Journal Articles - UP - MSI
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- In Silico supported nontarget analysis of contaminants of emerging concern: Increasing confidence in unknown identification in wastewater and surface watersAngeles, Luisa F.; Halwatura, Lahiruni M.; Antle, Jonathan P.; Simpson, Scott; Jaraula, Caroline M.B.; Aga, Diana S. (American Chemical Society, 2021-08-01)Nontarget analysis using liquid chromatography–high resolution mass spectrometry (LC–HRMS) is a valuable approach in characterizing for contaminants of emerging concern (CECs) in the environment. In this study, wastewater and surface water samples from three locations in Manila, Philippines were analyzed for CECs using a nontarget analysis approach with an LC-Orbitrap. A previously optimized semiautomated workflow was used for data processing with Compound Discoverer. A total of 157 compounds were identified, with 21 confirmed with reference standards, 83 confirmed with evidence from a mass spectral library (mzCloud), and 53 tentatively identified using in silico fragmentation (MetFrag). These compounds include pharmaceuticals such as antibiotics, antifungal, and antihypertensive compounds, human metabolites, natural products, pesticides, and industrial chemicals. Compounds confirmed with reference standards include antibiotics ciprofloxacin, clarithromycin, acetyl-sulfamethoxazole, and trimethoprim (2 to 19 ng/L), and antifungal compounds carbendazim and climbazole (3–47 ng/L). The pesticides diethyltoluamide (DEET) and diuron were also detected (37 ng/L). The utility of a preliminary multivariable linear regression quality structure-retention relationship (QSRR) model based on quantum chemical molecular descriptors is demonstrated. This study demonstrates the importance of using tools and software that are helpful for annotating HRMS data and reporting detections according to a standardized classification system. The detection of several CECs in wastewater and surface water samples show the importance of performing nontarget analysis in determining occurrence of CECs in the environment.We acknowledge support from the National Science Foundation PIRE-HEARD award number 1545756 and USAID PEER subaward number 2000009924. S.S. thanks the National Science Foundation (Award #1904825) for support of this research, and the Donors of the American Chemical Society Petroleum Research Fund (PRF-58954-UNI5). We also thank Shyrill Mae Mariano from the Marine Science Institute in the University of the Philippines − Diliman who helped with the sample collection and Rebecca Dickman from University at Buffalo for her help on technical work.
- 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.
- Community structure of periphytic diatoms in early-stage marine biofilms in a mariculture-impacted areaBaure, Jerwin; San Diego-McGlone, Maria Lourdes; Juinio–Meñez, Marie Antonette (Science and Technology Information Institute, 2022-07-06)Periphytic diatoms constitute a major part of complex unicellular aggregations called marine biofilm or periphyton, of which little is known about in tropical marine environments. The early-stage community structure of periphytic diatom genera on two artificial substrates – glass slide and polycarbonate sheets was characterized. These artificial substrates were placed underwater for 6 d and examined on Days 1, 3, and 6 in four sites around Santiago Island, Bolinao, Pangasinan, Philippines, with different relative distances from the intensive milkfish mariculture area. Correlations of diatom community structures in these sites with physico-chemical factors were investigated. The five most abundant diatom genera – namely, Cylindrotheca, Nitzschia, Navicula, Amphora, and Pleurosigma – exhibited a weak correlation with nutrients. Less abundant genera Pseudonitzschia, Haslea, Bacillaria, Thalassionema, Rhizosolenia, Eucampia, Diploneis, Asterionellopsis, Chaetoceros, Bacteriastrum, Licmophora, Skeletonema, Lioloma, Thalassiosira, and Thalassiothrix showed a positive correlation with nutrients. Among sites, the highest benthic diatom cell densities (4.6 × 105 cells cm−2) after 6 d was found in Lucero. Generic richness and diversity varied among sites, with the highest diversity (H’ = 1.58) on Day 1 in Tomasa, the site nearest to the mariculture area, whereas the highest richness (D = 2.17–3.26) for all days and diversity (H’ = 1.38–1.52) for Days 3 and 6 were recorded in Silaqui, the most distal site. Generally, the highest diversity and richness were found on Day 1, which then decreased on succeeding days in all sites. The lack of a clear patternin community structure among sites relative to the proximity of these sites to the mariculture area may in part be attributed to the presence of other nutrient sources. Results of the study provide baseline information on the variability of periphytic diatom community structure in a mariculture-impacted area, and insights on how benthic diatoms may be used in monitoring the impacts of nutrient pollution.This work was supported by the Department of Science and Technology–Philippine Council for Agriculture, Aquatic, and Natural Resources Research and Development under Grant QMSR-MRRDMEC-295-1449. The authors would like to thank Tirso O. Catbagan and Garry R. Bucol for their assistance during field sampling, and Timothy James Cipriano for the site map. We would also like to thank the Marine Biogeochemistry Laboratory of the Marine Science Institute, University of the Philippines Diliman for providing environmental data.
- Age, growth, and population structure of Conomurex luhunuasSanchez-Escalona, Katherine; Aliño, Porfirio (National Fisheries Research and Development Institute, 2022-12)Age and growth dynamics of Conomurex luhuanus were investigated to determine the population structure. Age-specific change in shell shape determined with geometric morphometrics revealed discrimination of shape between 0-3 years old at 99–100% while 3–4 years old can be separated with 81% certainty. Using the age discrimination data, K and L∞ were estimated at 1.00 year and 7.28 cm, respectively. Recruitment is bimodal with natural mortality (M) of 0.71 and fishing mortality (F) approximated at 3.92 year. The exploitation rate (E) is 0.85 year, indicating probable overharvesting of the population under study. The presence of a deep-water population, age-specific burying behavior, and bimodal recruitment pattern are possible resilience factors.The study was supported by a grant from PCAARRD-DOST.
