Culex-Transmitted Diseases: Mechanisms, Impact, and Future Control Strategies using Wolbachia.

Mosquitoes of the Culex genus are responsible for a large burden of zoonotic virus transmission globally. Collectively, they play a significant role in the transmission of medically significant diseases such as Japanese encephalitis virus and West Nile virus. Climate change, global trade, habitat transformation and increased urbanisation are leading to the establishment of Culex mosquitoes in new geographical regions. These novel mosquito incursions are intensifying concerns about the emergence of Culex-transmitted diseases and outbreaks in previously unaffected areas. New mosquito control methods are currently being developed and deployed globally. Understanding the complex interaction between pathogens and mosquitoes is essential for developing new control strategies for Culex species mosquitoes. This article reviews the role of Culex mosquitos as vectors of zoonotic disease, discussing the transmission of viruses across different species, and the potential use of Wolbachia technologies to control disease spread. By leveraging the insights gained from recent successful field trials of Wolbachia against Aedes-borne diseases, we comprehensively discuss the feasibility of using this technique to control Culex mosquitoes and the potential for the development of next generational Wolbachia-based control methods.

Dengue outbreaks in Brazil and Latin America: The new and continuing challenges.

Dengue is one of the most common zoonoses in Latin America. In 2023, Brazil registered a total of 1,658,814 suspected cases of dengue with 1094 deaths. For 2024, a total of 1,978,372 suspected cases of dengue were reported only until the 11th epidemiological week, with 656 deaths. When comparing dengue cases reported in 2024 and 2023, there is an increase in suspected cases, with 20% more cases reported during the first 11 epidemiological weeks of 2024 than in the entire 52 epidemiological weeks of 2023. At the same time, in 2024, the Pan American Health Organization reported suspected cases in 20 Latin American countries, with 3073 cases of severe dengue and 1187 deaths. In this sense, Brazil was responsible for more than 50% of suspected cases and deaths from dengue compared to the other Latin American countries in 2024. Furthermore, there is a different ethnic racial profile for dengue in Brazil, as Black people, Mixed individuals, and Indigenous peoples were more likely to be suspected cases of dengue in 2024 compared to 2023.

Impact on Aedes aegypti Mosquitoes Exposed to Honey-Impregnated Flinders Technology Associates (FTA®) Cards.

Programs to control viruses transmitted by mosquitoes requires the implementation of surveillance tools. Over the past decade, Flinders Technology Associates (FTA®) cards, which preserve nucleic acids, have emerged as an innovating surveillance system for collecting arboviruses expectorated during mosquito sugar feeding. In this study, we evaluate the survival rates of two strain of Aedes aegypti (New Orleans (NO) and Cayenne (CAY)) in the laboratory after exposing to either honey-impregnated FTA® cards or untreated filter paper (UFP) card. Experimental exposure of mosquitoes to FTA® cards during sugar feeding significantly negatively impacted their survival, as compared to untreated filter paper. The median survival time was 2 days (95% confidence interval [CI] 1 day, 3 days) for mosquitoes exposed to FTA cards from strain NO and 3 days (95% CI 2 days, 3 days) for mosquitoes exposed to FTA cards from strain CAY. Mosquitoes exposed to UFP did not survive until the end of the experiment (4 days for strain NO and 7 days for strain CAY). Although this finding does not preclude the use of FTA® cards in surveillance, it is crucial to consider and incorporate this factor into study designs.

Executive Summary of the Spanish Guidelines for the Diagnosis and Management of Imported Febrile Illnesses from the Spanish Society of Tropical Medicine and International Health (SEMTSI), the Imported Pathology Group of the Spanish Society of Infectious Diseases and Clinical Microbiology (GEPI-SEIMC), the Spanish Society of Family and Community Medicine (SEMFYC), the Spanish Society of Primary Care Physicians (SEMERGEN) and the Spanish Society of Emergency Medicine (SEMES).

