National Committee on Marine Sciences (NCMS)
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- Fisheries Administrative Order No. 234: Series of 2010. Accreditation of marine biotoxin testing centers.(Department of Agriculture, 2010-04-28)
- Characterization of Alexandrium tamutum (Dinophyceae) isolated from Philippine waters, with the rare detection of paralytic shellfish toxinBenico, Garry; Azanza, Rhodora (Association of Systematic Biologists of the Philippines, 2022-04-01)Alexandrium tamutum M.Montressor, A.Beran & U.John is a non-toxic, bloom-forming dinoflagellate species commonly reported in temperate waters. In this study, 8 cultures of A. tamutum established from Bolinao Channel and Manila Bay, Philippines were characterized in terms of their morphology, phylogeny and toxicity. Cells were roundish, measuring 25.5 –29.84 µm long and 26.2–28.45 µm wide. The nucleus is equatorially elongated and located at the center of the cell. The chloroplasts are numerous, golden brown in color and radially arranged. Thecal tabulation is typical of Alexandrium: APC, 4', 6'', 6c, 6s, 5''', 2''''. Shape of the taxonomically informative thecal plates such as sixth precingular plate (6'') and posterior sulcal plate (sp) was similar to A. tamutum, which confirms the species identity. However, the presence of anterior and posterior attachment pores observed in our cultured isolates is the first case in this species. Molecular phylogeny inferred from LSU rDNA and ITS supports our identification by forming a well-supported clade composed of A. tamutum strains from other geographic regions. HPLC analysis showed that A. tamutum is generally non-toxic except for strain ATC9 which has low amount of decarbamoylsaxitoxin (dcSTX), resulting to a toxicity of 0.07 fmole STX eq per cell. The present study reports the first verified occurrence of Philippine A. tamutum with reliable morphological and molecular information, including the first record in Manila Bay and first detection of PST in one strain at a certain culture period.We acknowledge the Department of Science and Technology Philippine Council for Agriculture, Aquatic and Natural Resources Research and Development (DOST PCAARRD) and the University of the Philippines-The Marine Science Institute (UP-MSI) for the funding support. We are grateful for the assistance of Joshua Vacarizas, Keith Pinto and Jenelyn Mendoza for the molecular and toxicity analyses of the cultures. We also acknowledge Estrelita Flores, Emelita Eugenio and Jayson Orpeza for their assistance during the fieldwork and other logistical support.
- Connections and clustering of Paralytic Shellfish Toxin events among coastal embayments in an archipelago partly mediated by advectionPunongbayan, Andalus T.; Wang, Ysabel D.; Villanoy, Cesar L.; Yñiguez, Aletta T. (Elsevier, 2022-01)The potential for advection to influence harmful algal bloom (HAB) spread in adjacent embayments and islands has not been investigated in the Philippines as previous studies have focused on HAB dynamics within specific embayments. Advection of HABs may be relevant in the Samar-Leyte region where adjacent sites are subject to recurrent blooms of Pyrodinium bahamense. We used different analyses to extract the potential role of advection in blooms in the region. First, we used Bayesian and information theoretic approaches applied to historical data on shellfish bans to quantify spatial dependencies in HAB occurrences between sites. Then, to determine whether such dependencies are related to circulation patterns in the region, we analyzed connectivity using a hydrodynamic model coupled with a conservative tracer-based HAB model. The Bayesian network showed that in 7 out of 11 sites, the probability of a shellfish ban depended on the state of an adjacent site. Site pairs with direct dependence relations also shared relatively high similarity in HAB occurrences over time. In the modelled network, bans tend to occur sequentially, spreading from a few sites with relatively high probabilities for ban events. A subset of sites (sources) were found to be informative of future HAB event probabilities in other sites (destinations) over time lags that are generally longer the farther the destination. Modelled surface advection showed high connectivity strengths between sources and destinations associated with circulation features, e.g., an anticyclonic current in Leyte, wind-driven coastal current in western Samar, and tidally-driven flow in the shallow embayments in southwest Samar. High connectivities were correlated with direct dependence relations in the Bayesian network. Connectivity explained up to about 1/3 of the variance in statistical dependencies between ban signals. Our results show that Paralytic Shellfish Toxin events within this region can be due to advection of blooms originating from nearby areas rather than localized cyst dynamics within an embayment. Particular sites were also identified as potential indicator sites within spatial clusters for monitoring since they are typically where blooms originate and spread from.
