How human papillomavirus (HPV) vaccines elicit robust and enduring neutralizing antibody (nAb) responses is unknown, yet such information is valuable to vaccinology. In addition to the major capsid protein, L1, the 4-valent HPV vaccine reportedly also contains recombinant L1 DNA. Nucleic acids in vaccines enhance antibody responses and are employed as adjuvants, but whether L1 DNA enhances HPV vaccine antibody responses is understudied. We tested whether 9-valent HPV (9vHPV) vaccine lots contained L1 DNA and if peak or long-term 9vHPV vaccine-elicited nAb responses were dependent on DNA sensors Toll-like receptor 9 (TLR9), AIM2, or cGAS or on STING or MyD88, the adaptors for cGAS and TLR9. To quantify L1 DNA, we extracted total nucleic acid per 9vHPV dose and applied quantitative PCR to amplify HPV6/11/16/18/31/33/45/52/58 L1 sequences. Wild-type mice (C57BL/6J) or mice deficient in DNA sensing pathways (TLR9-/-, AIM2-/-, cGAS-/-, STINGgt/gt, MyD88-/-) were administered 9vHPV vaccine or equivalent adjuvant only at 0, 4, and 12 weeks. We measured serum HPV16 and HPV18 nAb titers at 16, 26, 40, and 53-56 weeks and HPV16- and HPV18-specific bone marrow plasma cell responses at 53-56 weeks. We detected 5-44 and 104-351 copies of HPV6 and HPV18 L1 DNA per 9vHPV dose, but no other types (n = 3 lots). We found no consistent difference in geometric mean nAb titers between mice strains that received 9vHPV at any time point tested or in median HPV-specific plasma cell frequencies. Thus, we conclude the magnitude and durability of nAb responses to 9vHPV vaccination do not depend on DNA sensing pathways.IMPORTANCEHow the highly effective human papillomavirus (HPV) vaccines elicit strong and long-lasting antibody responses is unknown, yet such knowledge is valuable to vaccine development. The HPV vaccines are comprised of virus-like particles assembled from the HPV major capsid protein, L1; an adjuvant; and reportedly also residual recombinant L1 DNA as a result of the manufacturing process. We tested whether the currently available 9-valent HPV (9vHPV) vaccine contained L1 DNA and if 9vHPV vaccine-elicited antibody responses were dependent on DNA sensing pathways of the innate immune system in vivo. We confirmed separate lots of the 9vHPV vaccine contained low copy numbers of L1 DNA, but found 9vHPV vaccine-generated antibody responses were not dependent on DNA sensing. This study has thus refined our understanding of potential mechanisms underlying the strong and long-lasting antibody responses to HPV vaccination.

The emergence of COVID-19 facilitated the widespread use, regulatory awareness, and societal familiarity of lateral flow assays (LFA) for self-diagnosis. The increased frequency in the use of LFA’s as well as their increased use by the general population amplified some of the major drawbacks of these tests, namely, the reliance on user interpretation of the test result, the qualitative nature of the result itself, and the lower sensitivity and specificity compared to traditional polymerase chain reaction (PCR)-based diagnostic testing. Furthermore, the results of these tests and the interpretation of them by lay users were increasingly used as a public health tool by authorities, where recommendations were made on how members of the public should act in response to self-interpretation of test results. Our aims were to develop an objective image analysis algorithm that can (1) interpret the result of a popular COVID-19 LFA Rapid Antigen Test (RAT) and accurately estimate viral load, and (2) predict how long after a positive test a user will test negative on the RAT, indicating loss of infectiousness and the ability to be around others without fear of spreading infection. Indeed, we found that we could estimate viral load to within a RMSE of 1.44 Cycle Threshold (CT) of gold standard PCR testing, and that we can predict the result status of a user upon a follow up visit at least 5 days post-symptom onset with a sensitivity of 99

