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Bunyaviruses are significant human pathogens, causing diseases ranging from hemorrhagic fevers to encephalitis. Among these viruses, La Crosse virus (LACV), a member of the California serogroup, circulates in the eastern and midwestern United States. While LACV infection is often asymptomatic, dozens of cases of encephalitis are reported yearly. Unfortunately, no antivirals have been approved to treat LACV infection. Here, we developed a method to rapidly test potential antivirals against LACV infection. From this screen, we identified several potential antiviral molecules, including known antivirals. Additionally, we identified many novel antivirals that exhibited antiviral activity without affecting cellular viability. Valinomycin, a potassium ionophore, was among our top targets. We found that valinomycin exhibited potent anti-LACV activity in multiple cell types in a dose-dependent manner. Valinomycin did not affect particle stability or infectivity, suggesting that it may preclude virus replication by altering cellular potassium ions, a known determinant of LACV entry. We extended these results to other ionophores and found that the antiviral activity of valinomycin extended to other viral families, including bunyaviruses (Rift Valley fever virus, Keystone virus), enteroviruses (coxsackievirus, rhinovirus), flavirivuses (Zika virus), and coronaviruses (human coronavirus 229E [HCoV-229E] and Middle East respiratory syndrome CoV [MERS-CoV]). In all viral infections, we observed significant reductions in virus titer in valinomycin-treated cells. In sum, we demonstrate the importance of potassium ions to virus infection, suggesting a potential therapeutic target to disrupt virus replication., (Copyright © 2020 American Society for Microbiology.)

During emergencies in the United States, the Environmental Protection Agency (EPA) currently recommends using bottled water, or boiling or treating water by adding 1/8 teaspoon (or 8 drops) of bleach to 1 gal of water. This bleach recommendation is internally inconsistent, a relatively high chlorine dose (5.55-8.67 mg/L), and unsupported by evidence. In this study, bleach was added in three different dosages to six waters available to emergency-affected populations in each of six states; free chlorine residual (FCR) and Escherichia coli/total coliforms were measured 1-24 h after treatment. Data were analyzed using four efficacy criteria. Results indicated the dosages in the current EPA recommendation are unnecessarily high to ensure (1) maintenance of FCR for 24 h after treatment, (2) absence of E. coli/total coliforms, and (3) establishment of a CT-factor sufficient to inactivate Giardia lamblia and enteric viruses 1 h after treatment. Additionally, emergency-prone populations did not have the materials to complete treatment with bleach in their household. Therefore, we recommend EPA review and revise the current recommendation to establish an internally consistent, criteria-based recommendation that is usable by emergency-affected populations. We also recommend investigating the use of new or commercially available water treatment products for emergency response in the United States.

Information on the required chlorine dose or Ct value (concentration of free chlorine multiplied by contact time) is limited to only a few enteroviruses. In this study the Ct values of some of the reported more chlorine resistant enteroviruses were determined. The Efficiency Hom Model was used to predict the times for 2, 3. and 4 -log inactivation of echovirus 1 and 12, coxsackievirus B5 and poliovirus type 1 at pH 7.5 and 9 at 5 degrees C. Coxsackievirus B5 was the most resistant to chlorine with a Ct requirement of 11.5 mg x min/L at pH 7.5 at 5 degrees C compared to polio with a Ct of 5.3 mg x min/L under the same conditions. All the viruses were more resistant than polio 1 at pH 9.0.

Enhanced coagulation was evaluated for removal efficacy of coxsackievirus and echovirus (Contaminant Candidate List [CCL] enteroviruses), poliovirus, four potential surrogate bacteriophages, and dissolved organic carbon (DOC). Viruses and DOC were effectively removed using enhanced coagulation, with removals generally improving as dose increased and pH decreased. Optimal enhanced coagulation conditions of 40 mg/L FeCl3 and pH between 5 and 6.5 resulted in a maximum removal of 3.0 logs of coxsackievirus B6, 1.75 logs of echovirus 12, 2.5 logs of poliovirus 1, 1.8 logs of fr, 1.3 logs of phi-X174, 0.36 logs of MS2, 0.29 logs of PRD1, and 41% DOC. Bacteriophages fr and phi-X174 appear to be the most representative surrogates for the physical removal of coxsackievirus, while MS2 and PRD1 are more conservative. For echovirus, MS2 and PRD1 appearto bethe most appropriate surrogates. The relative removal profiles of the enteroviruses (greatest removal of coxsackievirus followed by poliovirus and then echovirus) suggest that studies of the physical removal of poliovirus may be extended to the CCL enteroviruses. These results contribute to evaluations of the CCL and regulatory status of coxsackievirus and echovirus and aid in building a database of the treatment efficiencies of enteroviruses and their surrogates.