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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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    Feeding and reproductive phenotypic traits of the sea urchin Tripneustes gratilla in seagrass beds impacted by eutrophication
    Bangi, Helen Grace P.; Juinio-Meñez, Marie Antonette (MDPI AG, 2023-07-11)
    The sea urchin Tripneustes gratilla is a major grazer and is, hence, an excellent key model organism to study to gain a better understanding of responses to changes in its habitat. We investigated whether there are significant variations in the feeding and reproductive phenotypic traits of populations from three seagrass bed sites, with respect to their proximity to fish farms in Bolinao, northwestern Philippines. We established three stations in each of the three sites: the far, the intermediate, and those near the fish farms, and compared the sea urchins’ phenotypic traits and determined whether these were related to seagrass productivity and water parameters. Regardless of the sampling period, adult sea urchins (66.92 ± 0.27 mm test diameter, TD, n = 157) from the areas intermediate and near to the fish farms had significantly lower indices of Aristotle’s lantern, gut contents, gut and gonads, and lower gonad quality (high percentage of unusual black gonads), compared to those from the far stations. Multivariate analysis showed that the smaller feeding structures and gut, lower consumption rates and lower gonad indices and quality of sea urchins in the intermediate and near fish farms were positively related to lower shoot density, leaf production and species diversity, as well as lower water movement in those stations. The larger size of the Aristotle’s lantern in the far stations was not related to food limitations. More importantly, the phenotypic variability in the feeding structures and gonads of sea urchins in the same seagrass bed provides new evidence regarding the sensitivity of this species to environmental factors that may affect variability in food quality.
    The authors are very grateful to the anonymous reviewers for providing significant comments and suggestions to improve the manuscript. They are also indebted to the following: Rene R. Rollon, for providing advice on seagrass sampling techniques; Symon Dworjanyn, for providing valuable inputs in the earlier version of this manuscript; Marilou San Diego-McGlone, for providing some water quality data in Bolinao; Charissa M. Ferrerra, for the assistance provided on the Ocean Data View mapping software; Ma Josefa R. Pante, for some statistical advice. The authors would like to thank Jay R Gorospe for reviewing and providing valuable suggestions on the revised version of the manuscript, likewise to Lambert Meñez, for critically editing the manuscript, and to Jerwin Baure for additional assistance in copy editing the manuscript. The authors are thankful to Larry Milan, Jack Rengel, Lawrence Ramoran, for assisting the authors in field sampling and laboratory processing of samples. L. Milan, Jan Noelle Rimando and Aphrodite Entoma assisted in laboratory analysis of samples, particularly in gut content analysis.