Background Nipah virus (NiV) is a highly pathogenic, zoonotic paramyxovirus with significant public health implications due to high associated mortality and potential for human-to-human transmission. Current diagnostic testing options for NiV are limited and require extensive laboratory infrastructure. Objective Develop a field-deployable testing workflow for timely NiV detection. Study design A NiV real-time RT-PCR (rRT-PCR) was designed for a highly conserved region of the nucleocapsid gene and tested with RNA from Bangladesh and Malaysia NiV strains. The NiV rRT-PCR was evaluated on Rotor-Gene Q and Palm PCR S1e thermocyclers following instrument free RNA extraction (Extract & Store). Results Initial analytical evaluation, on a Rotor-Gene Q, demonstrated dynamic amplification and a limit of detection (LoD) of 3.7-4.2 copies/µL without amplification of related paramyxoviruses. The assay was adapted for the portable, battery-powered, self-contained Palm PCR S1e thermocycler, and exhibited linear detection with a LoD of 30.7 copies/µL. RNA extraction from contrived whole blood and pharyngeal swabs using the Extract & Store workflow yielded comparable results to automated extraction on a KingFisher Apex instrument. The entire assay, including extracted and stabilized RNA controls from BSL-1 strains, was successfully transferred to Aga Khan University with ambient temperature shipping and yielded similar performance. Conclusions The combination of Extract & Store and the Palm PCR S1e device offers a viable solution for field-based molecular detection of NiV. While limitations were noted for reaction setup on the Palm PCR, this presents a flexible and accessible workflow for rapid, portable detection of high-consequence pathogens in resource-constrained settings.

BACKGROUND:Recent global epidemics and pandemics have highlighted the need for adequate diagnostic capacities to respond to emerging threats. Strengthening operational capacities can lead to improved preparation; investing in emergency response infrastructure may diminish progression of outbreaks and prevent escalation. METHODS:An email-linked survey was sent January-March-2025 to two global infectious disease surveillance networks' (GeoSentinel and TropNet) participating clinics assessing capacity for Oropouche virus (OROV) diagnostic testing. Data on testing capacity for other arboviruses was also requested. RESULTS:Responses were obtained for 58 GeoSentinel/TropNet sites (58/104, 55.8%): 50% responding sites (29/58) had OROV diagnostic testing available at a public reference laboratory; 23 sites (40%) had capacity for OROV RNA detection; 10 sites (17%) had OROV serological testing onsite. Estimated median turn-around times from phlebotomy-molecular OROV results were 4 days onsite and 6 days at the reference laboratory. Viral culture was available onsite at 9% of sites (at the reference laboratory for 26%). Molecular diagnostics availability for other arboviruses onsite showed >50% could test for at least one other arbovirus: dengue virus (31/58, 53%), chikungunya virus (29/58, 50%), West Nile virus (22/58, 38%) and yellow fever virus (12/58, 21%). Onsite serologic testing for non-OROV arboviruses was available at 39/58 sites (67%). CONCLUSIONS:The survey identified potential weaknesses in novel pathogen preparedness with respect to diagnostic capacity for OROV testing at specialized travel medicine clinics participating in two international networks. These findings provide an opportunity to implement measures to improve communication and planning and to strengthen diagnostic infrastructure prior to future outbreaks.