The Spanish Society of Tropical Medicine and International Health (SEMTSI), the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC), the Spanish Society of Emergency Medicine (SEMES), the Spanish Society of Primary Care Physicians (SEMERGEN) and the Spanish Society of Family and Community Medicine (SEMFYC) have prepared a consensus statement on the diagnosis and management of patients with imported febrile illnesses. Twenty authors with different backgrounds and representing different healthcare perspectives (ambulatory primary care, travel and tropical medicine specialists, emergency medicine, hospital care, microbiology and parasitology and public health), identified 39 relevant questions, which were organised in 7 thematic blocks. After a systematic review of the literature and a thoughtful discussion, the authors prepared 125 recommendations, as well as several tables and figures to be used as a consulting tool. The present executive summary shows a selection of some of the most relevant questions and recommendations included in the guidelines.

Epidemiological, virological and clinical characterization of a Dengue/Zika outbreak in the Caribbean region of Costa Rica 2017-2018.

The increase in incidence and geographical expansion of viruses transmitted by the Aedes mosquitoes, such as dengue (DENV) and zika (ZIKV) in the Americas, represents a burden for healthcare systems in tropical and subtropical regions. These and other under-detected arboviruses co-circulate in Costa Rica, adding additional complexity to their management due to their shared epidemiological behavior and similarity of symptoms in early stages. Since diagnostics of febrile illness is mostly based on clinical symptoms alone, we gathered acute-phase serum and urine from 399 samples of acute dengue-like cases from two healthcare facilities of Costa Rica, during an outbreak of arboviruses from July 2017 to May 2018, and tested them using molecular and serological methods. The analyses showed that of the clinically presumptive arbovirus cases that were reported, only 39.4% (n=153) of the samples were confirmed positive by RT-PCR to be DENV (DENV (10.3%), CHIKV (0.2%), ZIKV (27.3%), or mixed infections (1.5%). RT-PCR for other alphaviruses and flaviviruses, and PCR for Leptospira sp were negative. Furthermore, to assess flavivirus positivity in post-acute patients, the negative sera were tested against Dengue-IgM. 20% of sera were found positive, confounding even more the definitive number of cases, and emphasizing the need of several distinct diagnostic tools for accurate diagnostics. Molecular characterization of the prM and E genes from isolated viruses revealed that the American/Asian genotype of DENV-2 and the Asian lineage of ZIKV were circulating during this outbreak. Two different clades of DENV-2 American/Asian genotype were identified to co-circulate in the same region and a difference in the platelet and leukocyte count was noted between people infected with each clade, suggesting a putative distinct virulence. Our study sheds light on the necessity for healthcare strategies in managing arbovirus outbreaks, emphasizing the importance of comprehensive molecular and serological diagnostic approaches, as well as molecular characterization. This approach aids in enhancing our understanding of the clinical and epidemiological aspects of arboviral diseases during outbreaks. Our research highlights the need to strengthen training programs for health professionals and the need to increase research-based on laboratory evidence for diagnostic accuracy, guidance, development and implementation of public health interventions and epidemiological surveillance.

Urban arbovirus exposure in blood donations from an endemic area of Brazil.

BACKGROUND AND OBJECTIVES: In Brazil, urban arboviruses, such as dengue virus (DENV), Zika virus (ZIKV) and chikungunya virus (CHIKV), constitute a major public health problem, and due to their endemicity and asymptomatic cases, they pose a potential threat to blood donations. Rio de Janeiro (RJ), Brazil, has been impacted by extensive DENV epidemics over the last 30 years and, after 2015, by CHIKV and ZIKV. MATERIALS AND METHODS: Urban arboviruses DENV, ZIKV and CHIKV were investigated in blood donations (n = 778) at the State Institute of Hematology, HEMORIO (RJ) from 2019 to 2022 by serological and molecular methods. RESULTS: An overall arbovirus exposure was observed in 26.1% of the blood donations. Anti-DENV IgM was detected in 4.0% of samples and two donations were DENV NS1 positive. Positive anti-CHIKV IgM was observed in 4.7% of the donations. Co-detection of anti-CHIKV IgM and anti-DENV IgM was observed in 1.0% of donors, and CHIKV prevalence was 21.3%. All blood donations tested were negative for the DENV, ZIKV and CHIKV RNA. CONCLUSION: IgM seroprevalence to the arboviruses analyzed here is an indicator of recent infection in asymptomatic donors, showing that the population of blood donors can be a vehicle for new infections, especially during epidemic periods.

