A rapid, high-throughput, and sensitive PEG-precipitation method for SARS-CoV-2 wastewater surveillance.

The effective application of wastewater surveillance is dependent on testing capacity and sensitivity to obtain high spatial resolution testing results for a timely targeted public health response. To achieve this purpose, the development of rapid, high-throughput, and sensitive virus concentration methods is urgently needed. Various protocols have been developed and implemented in wastewater surveillance networks so far, however, most of them lack the ability to scale up testing capacity or cannot achieve sufficient sensitivity for detecting SARS-CoV-2 RNA at low prevalence. In the present study, using positive raw wastewater in Hong Kong, a PEG precipitation-based three-step centrifugation method was developed, including low-speed centrifugation for large particles removal and the recovery of viral nucleic acid, and medium-speed centrifugation for the concentration of viral nucleic acid. This method could process over 100 samples by two persons per day to reach the process limit of detection (PLoD) of 3286 copies/L wastewater. Additionally, it was found that the testing capacity could be further increased by decreasing incubation and centrifugation time without significantly influencing the method sensitivity. The entire procedure uses ubiquitous reagents and instruments found in most laboratories to obtain robust testing results. This high-throughput, cost-effective, and sensitive tool will promote the establishment of nearly real-time wastewater surveillance networks for valuable public health information.

Characteristics and performances of microalgal-bacterial consortia in a mixture of raw piggery digestate and anoxic aerated effluent.

The piggery digestate of high ammonia was mixed with the anoxic aerated effluent of high nitrate and phosphorus, to cultivate a microalgal-bacterial consortium for simultaneous pollution removal and resource recovery. The highest removal of total inorganic nitrogen was achieved at 324.77 mg/L in 40% piggery digestate mixed with 60% anoxic aerated effluent, along with the most microalgae biomass production. The crude protein and fatty acids of C14-C20 in microalgae cells were 21.80% and 69.78%, indicating that this mixing strategy could produce abundant microalgal biomass suitable for biofuel generation and animal feed. High-throughput sequencing showed that microbial diversity increased and Paenibacillus, Thiopseudomonas and Pseudomonas were the dominant species promoting microalgal growth. Overall, these results provided a new insight of mixing two types of wastewaters for cultivating microalgal-bacterial consortia, to remove contamination and recover nutrients simultaneously.