Wastewater-based epidemiology (WBE) is a valuable tool for monitoring pathogen spread in communities; however, current protocols mainly target non-enveloped viruses. This study addresses the need for standardized methods to detect both enveloped and non-enveloped viruses by testing four aluminum hydroxide adsorption–precipitation techniques. Wastewater samples were spiked with an enveloped virus surrogate (Φ6 bacteriophage) and a non-enveloped virus surrogate (MS2 coliphage), and viral recovery was assessed using reverse-transcription quantitative PCR (RT-qPCR). The highest recovery for the enveloped virus was achieved with AlCl₃ at pH 3.5, a 15 min flocculation time, and a 3% elution solution concentration. For the non-enveloped virus, optimal recovery was found with AlCl₃ at pH 6.0, no flocculation time, and a 10% elution solution. The best method for recovering both virus types used AlCl₃ at pH 6.0, 15 min flocculation, and a 3% elution solution concentration. This study shows that while optimal conditions vary between virus types, a standardized AlCl₃ flocculation protocol can efficiently recover both, providing a cost-effective approach for outbreak monitoring in Grenada.

Antimicrobial resistance (AMR) is a growing global health threat. This study investigated antibiotic resistance in E. coli isolates from municipal wastewater (86 isolates) and clinical urinary tract infection (UTI) cases (34 isolates) in a Grenadian community, using data from January 2022 to October 2023. Antibiogram data, assessed per WHO guidelines for Critically Important antimicrobials (CIA), showed the highest resistance levels in both clinical and wastewater samples for ampicillin, followed by amoxicillin/clavulanic acid and nalidixic acid, all classified as Critically Important. Similar resistance was observed for sulfamethoxazole-trimethoprim (highly important) in both groups, with nitrofurantoin showing resistance in the important category. According to the WHO AWaRe classification, ampicillin (ACCESS group) had the highest resistance, while nitrofurantoin had the lowest across all samples. The WATCH group antibiotics, cefuroxime and cefoxitin, showed comparable resistance levels, whereas aztreonam from the RESERVE group (tested only in wastewater) was 100% sensitive. Multiple Antibiotic Resistance (MAR) index analysis revealed that 7% of wastewater and 38.2% of clinical samples had MAR values over 0.2, indicating prior antibiotic exposure in clinical isolates. These parallel patterns in wastewater and clinical samples highlight wastewater monitoring as a valuable tool for AMR surveillance, supporting antibiotic stewardship through ongoing environmental and clinical assessment.

Wastewater-based epidemiology (WBE) allows for early surveillance of viral pathogens, including SARS-CoV-2. Simplified low-cost approaches are needed to deploy WBE surveillance in resource-limited small-island settings, where high sensitivity must be maintained. In this study, we optimized key upstream steps in an electronegative membrane virus adsorption–elution (VIRADEL) workflow, including sample acidification, composite sampling duration, and RT-qPCR inhibition mitigation. Wastewater influent was sampled at a pump station in Grenada using 12 h and 24 h time-weighted composite samples, concentrated using electronegative membrane VIRADEL with and without sample acidification (pH 3.5), and used Phi 6 (enveloped virus) and MS2 (non-enveloped virus) bacteriophages as process controls and PMMoV as a fecal-derived normalization target. Targets for SARS-CoV-2 N1 and a non-enveloped virus surrogate were measured by RT-qPCR. Quantitative wastewater data were compared to reported clinical cases in the community. Sample acidification significantly increased recovery of the enveloped process control, Phi 6 (p < 0.01) indicating improved efficiency in capturing enveloped viral targets during filtration. Twelve-hour composite samples had a false-negative percentage of 88%, while 24 h samples had only 6% false negatives and were able to mirror clinical case trends. Wastewater viral signals were detected 3–5 days prior to an increase in clinical cases. Hydraulic travel time within the contributing sewer network was not directly measured; therefore, the reported 3–5 day lead time reflects the combined effect of shedding dynamics, sampling integration, and sewer transport. This optimized workflow was deployed for nine months showing sustained analytical performance and operational feasibility.