Morphological changes in eggs and embryos of Aedes aegypti (Diptera: Culicidae) exposed to predicted climatic scenarios for the year 2100 in the Central Amazon.

According to the IPCC, by the year 2100, rises in global temperature could reach up to 5 °C above current averages. On a planet-wide scale, this is one of the effects of climate changes that could have repercussions on the biological cycle of Aedes aegypti, the main arbovirus vector in urban environments and a transmitter of the arboviruses that cause dengue, Zika, chikungunya and urban yellow fever. The objective of this study was to evaluate morphological changes in Ae. aegypti eggs and embryos maintained in a climate change simulator. For this, specimens obtained from an insectarium were kept in four chambers that simulated the range of environmental scenarios predicted by the IPCC for the year 2100. The eggs obtained from each room were collected and transported to the laboratory for morphometric and morphological analysis, using confocal and scanning microscopy. Aedes aegypti eggs (n=20) were used to obtain the following variables: total width, total length, length-width ratio and diameter of the micropylar disc. Additionally, 20 embryos were used to obtain the data on head capsule length, width and length-width ratio. The data were subjected to a normality test and the means of each variable were compared using ANOVA and Tukey's post-hoc test, considering (p ≤ 0.05). A significant reduction (p < 0.05) was observed mainly in the mean lengths under the current-extreme scenario (587.5 and 553.6 µm, respectively), as well as in the widths under the current-mild scenario (171 and 158.4 µm, respectively). The length of the cephalic capsule was also affected, showing significant differences in the means under the current-intermediate scenario (189.5 and 208.5 µm, respectively), as well as in the widths between the current-intermediate scenarios (173.7 and 194.9 µm, respectively). The results suggest significant changes in the morphometry of Ae. aegypti eggs and embryos as a result of the climatic influences to which the adults were subjected, which may have an impact on vector population density and, consequently, on arbovirus dynamics in urban environments.

Underdiagnosis of arbovirus infections in returned travelers in an area with growing circulation of Aedes albopictus in North-Eastern France.

BACKGROUND: Vector-borne diseases such as malaria and arboviruses are common etiologies of post-travel fever. METHODS: After excluding malaria, we retrospectively analyzed the diagnosis of dengue virus (DENV), chikungunya virus (CHIKV), and zika virus (ZIKV) infections following recent travel by patients treated at the Strasbourg University Hospital between 2014 and 2023. Available serums (n = 35) sampled in 2023 were retrospectively tested for DENV, CHIKV, and ZIKV infections. RESULTS: Our results showed that 78% of the 915 malaria-negative patients without changes over the course of ten years had not undergone arbovirus infection testing. Retrospective testing revealed missing arbovirus infections: two DENV infections and one CHIKV infection, representing 8.6% (3/35) of patients for whom no mandatory declaration or vector control could be undertaken. CONCLUSION: Our results highlight the need for early case detection, particularly in the context of the upcoming 2024 Olympic Games.

Crimean-Congo Hemorrhagic Fever Virus Kinetics in Serum, Saliva, and Urine, Iran, 2018.

Little is known about using noninvasive samples for diagnosing Crimean-Congo hemorrhagic fever (CCHF). We investigated detection of CCHF virus in serum, saliva, and urine samples. Our results indicate that serum is the best sample type for CCHF diagnosis; saliva can be used for noninvasive sampling.

Aptamer-associated colorimetric reverse transcription loop-mediated isothermal amplification assay for detection of dengue virus.