- Global mass spectrometric analysis reveals chemical diversity of secondary metabolites and 44-Methylgambierone production in Philippine Gambierdiscus strainsMalto, Zabrina Bernice L.; Benico, Garry A.; Batucan, Jeremiah D.; Dela Cruz, James; Romero, Marc Lawrence J.; Azanza, Rhodora V.; Salvador-Reyes, Lilibeth A. (Frontiers Media SA, 2022-02-04)Surveillance and characterization of emerging marine toxins and toxigenic dinoflagellates are warranted to evaluate their associated health risks. Here, we report the occurrence of the ciguatera poisoning-causative dinoflagellate Gambierdiscus balechii in the Philippines. Toxin production and chemical diversity of secondary metabolites in G. balechii GtoxSAM092414, G. balechii Gtox112513, and the recently reported Gambierdiscus carpenteri Gam1BOL080513 were assessed using targeted and untargeted UPLC-MS/MS analysis and radioligand receptor-binding assay (RBA). 44-methylgambierone was produced by all three strains, albeitwith different levels based on RBA and UPLC-HRMS/MS analysis. The fatty acid composition was similar in all strains, while subtle differences in monosaccharide content were observed, related to the collection site rather than the species. Molecular networking using the GNPS database identified 45 clusters belonging to at least ten compound classes, with terpene glycosides, carbohydrate conjugates, polyketides, and macrolides as major convergence points. Species-specific peptides and polyhydroxylated compounds were identified in G. balechii GtoxSAM092414 and G. carpenteri Gam1BOL080513, respectively. These provide a glimpse of the uncharacterized biosynthetic potential of benthic dinoflagellates and highlight the intricate and prolific machinery for secondary metabolites production in these organisms.We would like to thank H. Junio and the Secondary Metabolites Profiling Laboratory of the Institute of Chemistry, University of the Philippines Diliman and K. B. Davis for assistance in the conduct of this study.
- Using constellation pharmacology to characterize a novel α-conotoxin from Conus ateralbusNeves, Jorge L. B.; Urcino, Cristoval; Chase, Kevin; Dowell, Cheryl; Hone, Arik J.; Morgenstern, David; Chua, Victor M.; Ramiro, Iris Bea L.; Imperial, Julita S.; Leavitt, Lee S.; Phan, Jasmine; Fisher, Fernando A.; Watkins, Maren; Raghuraman, Shrinivasan; Tun, Jortan O.; Ueberheide, Beatrix M.; McIntosh, J. Michael; Vasconcelos, Vitor; Olivera, Baldomero M.; Gajewiak, Joanna (MDPI, 2024-02-29)The venom of cone snails has been proven to be a rich source of bioactive peptides that target a variety of ion channels and receptors. α-Conotoxins (αCtx) interact with nicotinic acetylcholine receptors (nAChRs) and are powerful tools for investigating the structure and function of the various nAChR subtypes. By studying how conotoxins interact with nAChRs, we can improve our understanding of these receptors, leading to new insights into neurological diseases associated with nAChRs. Here, we describe the discovery and characterization of a novel conotoxin from Conus ateralbus, αCtx-AtIA, which has an amino acid sequence homologous to the well-described αCtx-PeIA, but with a different selectivity profile towards nAChRs. We tested the synthetic αCtx-AtIA using the calcium imaging-based Constellation Pharmacology assay on mouse DRG neurons and found that αCtx-AtIA significantly inhibited ACh-induced calcium influx in the presence of an α7 positive allosteric modulator, PNU-120596 (PNU). However, αCtx-AtIA did not display any activity in the absence of PNU. These findings were further validated using two-electrode voltage clamp electrophysiology performed on oocytes overexpressing mouse α3β4, α6/α3β4 and α7 nAChRs subtypes. We observed that αCtx-AtIA displayed no or low potency in blocking α3β4 and α6/α3β4 receptors, respectively, but improved potency and selectivity to block α7 nAChRs when compared with αCtx-PeIA. Through the synthesis of two additional analogs of αCtx-AtIA and subsequent characterization using Constellation Pharmacology, we were able to identify residue Trp18 as a major contributor to the activity of the peptide.We thank the Universidade Técnica do Atlântico (UTA; Cabo Verde) and Cabo Verde National Directorate of the Environment for sample collection support. We thank David Ginty (Harvard Univ.) for sharing the transgenic mice. We thank Joseph W. Aman for conducting the initial screening of the venom fractions, Samuel Espino for providing us with the picture of the shells, and Grzegorz Gajewiak for assistance with handling the photos and preparing some of the graphics used in this article.