Molecular tests offer a powerful approach to diagnosing numerous diseases and, due to advances in lab-on-a-chip technologies, have the potential to be used in the field and at the bedside. However, there is still a need for portable technologies that help isolate nucleic acids from their associated sample for testing. Previously, our groups developed an RNA extraction and stabilization (RNAES) protocol that utilized novel shelf stable reagents to extract, isolate, and store nucleic acids on membranes at ambient temperature. Building on these results, we have created RNAES kits with reductionist, 3D-printed, lab-on-a-chip style parts that enable field isolation of RNA without external equipment or power. Our kits feature three components, a lysis sphere, a capillary-wicking based cartridge for RNA isolation, as well as a drying & storage tube that stabilizes RNA for ambient temperature shipping. Using our kit, a user can process a 25 μL sample of serum or plasma within 15 minutes, including a 10-minute incubation time. Noting that hepatitis C virus (HCV) can be cured when detected, we use our RNAES kit in combination with a new real-time RT-PCR to detect HCV. Our data on 98 clinical samples and four contrived whole blood samples show comparable RNA detection and quantified viral loads following extraction with our RNAES kit or a commercial system. Our approach also provided the additional benefit of stabilizing viral RNA for storage at ambient temperatures as lysate, for up to 7 days, or on dried membranes in storage tubes, for at least 30 days.

Importance:Diarrheal disease causes substantial morbidity in international travelers. Knowledge of antibiotic resistance of causative pathogens helps guide empiric treatment decisions. Objective:To characterize antimicrobial nonsusceptibility patterns of culture-confirmed Campylobacter species, nontyphoidal Salmonella (NTS) species, Shigella species, and diarrheagenic Escherichia coli isolated from international travelers with diarrhea identified by the GeoSentinel Surveillance Network. Design, Setting, and Participants:This is a retrospective cross-sectional analysis of antimicrobial susceptibility data for 4 major pathogens causing travel-associated diarrhea, reported from April 14, 2015, to December 19, 2022, at 58 of 71 international GeoSentinel sites. Participants included a convenience sample of international travelers with acute diarrhea seen during or after travel presenting at the GeoSentinel sites. Main Outcomes and Measures:The main outcomes were demographics, clinical characteristics, and antimicrobial susceptibility profiles of patients with culture-confirmed Campylobacter species, NTS species, Shigella species, and diarrheagenic E coli species. Routinely collected antimicrobial susceptibility data with intermediate susceptibility and resistant were defined as nonsusceptible. The antimicrobial susceptibility test results were described as numbers and percentages, and binomial 95% CIs were calculated for the proportions. Results:Of 859 total cases, the median (IQR) age was 30 (23-43) years, and 440 travelers (51%) were male. Among Campylobacter isolates, nonsusceptibility to fluoroquinolones was found in 206 of 274 isolates (75%; 95% CI, 70%-80%), and nonsusceptibility to macrolides was found in 30 of 255 isolates (12%; 95% CI, 8%-16%) and was highest in travelers to South Central Asia (45 of 51 isolates; 88%; 95% CI, 76%-96%). Among NTS species, 96 of 302 isolates (32%; 95% CI, 27%-37%) were nonsusceptible to fluoroquinolones, 18 of 111 isolates (16%; 95% CI, 10%-24%) were nonsusceptible to macrolides, and 15 of 273 isolates (5%; 95% CI, 3%-9%) were nonsusceptible to third-generation cephalosporins. For Shigella species, 44 of 196 isolates (22%; 95% CI, 17%-29%) were nonsusceptible to fluoroquinolones, and 36 of 103 isolates (35%; 95% CI, 26%-45%) were nonsusceptible to macrolides. For E coli, fluoroquinolone nonsusceptibility was 18% (12 of 66 isolates; 95% CI, 10%-30%). Of note, 19 of 24 Shigella isolates (79%; 95% CI, 58%-93%) were nonsusceptible to fluoroquinolones among travelers to South Central Asia, and 29 of 37 Shigella isolates (78%; 95% CI, 62%-90%) were nonsusceptible to macrolides among travelers to South America. Conclusions and Relevance:In this cross-sectional study of travelers' diarrhea antimicrobial resistance patterns, there was marked variability of nonsusceptibility to 2 major classes of antibiotics commonly used for treating travelers' diarrhea among global regions. Antimicrobial susceptibility from culture should be obtained when possible, including after pathogen detection by culture-independent methods. These findings may help inform strategies for self-treatment and clinician management of travelers' diarrhea.