Current diagnostic methods for dengue, such as serological tests, have limitations in terms of cross-reactivity with other viruses. To address this issue, we explored the potential of combining the loop-mediated isothermal amplification (LAMP) technique with the affinity of aptamers to develop point-of-care testing. In this study, we utilized 60 serum samples. An aptamer capable of binding to the dengue virus was employed as a platform for capturing genetic material, and its performance was compared to a commercial kit. Dengue virus was detected through RT-PCR and colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP), allowing visual observation of the results without the need for equipment. In the context of the aptamer LAMP assay, our analysis revealed the detection of the dengue virus in 38 out of 60 samples, with 95% sensitivity and 100% specificity compared to RT-PCR and/or APTA-RT-PCR. Importantly, we observed no cross-reaction when assessing samples positive for the zika virus, underscoring the assay's selectivity. This innovative aptameric capture of the viral RNA in combination with the RT-LAMP (APTA-RT-LAMP) method has the potential to offer valuable molecular insights into neglected infectious diseases in a simpler and faster manner. IMPORTANCE: Dengue is a neglected tropical disease of significant epidemiological importance in tropical and subtropical countries. Current diagnostics for this infection present challenges, such as cross-reactivity in serological tests. Finding ways to enhance the diagnosis of this disease is crucial, given the absence of specific treatments. An accurate, simple, and effective diagnosis contributes to the improved management of infected individuals. In this context, our work combines molecular biology techniques, such as isothermal loop amplification, with aptamers to detect the dengue virus in biological samples. Our method produces colorimetric results based on a color change, with outcomes available in less than 2 hours. Moreover, it requires simpler equipment compared to molecular PCR tests.

Análise bibliométrica da produção científica global em chikungunya: perspectiva da Vigilância em Saúde / Bibliometric analysis of global scientific production on chikungunya: Health Surveillance perspective / Análisis bibliométrico de la producción científica mundial sobre chikungunya: una perspectiva de Vigilancia Sanitária

Chikungunya, arbovirose que provoca febre e artralgia debilitante, demonstra potencial crônico e incapacitante por longos períodos, não havendo vacinas ou terapias específicas. Recentemente, a doença evoluiu da condição negligenciada para uma ameaça à saúde pública, impactando milhões de pessoas em regiões tropicais e subtropicais. Este estudo analisa a produção do conhecimento sobre a Chikungunya, na perspectiva da Vigilância em Saúde. Trata-se de metodologia exploratória-descritiva, com análise bibliométrica. Realizou-se a coleta nas bases Scopus e Web of Science para artigos de 2008 a 2022. A análise revelou uma série temporal de produção destacando a contribuição de Estados Unidos, Brasil e França. Identificaram-se como áreas mais produtivas: doenças transmissíveis, medicina tropical e parasitologia, fundamentais à Vigilância em Saúde e ao planejamento de políticas públicas. Quanto aos pesquisadores, Weaver, Scott C., Failloux, Anna-Bella e De Lamballerie, Xavier foram relevantes no cenário global, indicando a importância da colaboração e da abordagem interdisciplinar.

Chikungunya, an arbovirus that causes fever and debilitating arthralgia, has potential to be chronic and incapacitating for long periods, and there are no vaccines or therapies available for it. Recently, the disease has evolved from a neglected condition to public health threat, impacting millions in tropical and subtropical regions. This study analyzed the knowledge production about chikungunya, from the perspective of Health Surveillance, using an exploratory-descriptive methodology and bibliometric analysis. Articles from 2008 to 2022 were collected from Scopus and Web of Science databases. The analysis showed a production time series, highlighting the contribution of United States, Brazil, and France. The most productive areas were identified as: communicable diseases, tropical medicine and parasitology, which are fundamental to Health Surveillance and public policy planning. Regarding researchers, Weaver, Scott C., Failloux, Anna-Bella, and De Lamballerie, Xavier were relevant in the global scenario, indicating the importance of collaboration and of the interdisciplinary approach.

