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National Committee on Marine Sciences (NCMS)

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    Multifaceted assessment of wastewater-based epidemiology for SARS-CoV-2 in selected urban communities in Davao City, Philippines: A pilot study
    Otero, Maria Catherine B.; Murao, Lyre Anni E.; Limen, Mary Antoinette G.; Caalim, Daniel Rev A.; Gaite, Paul Lorenzo A.; Bacus, Michael G.; Acaso, Joan T.; Miguel, Refeim M.; Corazo, Kahlil; Knot, Ineke E.; Sajonia, Homer; de los Reyes, Francis L.; Jaraula, Caroline Marie B.; Baja, Emmanuel S.; Del Mundo, Dann Marie N. (MDPI, 2022-07-19)
    Over 60 countries have integrated wastewater-based epidemiology (WBE) in their COVID-19 surveillance programs, focusing on wastewater treatment plants (WWTP). In this paper, we piloted the assessment of SARS-CoV-2 WBE as a complementary public health surveillance method in susceptible communities in a highly urbanized city without WWTP in the Philippines by exploring the extraction and detection methods, evaluating the contribution of physico-chemical–anthropogenic factors, and attempting whole-genome sequencing (WGS). Weekly wastewater samples were collected from sewer pipes or creeks in six communities with moderate-to-high risk of COVID-19 transmission, as categorized by the City Government of Davao from November to December 2020. Physico-chemical properties of the wastewater and anthropogenic conditions of the sites were noted. Samples were concentrated using a PEG-NaCl precipitation method and analyzed by RT-PCR to detect the SARS-CoV-2 N, RdRP, and E genes. A subset of nine samples were subjected to WGS using the Minion sequencing platform. SARS-CoV-2 RNA was detected in twenty-two samples (91.7%) regardless of the presence of new cases. Cycle threshold values correlated with RNA concentration and attack rate. The lack of a sewershed map in the sampled areas highlights the need to integrate this in the WBE planning. A combined analysis of wastewater physico-chemical parameters such as flow rate, surface water temperature, salinity, dissolved oxygen, and total dissolved solids provided insights on the ideal sampling location, time, and method for WBE, and their impact on RNA recovery. The contribution of fecal matter in the wastewater may also be assessed through the coliform count and in the context of anthropogenic conditions in the area. Finally, our attempt on WGS detected single-nucleotide polymorphisms (SNPs) in wastewater which included clinically reported and newly identified mutations in the Philippines. This exploratory report provides a contextualized framework for applying WBE surveillance in low-sanitation areas.
    The authors thank the Davao City Health Office, the local government units under the City Government of Davao, the partner hospitals for their support and assistance, and Diana Aga for discussions on sample collection and analyses. The authors would also like to thank the members of the Bortz Virology Laboratory at the University of Alaska Anchorage- Ralf Dagdag and Matthew Redlinger, Amanda Warr from the Roslin Institute, Nicole Wheeler from the University of Birmingham, Lara Urban, co-founder of PuntSeq, and Joe Russell from MRI Global for their expert advice in nanopore and wastewater sequencing.
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    BFAR Administrative Circular No. 256: Series of 2015. Declaring malachite green and gentian violet as health hazards and prohibiting their use in food fish production and trade.
    (Bureau of Fisheries and Aquatic Resources, 2015-02-12)
    This Circular, consisting of 25 Sections divided into seven Chapters and one Annex, prohibits the use of malachite green and gentian violet in all stages of aquaculture production of fishery and aquatic products intended for human consumption and the distribution for commerce of fishery and aquatic products derived from aquaculture containing traces of the dyes. It shall be unlawful for any person to: 1) Use malachite green or gentian violet in the aquaculture production of food fish; 2) Produce, handle or process for sale, offer for sale, distribute in commerce fish or fishery product bearing or containing malachite green or gentian violet; and, 3) Import into the Philippines or export fish or fishery product bearing or containing malachite green or gentian violet. Any importer, exporter, aquaculture/farm operator, or distributor found violating any of the provisions of this Circular shall be subject to the following administrative sanctions: a) for the first offense, suspension for three months of the registration as importer, exporter, aquaculture/farm operator or distributor; b) for the second offense, six-month suspension of the registration; and, c) for the third offense, revocation of the registration and delisting.