Blood from all four birds tested positive by PCR for BFDV, and analysis of the recovered full BFDV genomes identified them as belonging to the BFDV-A strain.

The antimicrobial additive triclosan has been used in personal care products widely across the globe for decades. Triclosan resistance has been noted among Vibrio spp., but reports have been anecdotal and the extent of phenotypic triclosan resistance across the Vibrionaceae family has not been established. Here, triclosan resistance was determined for Vibrionaceae strains across nine distinct clades. Minimum inhibitory concentrations (MIC) were determined for 70 isolates from clinical (n = 6) and environmental sources (n = 64); only two were susceptible to triclosan. The mean MIC for all resistant Vibrionaceae was 53 µg mL−1 (range 3.1–550 µg mL−1), but was significantly different between clades (p < 0.001). The highest mean triclosan MIC was observed in the Splendidus clade (200 µg mL−1; n = 3). Triclosan mean MICs were 68.8 µg mL−1 in the Damselae clade and 45.3 µg mL−1 in the Harveyi clade. The lowest mean MIC was observed in the Cholerae clade with 14.4 µg mL−1, which was primarily represented by clinical strains. There were no significant differences in triclosan MIC among individual species or among environmental strains isolated from different locations. Overall, phenotypic triclosan resistance appears to be widespread across multiple clades of Vibrionaceae.

Wastewater surveillance has proven to be an effective tool to monitor the transmission and emergence of infectious agents at a community scale. Workflows for wastewater surveillance generally rely on concentration steps to increase the probability of detection of low-abundance targets, but pre-concentration can substantially increase the time and cost of analyses while also introducing additional loss of target during processing. To address some of these issues, we conducted a longitudinal study implementing a simplified workflow for SARS-CoV-2 detection from wastewater, using a direct column-based extraction approach. Composite influent wastewater samples were collected weekly for one year between June 2020 and June 2021 in Athens-Clarke County, Georgia, USA. Bypassing any concentration step, low volumes (280 µL) of influent wastewater were extracted using a commercial kit, and immediately analyzed by RT-qPCR for the SARS-CoV-2 N1 and N2 gene targets. SARS-CoV-2 viral RNA was detected in 76% (193/254) of influent samples, and the recovery of the surrogate bovine coronavirus was 42% (IQR: 28%, 59%). N1 and N2 assay positivity, viral concentration, and flow-adjusted daily viral load correlated significantly with per-capita case reports of COVID-19 at the county-level (ρ = 0.69 to 0.82). To compensate for the method's high limit of detection (approximately 106–107 copies L−1 in wastewater), we extracted multiple small-volume replicates of each wastewater sample. With this approach, we detected as few as five cases of COVID-19 per 100 000 individuals. These results indicate that a direct-extraction-based workflow for SARS-CoV-2 wastewater surveillance can provide informative and actionable results.