El chikungunya, arbovirus que causa fiebre y artralgia debilitante, tiene potencial de ser crónico e incapacitante por largos periodos, todavía no hay vacunas ni terapias. Recientemente, la enfermedad ha pasado de afección desatendida a amenaza para la salud pública, afectando a millones en regiones tropicales y subtropicales. Este estudio analiza la producción de conocimientos sobre chikungunya, desde la perspectiva de la Vigilancia Sanitaria. Se utilizó una metodología exploratoria-descriptiva con análisis bibliométrico. Se recompilaron artículos de Scopus y Web of Science, 2008-2022. El análisis reveló una serie temporal de producción, destacando la contribución de EEUU, Brasil y Francia. Se identificaron como áreas más productivas: enfermedades transmisibles, medicina tropical y parasitología, fundamentales para la Vigilancia Sanitaria y la planificación de políticas públicas. En cuanto a los investigadores, Weaver, Scott C., Failloux, Anna-Bella y De Lamballeire, Xavier fueron relevantes en el escenario global, indicando la importancia de la colaboración y del enfoque interdisciplinario.

Utilization of machine learning for dengue case screening.

Dengue causes approximately 10.000 deaths and 100 million symptomatic infections annually worldwide, making it a significant public health concern. To address this, artificial intelligence tools like machine learning can play a crucial role in developing more effective strategies for control, diagnosis, and treatment. This study identifies relevant variables for the screening of dengue cases through machine learning models and evaluates the accuracy of the models. Data from reported dengue cases in the states of Rio de Janeiro and Minas Gerais for the years 2016 and 2019 were obtained through the National Notifiable Diseases Surveillance System (SINAN). The mutual information technique was used to assess which variables were most related to laboratory-confirmed dengue cases. Next, a random selection of 10,000 confirmed cases and 10,000 discarded cases was performed, and the dataset was divided into training (70%) and testing (30%). Machine learning models were then tested to classify the cases. It was found that the logistic regression model with 10 variables (gender, age, fever, myalgia, headache, vomiting, nausea, back pain, rash, retro-orbital pain) and the Decision Tree and Multilayer Perceptron (MLP) models achieved the best results in decision metrics, with an accuracy of 98%. Therefore, a tree-based model would be suitable for building an application and implementing it on smartphones. This resource would be available to healthcare professionals such as doctors and nurses.

Exploring the urban arbovirus landscape in Rio de Janeiro, Brazil: transmission dynamics and patterns of disease spread.

Background: This study focuses on urban arboviruses, specifically dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV), which pose a significant public health challenge in Rio de Janeiro state, Southeast Brazil. In our research, we highlight critical findings on the transmission dynamics of these arboviruses in Rio de Janeiro, identifying distinct patterns of disease spread. Methods: By combining genomic data with case reports from the Brazilian Ministry of Health, we have analysed the phylogenetics, prevalence and spatial distribution of these endemic viruses within the state. Findings: Our results revealed sustained DENV transmission primarily in the northern part of the state, a significant ZIKV epidemic in 2016 affecting all mesoregions, and two major CHIKV outbreaks in 2018 and 2019, predominantly impacting the northern and southern areas. Our analysis suggests an inverse relationship between arboviral case incidence and urban density, with less populous regions experiencing higher transmission rates, potentially attributed to a complex interplay of factors such as the efficacy of vector control measures, environmental conditions, local immunity levels, and human mobility. Furthermore, our investigation unveiled distinct age and gender trends among affected individuals. Notably, dengue cases were predominantly observed in young adults aged 32, while chikungunya cases were more prevalent among individuals over 41. In contrast, cases of ZIKV were concentrated around the 33-year age group. Intriguingly, females accounted for nearly 60% of the cases, suggesting a potential gender-based difference in infection rates. Interpretation: Our findings underscore the complexity of arbovirus transmission and the need for interventions tailored to different geographical mesoregions. Enhanced surveillance and genomic sequencing will be essential for a deeper, more nuanced understanding of regional arbovirus dynamics. Identifying potential blind spots within the state will be pivotal for developing and implementing more effective public health strategies, specifically designed to address the unique challenges posed by these viruses throughout the state. Funding: This study was supported by the National Institutes of Health USA grant U01 AI151698 for the United World Arbovirus Research Network (UWARN) and the CRP-ICGEB RESEARCH GRANT 2020 Project CRP/BRA20-03.