BACKGROUND:Prenatal Zika virus (ZIKV) infection leads to microcephaly and adverse neurodevelopment. The effects of postnatal ZIKV infection on the developing brain are unknown. We assessed the neurodevelopmental outcomes of children exposed postnatally during the ZIKV epidemic. METHODS:A prospective study enrolled infants 0-3 months of age and their mothers, and children 1.5-3.5 years of age in rural Guatemala from 2017 and were followed for 12 months until 2019. Neurodevelopment was evaluated using the Mullen Scales of Early Learning (MSEL). ZIKV and dengue virus (DENV) infections were identified by polymerase chain reaction (PCR) using active surveillance. Serological analyses, stratified by age group flavivirus serostatus at enrollment, were conducted using a focus reduction neutralization test. RESULTS:Of 1371 enrolled participants, 1187 (86.6%) completed the study. No PCR-confirmed ZIKV infections were identified during the study period. One-third of 1.5-3.5-year-old children were ZIKV-seropositive at enrollment (likely postnatal infection). Twenty participants (5.8%) tested positive for DENV by PCR (11 infants, 5 children and 4 mothers); 15 (75%) were DENV-3 infections and 5 were DENV-2. The incidence of DENV infection in infants was 2.6%. No significant differences in MSEL scores were found between infants born seropositive versus seronegative for ZIKV or DENV. DENV seropositivity at enrollment in 1.5-5-year-old children was associated with lower MSEL scores for fine motor, visual reception and language, and microcephaly at 12 months versus seronegative children (all P < 0.05). CONCLUSIONS:Postnatal ZIKV infection in children from rural Guatemala was not associated with worse neurodevelopmental outcomes. DENV seropositivity was associated with a higher risk of microcephaly in infants and worse neurodevelopmental outcomes in children.

BackgroundPakistan has been an endemic country for dengue virus since 1994, with a significant increase in cases reported in 2022 largely due to heavy rainfall and flooding. All four serotypes of the dengue virus (DENV) are present in Pakistan, with DENV 1 and DENV 2 being the most prevalent. The current study aims to explore the clinical presentations and features of dengue fever in a tertiary care hospital.MethodologyWe enrolled and studied 349 cases of suspected and confirmed dengue presenting for care at the Aga Khan University Hospital in Karachi between June 2021 and November 2023. Collected data on cases included clinical symptoms and laboratory results including qRT-PCR and serotype characterization.FindingsThe majority of subjects enrolled (75%) had mild disease without warning signs, while 11% exhibited warning signs, 1.4% had severe dengue, and 12.6% had no dengue diagnosis. Patients with severe dengue (SD) had significantly higher levels of liver enzymes (AST and ALT) compared to those with non-severe dengue (NSD) (AST; p = 0.024 and ALT; p = 0.047). Additionally, a higher grade of thrombocytopenia was significantly associated with hospitalization (p = 0.0008), and prolonged illness (p = 0.03). Both Platelet (p < 0.0001) and WBC counts (p = 0.001) were significantly lower in dengue PCR-positive patients in comparison to Dengue PCR-negative. Among those tested for dengue serotypes, DENV 1 (34%) and DENV 2 (45%) emerged as the predominant serotypes, with mixed infections accounting for 17%. The sensitivity of q-RT PCR was found to be 87.25% and the specificity of 68.35%. qRT-PCR detected 43.5% of cases with viral fever initially screened negative by IgM or NS1.ConclusionThe epidemiology of dengue fever during a widespread outbreak in 2022 showed a predominance of DENV 1 and DENV 2 serotypes with milder phenotype of viral illness. Screening with rapid tests requires further confirmation by molecular assay in cases with dengue and dengue-like illness. The sensitivity of q-RT PCR using gold standard.

Genomic epidemiology offers insight into the transmission and evolution of respiratory viruses. We used metagenomic sequencing from negative SARS-CoV-2 rapid antigen tests to identify a wide range of respiratory viruses and generate full genome sequences. This process offers a streamlined mechanism for broad respiratory virus genomic surveillance.

