrated approach, including improved vaccines, targeted drug delivery strategies, and novel therapeutics, to effectively combat JEV infections on a global scale.

Human autoantibodies against type I interferons in severe viral disease.

Concurrent West Nile virus neuroinvasive disease and progressive multifocal leukoencephalopathy in advanced HIV

Spreading speed of a nonlocal dispersal West-Nile virus epidemic model with shifting habitats

Mosquito-borne diseases, including malaria, dengue, Zika, and West Nile virus, continue to pose significant global health threats, necessitating the development of effective personal protection measures (1,35). While synthetic repellents such as N,N-diethyl-meta-toluamide (DEET) have served as the gold standard for decades, increasing concerns regarding their neurotoxicity, skin irritation, and environmental persistence have shifted consumer preference toward plant-based alternatives (10,21). This review critically examines the efficacy and development of herbal sprays, focusing on the botanical profiles and repellent properties of guava leaf extract (Psidium guajava), castor oil (Ricinus communis), and marigold extract (Tageteserecta) (3–5). Furthermore, we analyze the role of secondary metabolitesterpenoids, alkaloids, and phenolics—in disrupting insect host-seeking behavior(6,7). The article outlines standardized testing methodologies (WHO/ASTM) and essential safety protocols required to transition these traditional remedies into reliable, commercial-grade biopesticides (1,17). Challenges such as high volatility and the need for advanced formulation techniques like microencapsulation are discussed as the way forward for sustainable insect management (26,31).

Arboviruses transmitted by mosquitoes pose a global health threat, causing diseases ranging from mild fevers to severe encephalitis and hemorrhagic fevers. Despite their growing impact, arbovirus research is hindered by biosafety constraints and the need of specialized BSL-3 insectariums. To circumvent these challenges, mosquito-derived cell lines have become indispensable tools for investigating virus-vector interactions. However, most available cell lines originate from Aedes and Anopheles spp., creating a critical research gap for other key vectors such as Culex spp. Although a few cell lines were previously established, they did not represent primary transmitters of West Nile virus (WNV) and other emerging arboviruses in Europe, such as Culex pipiens. To address this gap, the current study aimed to characterize two recently established Culex pipiens cell lines: CPE/LULS50 (Culex pipiens pipiens & molestus) and CPL/LULS56 (Culex pipiens molestus) in more detail including testing their virus susceptibility, antiviral RNAi response, and possible presence of insect-specific viruses. The replication of arboviruses from three clinically relevant families (Flaviviridae, Peribunyaviridae, and Togaviridae), as well as insect-specific viruses, was observed in both CPE/LULS50 and CPL/LULS56 cell lines. Furthermore, small RNA profiling revealed production of virus-specific small interfering RNA (siRNA) in both cell lines for all tested viruses. Interestingly, virus-specific PIWI-interacting RNA (piRNA) was only detected for the Peribunyaviridae. The current study demonstrates that the CPE/LULS50 and CPL/LULS56 cell lines are suitable candidates to facilitate research into Culex-specific virus-vector interactions, ultimately contributing to mitigation of the impact of Culex-borne arboviruses on public health.

Arboviral infections, notably those caused by West Nile virus (WNV), Dengue virus (DENV), Zika virus (ZIKV), and Chikungunya virus (CHIKV), represent a growing global health concern due to their expanding geographic distribution and evolving clinical spectrum. While systemic and neurologic complications are well recognized, cutaneous manifestations are frequently overlooked despite their diagnostic and prognostic value. These viruses, transmitted primarily by Aedes or Culex mosquitoes, share overlapping symptoms such as fever, headache, arthralgia, myalgia, and malaise but can often be distinguished by specific dermatologic features. WNV may present with a subtle truncal exanthem; DENV is characterized by flushing, petechiae, and the 'white islands in a sea of red' pattern; ZIKV typically is associated with a fine, pruritic maculopapular rash with palmar-plantar involvement, edema and erythema of the malar region of the face, and conjunctival injection; CHIKV frequently shows an asymptomatic or pruritic macular or maculopapular rash that can result in pigmentary changes mostly on the face and mucocutaneous involvement. Other arboviral infections such as Usutu virus (USUV) and Toscana virus (TOSV) more rarely exhibit cutaneous signs. These include transient macular or urticarial rashes, conjunctivitis, and aphthous-like ulcers on the palate during early or prodromal phases. This review synthesizes current clinical, virologic, and pathophysiologic insights into these cutaneous presentations, highlighting their diagnostic relevance, underlying mechanisms, and implications for dermatologic and interdisciplinary care. Recognizing these dermatologic signs can significantly aid early diagnosis and improve patient management in endemic and outbreak settings.