A meta-analysis of Chikungunya virus in neurological disorders.

Chikungunya disease typically presents with the fever-arthralgia-rash symptom triad. However, an increase in the number of atypical clinical manifestations, particularly neurological disorders, has occurred. The current evidence regarding the pooled prevalence of Chikungunya virus (CHIKV)-associated neurological cases (CANCs) suspected of having an arboviral aetiology is not well-understood. Therefore, this meta-analysis included 19 studies (n = 7319 patients) and aimed to determine the pooled rate of exposure to CANC. The pooled positivity rate of CANC was 12 % (95 % CI: 6-19), and Brazil was overrepresented (11/19). These estimations varied between 3 and 14 % based on the diagnostic method (real-time PCR vs. ELISA-IgM) and biological samples (cerebrospinal fluid or blood specimens) used for detection of CHIKV. Regarding the frequency of CHIKV in neurological clinical subgroups, the rates were higher among patients with myelitis (27 %), acute disseminated encephalomyelitis (27 %), Guillain-Barré syndrome (15 %), encephalitis (12 %), and meningoencephalitis (7 %). Our analysis highlights the significant burden of CANC. However, the data must be interpreted with caution due to the heterogeneity of the results, which may be related to the location of the studies covering endemic periods and/or outbreaks of CHIKV. Current surveillance resources should also focus on better characterizing the epidemiology of CHIKV infection in neurological disorders. Additionally, future studies should investigate the interactions between CHIKV and neurological diseases with the aim of gaining deeper insight into the mechanisms underlying the cause-and-effect relationship between these two phenomena.

Mitochondrial and microbial diversity of the invasive mosquito vector species Culex tritaeniorhynchus across its extensive inter-continental geographic range.

Background: Culex (Cx.) tritaeniorhynchus is an invasive mosquito species with an extensive and expanding inter-continental distribution, currently reported across Asia, Africa, the Middle East, Europe and now Australia. It is an important vector of medical and veterinary pathogens which cause significant morbidity and mortality in human and animal populations. Across regions endemic for Japanese encephalitis virus (JEV), Cx. tritaeniorhynchus is considered the major vector and has also been shown to contribute to the transmission of several other zoonotic arboviruses including Rift Valley fever virus (RVFV) and West Nile virus (WNV). Methods: In this study, we used laboratory vector competence experiments to determine if Cx. tritaeniorhynchus from a Southern European population were competent JEV vectors. We also obtained samples from multiple geographically dispersed Cx. tritaeniorhynchus populations from countries within Europe, Africa, Eurasia and Asia to perform phylogenetic analysis to measure the level of mitochondrial divergence using the cytochrome oxidase subunit 1 ( CO1) gene. We also undertook bacterial 16S rRNA gene amplicon sequencing to determine microbial diversity and used multi-locus sequence typing (MLST) to determine any evidence for the presence of strains of the naturally occurring endosymbiotic bacterium Wolbachia. Results: Cx. tritaeniorhynchus from a Greek population were shown be be competent vectors of JEV with high levels of virus present in saliva. We found a signficant level of mitochondrial genetic diversity using the mosquito CO1 gene between geographically dispersed populations. Furthermore, we report diverse microbiomes identified by 16S rRNA gene amplicon sequencing within and between geographical populations. Evidence for the detection of the endosymbiotic bacteria Wolbachia was confirmed using Wolbachia-specific PCR and MLST. Conclusions: This study enhances our understanding of the diversity of Cx. tritaeniorhynchus and the associated microbiome across its inter-continental range and highlights the need for greater surveillance of this invasive vector species in Europe.

