A molecular surveillance-guided vector control response to concurrent dengue and West Nile virus outbreaks in a COVID-19 hotspot of Florida (preprint)

Simultaneous dengue virus (DENV) and West Nile virus (WNV) outbreaks in Florida, USA, in 2020 resulted in 71 dengue virus serotype 1 and 86 WNV human cases. Our outbreak response leveraged a molecular diagnostic screen of mosquito populations for DENV and WNV in Miami-Dade County to quickly employ targeted mosquito abatement efforts. We detected DENV serotypes 2 and 4 in mosquito pools, highlighting the silent circulation of diverse dengue serotypes in mosquitoes. Additionally, we found WNV-positive mosquito pools in areas with no historical reports of WNV transmission. These findings demonstrate the importance of proactive, strategic arbovirus surveillance in mosquito populations to prevent and control outbreaks, particularly when other illnesses (e.g., COVID-19), which present with similar symptoms are circulating concurrently. Growing evidence for substantial infection prevalence of dengue in competent mosquito vectors in the absence of local index cases suggests a higher level of dengue endemicity in Florida than previously thought. Article Summary LineEvidence of increasing dengue endemicity in Florida: Vector surveillance during dengue and West Nile virus outbreaks revealed widespread presence of other dengue virus serotypes in the absence of local index cases.

Niclosamide reverses SARS-CoV-2 control of lipophagy (preprint)

The global effort to combat COVID-19 rapidly produced a shortlist of approved drugs with antiviral activities for clinical repurposing. However, the jump to clinical testing was lethal in some cases as a full understanding of the mechanism of antiviral activity as opposed to pleiotropic activity/toxicity for these drugs was lacking. Through parallel lipidomic and transcriptomic analyses we observed massive reorganization of lipid profiles of infected Vero E6 cells, especially plasmalogens that correlated with increased levels of virus replication. Niclosamide (NIC), a poorly soluble anti-helminth drug identified for repurposed treatment of COVID-19, reduced the total lipid profile that would otherwise amplify during virus infection. NIC treatment reduced the abundance of plasmalogens, diacylglycerides, and ceramides, which are required for virus pro-duction. Future screens of approved drugs may identify more druggable compounds than NIC that can safely but effectively counter SARS-CoV-2 subversion of lipid metabolism thereby reducing virus replication. However, these data support the consideration of niclosamide as a potential COVID-19 therapeutic given its modulation of lipophagy leading to the reduction of virus egress and the subsequent regulation of key lipid mediators of pathological inflammation.

Earliest Detection to Date of SARS-CoV-2 in Florida: Identification Together With Influenza Virus on the Main Entry Door of a University Building, February 2020 (preprint)

BackgroundQuestions persist about patterns of initial dissemination of SARS-CoV-2 in the United States in early 2020.MethodsIn February and March, 2020, environmental surface swab samples were collected from the handle of the main entry door of a major university building in Florida, as part of a pilot surveillance project screening for influenza. Samples were taken at the end of regular classroom hours, between the dates of February 1-5 and February 19-March 4, 2020. ResultsInfluenza H1N1pdm09 was isolated from the door handle on four of the 19 days sampled.  Both SARS-CoV-2 and influenza virus were detected in the sample collected on February 21, 2020. Based on sequence analysis, the Florida SARS-CoV-2 strain (designated UF-11) was identical to strains being identified in Washington state during the same time period, while the earliest similar sequences were sampled in China/Hubei between Dec 30th 2019 and Jan 5th 2020. The first human case of COVID-19 was not officially reported in Florida until March 1st. In an analysis of sequences from COVID-19 patients in this region of Florida, there was only limited evidence of subsequent dissemination of the UF-11 strain.  Identical or highly similar strains, possibly related through a common transmission chain, were detected with increasing frequency in Washington state between end of February and beginning of March. ConclusionsOur data provide further documentation of the rapid early spread of SARS-CoV-2, and underscore the likelihood that closely related strains were cryptically circulating in multiple U.S. communities before the first “official” cases were recognized.

Viable SARS-CoV-2 in the air of a hospital room with COVID-19 patients (preprint)

BackgroundThere currently is substantial controversy about the role played by SARS-CoV-2 in aerosols in disease transmission, due in part to detections of viral RNA but failures to isolate viable virus from clinically generated aerosols. MethodsAir samples were collected in the room of two COVID-19 patients, one of whom had an active respiratory infection with a nasopharyngeal (NP) swab positive for SARS-CoV-2 by RT-qPCR. By using VIVAS air samplers that operate on a gentle water-vapor condensation principle, material was collected from room air and subjected to RT-qPCR and virus culture. The genomes of the SARS-CoV-2 collected from the air and of virus isolated in cell culture from air sampling and from a NP swab from a newly admitted patient in the room were sequenced. FindingsViable virus was isolated from air samples collected 2 to 4.8m away from the patients. The genome sequence of the SARS-CoV-2 strain isolated from the material collected by the air samplers was identical to that isolated from the NP swab from the patient with an active infection. Estimates of viable viral concentrations ranged from 6 to 74 TCID50 units/L of air. InterpretationPatients with respiratory manifestations of COVID-19 produce aerosols in the absence of aerosol-generating procedures that contain viable SARS-CoV-2, and these aerosols may serve as a source of transmission of the virus. FundingPartly funded by Grant No. 2030844 from the National Science Foundation and by award 1R43ES030649 from the National Institute of Environmental Health Sciences of the National Institutes of Health, and by funds made available by the University of Florida Emerging Pathogens Institute and the Office of the Dean, University of Florida College of Medicine. Research in contextO_ST_ABSEvidence before this studyC_ST_ABSVarious studies report detection of SARS-CoV-2 in material collected by air samplers positioned in clinics and in some public spaces. For those studies, detection of SARS-CoV-2 has been by indirect means; instead of virus isolation, the presence of the virus in material collected by air samplers has been through RT-PCR detection of SARS-CoV-2 RNA. However, questions have been raised about the clinical significance of detection of SARS-CoV-2 RNA, particularly as airborne viruses are often inactivated by exposure to UV light, drying, and other environmental conditions, and inactivated SARS-CoV-2 cannot cause COVID-19. Added value of this studyOur virus isolation work provides direct evidence that SARS-CoV-2 in aerosols can be viable and thus pose a risk for transmission of the virus. Furthermore, we show a clear progression of virus-induced cytopathic effects in cell culture, and demonstrate that the recovered virus can be serially propagated. Moreover, we demonstrate an essential link: the viruses we isolated in material collected in four air sampling runs and the virus in a newly admitted symptomatic patient in the room were identical. These findings strengthen the notion that airborne transmission of viable SARS-CoV-2 is likely and plays a critical role in the spread of COVID-19. Implications of all the available evidenceScientific information on the mode of transmission should guide best practices Current best practices for limiting the spread of COVID-19. Transmission secondary to aerosols, without the need for an aerosol-generating procedure, especially in closed spaces and gatherings, has been epidemiologically linked to exposures and outbreaks. For aerosol-based transmission, measures such as physical distancing by 6 feet would not be helpful in an indoor setting and would provide a false-sense of security. With the current surges of cases, to help stem the COVID-19 pandemic, clear guidance on control measures against SARS-CoV-2 aerosols are needed.