West Nile virus (WNV) is an endemic arthropod-borne virus that has routinely caused seasonal outbreaks in the United States since it was first detected in 1999. While phylogenetic studies have shown how WNV has diversified and undergone genotype replacement since introduction, more geographically focused studies are needed to understand intricate transmission dynamics at local and regional scales. In this study, we validate the IDT xGen WNV panel, a novel single reaction amplicon-based Next-Generation Sequencing approach, to generate high-quality WNV genomes and compare it to the "Primal Scheme" assay for WNV, a common amplicon sequencing strategy. By generating >250 genomes from mosquito pools, we show that the IDT xGen WNV panel generated coding-complete and accurate WNV genomes when compared to the current sequencing approaches. Additionally, we used this approach to generate 100 coding-complete WNV genomes from surveillance pools of mosquitoes collected in Nebraska during the 2023 outbreak. Our discrete phylogeographic analysis revealed substantial genetic diversity in WNV genomes from 2023 with minimal clustering across the state. This study demonstrated the utility of a single reaction amplicon-based sequencing approach to generate quality WNV genomes from routine surveillance samples and characterize WNV transmission dynamics in a high-incidence setting.

The rise of flaviviral diseases, including West Nile virus (WNV), presents a growing threat to global public health and underscores the urgent need for new therapeutic strategies. The non-structural protein 3 helicase (NS3h) of the Orthoflavivirus genus, including WNV, is essential for viral replication and a promising antiviral target. Previously [Roy etal., Nucleic Acids Research, 52 (13), 2024, 7447-7464], we showed that the motif VI loop (VIL) in WNV NS3h functions as a nucleotide valve, regulating ADP affinity during hydrolysis. In this study, we uncover an ATP-dependent coupling between nucleotide affinity at motif VIL and RNA affinity at motifs IVa and V, suggesting a coordinated mechanism of ssRNA translocation. Using microsecond-scale all-atom molecular dynamics simulations of hydrolysis-cycle intermediates, we find that key VIL residues (R461, R464) correlate strongly with RNA phosphate affinity of motif V. Structural analyses reveal an ATP-sensitive interaction between E413 (motif V) and R461 (motif VIL) that modulates the conformation of the motif V 310-helix, thereby influencing RNA binding. This dynamic interaction is lost in catalytically deficient VIL mutants, which have been experimentally shown to impair hydrolysis and attenuate viral replication. These findings provide mechanistic insights into NS3h function and identify new opportunities for structure-based antiviral design.

West Nile virus (WNV) is a zoonotic mosquito-borne Flavivirus, with bird populations reservoirs. Although often asymptomatic, infection in humans can cause febrile symptoms and, more rarely, severe neurological symptoms. Previous studies assessed environmental drivers of WNV infections, but most overlooked areas with potential WNV circulation despite no reported human case, and mixed mechanisms affecting hosts vs. vectors. Our objective was to generate a WNV Bird Risk Index (BRI) mapping the potential of WNV circulation in bird communities across Europe. We first used a bird traits-based model to estimate WNV seroprevalence in European wild bird species and identify eco-ethological characteristics associated with it. This allowed us to build a map of the WNV BRI that showed a strong spatial heterogeneity across Europe. To validate this metric, using a Besag-York-Mollie 2 spatial model in a Bayesian framework, we showed a positive association between the BRI and the number of years with notified WNV human cases between 2016 and 2023, at the NUTS administrative region scale. To conclude, we provide a map quantifying the suitability for WNV to circulate in the bird reservoir. This allows to target surveillance efforts in areas at risk for WNV zoonotic infections in the future.

Surveillance-based insights into mosquito-borne disease trends: Implications for public health in Poland and Europe (2018-2024).