The mosquito species Culex (Cx.) tritaeniorhynchus is expanding its range and is now present in over 50 countries across Asia, Africa, the Middle East, Europe and now Australasia. It can transmit human and animal pathogens, resulting in significant morbidity and mortality. This species transmits Japanese encephalitis virus in endemic areas of Asia, and it has also been shown to contribute to the transmission of several other viruses that can infect humans, including Rift Valley fever virus and West Nile virus. In this study, we firstly undertook some lab experiments to show that Cx. tritaeniorhynchus from a Southern European population are competent vectors of Japanese encephalitis virus. We also obtained field mosquitoes from countries within Europe, Africa, Eurasia and Asia and used phylogenetic analysis to demonstrate a high level of mitochondrial divergence within and between populations. In addition, we analysed the bacteria present within mosquitoes and found a high level of microbial diversity. Finally, we present evidence for the presence of Wolbachia endosymbiotic bacteria in some populations of this mosquito species. This study highlights the need for greater surveillance of this invasive vector species ­ particularly in Europe.

Neuroinvasion of emerging and re-emerging arboviruses: A scoping review.

Background: Arboviruses are RNA viruses and some have the potential to cause neuroinvasive disease and are a growing threat to global health. Objectives: Our objective is to identify and map all aspects of arbovirus neuroinvasive disease, clarify key concepts, and identify gaps within our knowledge with appropriate future directions related to the improvement of global health. Methods: Sources of Evidence: A scoping review of the literature was conducted using PubMed, Scopus, ScienceDirect, and Hinari. Eligibility Criteria: Original data including epidemiology, risk factors, neurological manifestations, neuro-diagnostics, management, and preventive measures related to neuroinvasive arbovirus infections was obtained. Sources of evidence not reporting on original data, non-English, and not in peer-reviewed journals were removed. Charting Methods: An initial pilot sample of 30 abstracts were reviewed by all authors and a Cohen's kappa of κ = 0.81 (near-perfect agreement) was obtained. Records were manually reviewed by two authors using the Rayyan QCRI software. Results: A total of 171 records were included. A wide array of neurological manifestations can occur most frequently, including parkinsonism, encephalitis/encephalopathy, meningitis, flaccid myelitis, and Guillain-Barré syndrome. Magnetic resonance imaging of the brain often reveals subcortical lesions, sometimes with diffusion restriction consistent with acute ischemia. Vertical transmission of arbovirus is most often secondary to the Zika virus. Neurological manifestations of congenital Zika syndrome, include microcephaly, failure to thrive, intellectual disability, and seizures. Cerebrospinal fluid analysis often shows lymphocytic pleocytosis, elevated albumin, and protein consistent with blood-brain barrier dysfunction. Conclusions: Arbovirus infection with neurological manifestations leads to increased morbidity and mortality. Risk factors for disease include living and traveling in an arbovirus endemic zone, age, pregnancy, and immunosuppressed status. The management of neuroinvasive arbovirus disease is largely supportive and focuses on specific neurological complications. There is a need for therapeutics and currently, management is based on disease prevention and limiting zoonosis.

First Record of the Tribe Orthopodomyiini in the Yucatan Peninsula.

Updating the mosquito fauna occurring in a specific area is crucial, given that certain species serve as vectors capable of transmitting zoonotic arboviruses. This scientific note presents the first records of mosquitoes of the tribe Orthopodomyiini in the Yucatan Peninsula. Immature mosquitoes were collected on 2 occasions inside a large tree hole in Felipe Carrillo Puerto, Quintana Roo, Mexico. Thirteen adult specimens, reared from the immatures, were obtained and identified as Orthopodomyia kummi based on external characteristics of females and males. This species has been recorded in Panama, Costa Rica, El Salvador, Guatemala, Mexico, and marginally in the United States, but its presence in the Yucatan Peninsula had gone unnoticed until now. The knowledge about mosquitoes of the genus Orthopodomyia is limited, and their epidemiological importance remains uncertain. Therefore, further studies could provide insights into the ecological and infection dynamics associated with this species.