Dengue virus (DENV) is a significant public health concern in Colombia, with increased transmission of DENV type 1 (DENV-1) in the departments of Risaralda and Valle del Cauca in the Central-West region of the country following a large outbreak in 2019. However, little is known about the source, genetic diversity, and evolution of circulating viruses. We obtained serum samples from individuals with acute DENV infection and analysed DENV-1 genetic diversity, phylodynamics, and phylogeography. We found that most viruses belonged to DENV-1 genotype V, and phylogenetic analysis revealed three distinct clades, each of which was most closely related to viruses from neighbouring departments of Colombia sampled over the last 5-10 years. Thus, the 2019 outbreak and subsequent DENV-1 circulation was not due to the introduction of a new lineage to the country but rather reflected local DENV-1 V dispersion and evolution. We identified amino acid positions under positive selection in structural proteins and NS1, which may have a role in immune evasion and pathogenesis. Overall, our analysis of DENV-1 V diversity, evolution, and spread within Colombia highlights the important role of genomic surveillance in understanding virus dynamics during endemic circulation and outbreaks.

While SARS-CoV-2 vaccines have shown strong efficacy, the continued emergence of new viral variants raises concerns about the ongoing and future public health impact of COVID-19, especially in locations with suboptimal vaccination uptake. We investigated viral and host factors, including vaccination status, that were associated with SARS-CoV-2 disease severity in a setting with low vaccination rates. We analyzed clinical and demographic data from 1,957 individuals in the state of Georgia, USA, coupled with viral genome sequencing from 1,185 samples. We found no specific mutations associated with disease severity. Compared to those who were unvaccinated, vaccinated individuals experienced less severe SARS-CoV-2 disease, and the effect was similar for both variants. Vaccination within the prior 3-9 months was associated with decreased odds of moderate disease, severe disease, and death. Older age and underlying health conditions, especially immunosuppression and renal disease, were associated with increased disease severity. Overall, this study provides insights into the impact of vaccination status, variants/mutations, and clinical factors on disease severity in SARS-CoV-2 infection when vaccination rates are low. Understanding these associations will help refine and reinforce messaging around the crucial importance of vaccination in mitigating the severity of SARS-CoV-2 disease.

Historically, DENV-4 has been rarely associated with epidemics and has been less well-studied than DENV-1 to -3. Epidemic dengue struck several South and Central American countries in 2022, with Nicaragua reporting the highest incidence. In an acute febrile illness (AFI) cohort enrolled from June to September 2022, 58 (34%) of 172 patients had PCR-confirmed dengue, of which 46 (79%) were serotyped as DENV-4. In this cohort, acute dengue, as a proportion of AFI, increased from 8% in June to a peak of 58% in August. Genome sequencing and phylogenetic analysis identified a lineage of DENV-4 Genotype IIb (GIIb) with six amino acid substitutions on the surface-exposed regions of the envelope (E) protein as compared to a reference sequence from 2005. Indeed, two of these mutations appear to be novel and located at G172E or near N174K, an antigenic epitope on domain I. Most (90%, 43/48) DENV-4 patients had pre-existing DENV IgG (secondary dengue), at the acute phase. Secondary dengue was associated with the male sex (prevalence ratio (PR)), 6.88) and being younger than 11 years of age (PR, 8.38). Further analysis showed no association between past Zika exposure and DENV-4 acute illness in older subjects (≥12 years of age). In conclusion, our study describes an epidemic of DENV-4 in León, Nicaragua, associated with a novel lineage of genotype GIIb, which contains two amino acid changes not observed in DENV-4 before 2022.