In vivo rescue of arboviruses directly from subgenomic DNA fragments.

Reverse genetic systems are mainly used to rescue recombinant viral strains in cell culture. These tools have also been used to generate, by inoculating infectious clones, viral strains directly in living animals. We previously developed the "Infectious Subgenomic Amplicons" (ISA) method, which enables the rescue of single-stranded positive sense RNA viruses in vitro by transfecting overlapping subgenomic DNA fragments. Here, we provide proof-of-concept for direct in vivo generation of infectious particles following the inoculation of subgenomic amplicons. First, we rescued a strain of tick-borne encephalitis virus in mice to transpose the ISA method in vivo. Subgenomic DNA fragments were amplified using a 3-fragment reverse genetics system and inoculated intramuscularly. Almost all animals were infected when quantities of DNA inoculated were at least 20 µg. We then optimized our procedure in order to increase the animal infection rate. This was achieved by adding an electroporation step and/or using a simplified 2- fragment reverse genetics system. Under optimal conditions, a large majority of animals were infected with doses of 20 ng of DNA. Finally, we demonstrated the versatility of this method by applying it to Japanese encephalitis and Chikungunya viruses. This method provides an efficient strategy for in vivo rescue of arboviruses. Furthermore, in the context of the development of DNA-launched live attenuated vaccines, this new approach may facilitate the generation of attenuated strains in vivo. It also enables to deliver a substance free of any vector DNA, which seems to be an important criterion for the development of human vaccines.

Aedes albopictus is a competent vector of five arboviruses affecting human health, greater Paris, France, 2023.

Aedes albopictus collected in 2023 in the greater Paris area (Île-de-France) were experimentally able to transmit five arboviruses: West Nile virus from 3 days post-infection (dpi), chikungunya virus and Usutu virus from 7 dpi, dengue virus and Zika virus from 21 dpi. Given the growing number of imported dengue cases reported in early 2024 in France, surveillance of Ae. albopictus should be reinforced during the Paris Olympic Games in July, when many international visitors including from endemic countries are expected.

Advancing arbovirus diagnosis in Brazil: strengthening diagnostic strategies and public health data collection.

BACKGROUND: The last five decades have seen a surge in viral outbreaks, particularly in tropical and subtropical regions like Brazil, where endemic arboviruses such as Dengue (DENV), Zika (ZIKV), and Chikungunya (CHIKV) pose significant threats. However, current diagnostic strategies exhibit limitations, leading to gaps in infection screening, arbovirus differential diagnoses, DENV serotyping, and life-long infection tracking. This deficiency impedes critical information availability regarding an individual's current infection and past infection history, disease risk assessment, vaccination needs, and policy formulation. Additionally, the availability of point-of-care diagnostics and knowledge regarding immune profiles at the time of infection are crucial considerations. OBJECTIVES: This review underscores the urgent need to strengthen diagnostic methods for arboviruses in Brazil and emphasizes the importance of data collection to inform public health policies for improved diagnostics, surveillance, and policy formulation. METHODS: We evaluated the diagnostic landscape for arboviral infections in Brazil, focusing on tailored, validated methods. We assessed diagnostic methods available for sensitivity and specificity metrics in the context of Brazil. RESULTS: Our review identifies high-sensitivity, high-specificity diagnostic methods for arboviruses and co-infections. Grifols transcription-mediated amplification assays are recommended for DENV, CHIKV, and ZIKV screening, while IgG/IgM ELISA assays outperform Rapid Diagnostic Tests (RDTs). The Triplex real-time RT-PCR assay is recommended for molecular screening due to its sensitivity and specificity. CONCLUSION: Enhanced diagnostic methods, on-going screening, and tracking are urgently needed in Brazil to capture the complex landscape of arboviral infections in the country. Recommendations include nationwide arbovirus differential diagnosis for DENV, ZIKV, and CHIKV, along with increased DENV serotyping, and lifelong infection tracking to combat enduring viral threats and reduce severe presentations.