The Centers for Research in Emerging Infectious Diseases (CREID) was established to enhance pandemic preparedness by studying emerging/reemerging pathogens, especially in resource-limited regions. To overcome infrastructure challenges, a low-cost, field-deployable method for extracting total nucleic acids is essential, eliminating reliance on expensive equipment, power, and cold chain systems used in traditional extraction techniques. To address this challenge, we developed an RNA extraction and storage method (RNAES) that meets these criteria. Herein, we report RNAES inactivation efficacy against nine prototype viruses (Middle Eastern respiratory syndrome coronavirus, Japanese encephalitis virus, West Nile virus, Hantaan virus, measles virus, Heartland virus, enterovirus A71, chikungunya virus, and Western equine encephalitis virus) representing seven pandemic potential virus families. We compare the RNAES method to the Qiagen QIAamp kit across various viral loads and field sample types. The presence of infectious virus in RNA samples was quantified using plaque assays. Successful inactivation of viruses was demonstrated for six enveloped virus families spiked into matrices routinely collected at field sites. The seventh family tested (Picornaviridae) was not completely inactivated, likely due to non-enveloped viruses being differentially susceptible to the lysis chemistry of the RNAES kit. The commercial comparator inactivated all viruses tested. Specialized biosafety facilities, specific detailed permits, and comprehensive logistics are required to ensure safety when handling and shipping potentially infectious samples. Inactivating pathogens at the point of collection reduces risks and simplifies sample transfer for critical outbreak research. Confidently ensuring that an isolated nucleic acid sample is non-infectious using RNAES will enable safer, and more efficient downstream analysis.

In 2019–2020, dengue virus (DENV) type 4 emerged to cause the largest DENV outbreak in Paraguay’s history. This study sought to characterize dengue relative to other acute illness cases and use phylogenetic analysis to understand the outbreak’s origin. Individuals with an acute illness (≤7 days) were enrolled and tested for DENV nonstructural protein 1 (NS1) and viral RNA by real-time RT-PCR. Near-complete genome sequences were obtained from 62 DENV-4 positive samples. From January 2019 to March 2020, 799 participants were enrolled: 253 dengue (14 severe dengue, 5.5%) and 546 other acute illness cases. DENV-4 was detected in 238 dengue cases (94.1%). NS1 detection by rapid test was 52.5% sensitive (53/101) and 96.5% specific (387/401) for dengue compared to rRT-PCR. DENV-4 sequences were grouped into two clades within genotype II. No clustering was observed based on dengue severity, location, or date. Sequences obtained here were most closely related to 2018 DENV-4 sequences from Paraguay, followed by a 2013 sequence from southern Brazil. DENV-4 can result in large outbreaks, including severe cases, and is poorly detected with available rapid diagnostics. Outbreak strains seem to have been circulating in Paraguay and Brazil prior to 2018, highlighting the importance of sustained DENV genomic surveillance.

Altered mechanotransduction has been proposed as a putative mechanism for disease pathophysiology, yet evidence remains scarce. Here we introduce a concept we call single cell immuno-mechanical modulation, which links immunology, integrin biology, cellular mechanics, and disease pathophysiology and symptomology. Using a micropatterned hydrogel-laden coverslip compatible with standard fluorescence microscopy, we conduct a clinical mechanobiology study, specifically focusing on immune thrombocytopenia (ITP), an autoantibody-mediated platelet disorder that currently lacks a reliable biomarker for bleeding risk. We discover that in pediatric ITP patients (n = 53), low single platelet contraction force alone is a “physics-based” biomarker of bleeding (92.3% sensitivity, 90% specificity). Mechanistically, autoantibodies and monoclonal antibodies drive increases and decreases of cell force by stabilizing integrins in different conformations depending on the targeted epitope. Hence, immuno-mechanical modulation demonstrates how antibodies may pathologically alter mechanotransduction to cause clinical symptoms and this phenomenon can be leveraged to control cellular mechanics for research, diagnostic, and therapeutic purposes. Altered mechanotransduction has been proposed as a mechanism for disease pathophysiology, yet evidence remains scarce. Here, the authors show that antibodies from patients with bleeding disorders bind to integrins and modulate platelet cell contraction force, and this correlates with clinical symptoms.

Background Dengue is a leading cause of febrile illness among international travellers. We aimed to describe the epidemiology and clinical characteristics of imported dengue in returning travellers evaluated at GeoSentinel sites from 2007 to 2022. Methods We retrieved GeoSentinel records of dengue among travellers residing in non-endemic countries. We considered dengue confirmed when diagnosed by a positive dengue virus (DENV)-specific reverse-transcriptase polymerase chain reaction, positive NS-1 antigen and/or anti-DENV IgG seroconversion, and probable when diagnosed by single anti-DENV IgM or high-titre anti-DENV IgG detection. Severe dengue was defined as evidence of clinically significant plasma leakage or bleeding, organ failure, or shock, according to the 2009 World Health Organization guidance. Complicated dengue was defined as either severe dengue or dengue with presence of any warning sign. Analyses were descriptive. Results This analysis included 5958 travellers with confirmed (n = 4859; 81.6%) or probable (n = 1099; 18.4%) dengue. The median age was 33 years (range: <1-91); 3007 (50.5%) travellers were female. The median travel duration was 21 days (interquartile range [IQR]: 15-32). The median time between illness onset and GeoSentinel site visit was 7 days (IQR: 4-15). The most frequent reasons for travel were tourism (67.3%), visiting friends or relatives (12.2%) and business (11.0%). The most frequent regions of acquisition were South East Asia (50.4%), South Central Asia (14.9%), the Caribbean (10.9%) and South America (9.2%). Ninety-five (1.6%) travellers had complicated dengue, of whom 27 (0.5%) had severe dengue and one died. Of 2710 travellers with data available, 724 (26.7%) were hospitalized. The largest number of cases (n = 835) was reported in 2019. Conclusions A broad range of international travellers should be aware of the risk of acquiring dengue and receive appropriate pre-travel counselling regarding preventive measures. Prospective cohort studies are needed to further elucidate dengue risk by destination and over time, as well as severe outcomes and prolonged morbidity (long dengue) due to travel-related dengue.

Rift Valley fever virus (RVFV) is an adaptable arbovirus that can be transmitted by a wide variety of arthropods. Widespread urban transmission of RVFV has not yet occurred, but peri-urban outbreaks of RVFV have recently been documented in East Africa. We previously reported low-level exposure in urban communities and highlighted the risk of introduction via live animal influx. We deployed a slaughtered animal testing framework in response to an early warning system at two urban slaughterhouses and tested animals entering the meat value chain for anti-RVFV IgG and IgM antibodies. We simultaneously trapped mosquitoes for RVFV and bloodmeal testing. Out of 923 animals tested, an 8.5% IgG seroprevalence was identified but no evidence of recent livestock exposure was detected. Mosquito species abundance varied greatly by slaughterhouse site, which explained 52% of the variance in blood meals. We captured many Culex spp., a known RVFV amplifying vector, at one of the sites (p < 0.001), and this species had the most diverse blood meals. No mosquito pools tested positive for RVFV antigen using a rapid VecTOR test. These results expand understanding of potential RVF urban disease ecology, and highlight that slaughterhouses are key locations for future surveillance, modelling, and monitoring efforts.

We describe the case of a returned traveler to the United States from Ecuador who had an acute febrile illness, initially diagnosed as Oropouche fever. This illness was later confirmed to be a rare infection with Iquitos virus, a related bunyavirus that shares 2 of 3 genome segments with Oropouche virus.

Oropouche virus (OROV) is an arbovirus transmitted by midges that is now emerging outside the Amazon region. For the first time, fatal cases were reported, and possible vertical transmission of OROV is under investigation. Knowledge gaps remain concerning the natural history of OROV infection, host immune response and vector competence.