- Associated effects of shading on the behavior, growth, and survival of Stichopus cf. horrens juvenilesRioja, Rose Angeli; Palomar-Abesamis, Nadia; Juinio–Meñez, Marie Antonette (Springer Science and Business Media LLC, 2021-07-07)Stichopus cf. horrens is an emergent culture species. It is known to be nocturnal and negatively phototactic; hence, determining its behavioral and growth responses to different light regimes is essential in optimizing culture protocols. This study examined the interactive effects of shading and relative food availability on the feeding pattern, absolute growth rates, and survival of juvenile S. cf. horrens. Six-month-old juveniles (4.27 to 19.41 g) were reared in replicate aquaria with three different shading treatments (covered, exposed, and half-covered aquaria) under ambient light conditions for 30 days. Juveniles in the exposed treatment with high microalgal biomass (13.44 ± 4.57 mg/g) had the highest growth rates (0.10 ± 0.05 g/day) but also the highest mortality (50%). In contrast, juveniles in the covered treatment had the lowest growth (−0.07 ± 0.03 g/day) but the highest survival (100%). Growth rates in the half-covered treatment were comparable with the exposed, and survival was higher than in the covered treatment. These indicate a trade-off between growth associated with more food and mortality risks due to light-induced stress. Juveniles in the half-covered treatment showed a significant preference to stay in the shaded portion whenever they were inactive during the day, suggesting avoidance to high light intensities (4726.51 ± 1582.43 Lux). Results of this study suggest that careful calibration of light intensities in nursery systems may help enhance juvenile growth. For indoor systems that may have limited space or surfaces for microalgal growth, cultured benthic diatom can be added to the partially covered tanks to increase the food available for the juveniles.
- Fish farm effluents alter reef benthic assemblages and reduce coral settlementQuimpo, Timothy Joseph R.; Ligson, Charlon A.; Manogan, Dana P.; Requilme, Jeremiah Noelle; Albelda, Ritzelle L.; Conaco, Cecilia; Cabaitan, Patrick C. (Elsevier Ltd., 2020)Fish farming in coastal areas is a rapidly growing industry. However, unregulated fish farming practices that release massive amounts of unconsumed feed and fecal material into the water column, can result in a nutrient enriched environment that extends to nearby reef systems. To understand the impact of fish farm effluent on coral settlement, we tested the settlement rate of Pocillopora acuta larvae on artificial substrates conditioned for 12 weeks at three sites with increasing distance (2–10 km) from fish farms in Bolinao, Philippines. Sites far from the fish farms had higher biofilm and crustose coralline algae cover. In contrast, the site closest to the fish farms, where nutrient levels were higher, had greater sediment and turf algae cover. Tiles conditioned at the farther sites promoted higher (6–8%) larval settlement whereas tiles from the nearer site had lower settlement (3%). These findings show that fish farm effluents can indirectly affect coral settlement on adjacent reefs by promoting growth of other biota that may inhibit larval settlement and by reducing the availability of suitable substrate.
- 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.
- Modified oxylipins as inhibitors of biofilm formation in Staphylococcus epidermidisPeran, Jacquelyn E.; Salvador-Reyes, Lilibeth A. (Frontiers Media SA, 2024-05-23)New approaches to combating microbial drug resistance are being sought, with the discovery of biofilm inhibitors considered as alternative arsenal for treating infections. Natural products have been at the forefront of antimicrobial discovery and serve as inspiration for the design of new antibiotics. We probed the potency, selectivity, and mechanism of anti-biofilm activity of modified oxylipins inspired by the marine natural product turneroic acid. Structure-activity relationship (SAR) evaluation revealed the importance of the trans-epoxide moiety, regardless of the position, for inhibiting biofilm formation. trans-12,13-epoxyoctadecanoic acid (1) and trans-9,10 epoxyoctadecanoic acid (4) selectively target the early stage of biofilm formation, with no effect on planktonic cells. These compounds interrupt the formation of a protective polysaccharide barrier by significantly upregulating the ica operon’s transcriptional repressor. This was corroborated by docking experiment with SarA and scanning electron micrographs showing reduced biofilm aggregates and the absence of thread-like structures of extrapolymeric substances. In silico evaluation revealed that 1 and 4 can interfere with the AgrA-mediated communication language in Staphylococci, typical to the diffusible signal factor (DSF) capacity of lipophilic chains.
- Sargassum sp. juice as an early juvenile supplemental feed for Stichopus cf. horrensIbañez, Glaiza; Cabanayan-Soy, Rona; Baure, Jerwin; Juinio-Meñez, Marie Antonette (Springer, 2022-09-28)The development of an efficient and low-cost feed is important to reduce the production and maintenance costs of microalgae. In this study, two experiments were conducted to evaluate the feasibility of using Sargassum sp. juice (SARG) to boost the growth and survival of post-settled Stichopus cf. horrens juveniles. Experiment 1 tested whether SARG improves growth compared with live microalgae diets, Chaetoceros calcitrans (Cc), combined Cc and Navicula ramosissima (Nr), and an unfed treatment. Experiment 2 determined the best SARG concentration—high feeding regime (HFR; 1 mL ind−1), medium (MFR; 0.5 mL ind−1), or low (LFR; 0.25 ml ind−1), relative to live microalgae Chaetoceros muelleri (CM). Juveniles in both experiments were reared for 30 days. In Experiment 1, the average daily growth rate (DGRL) of juveniles in SARG (0.04 ± 0.01 cm d−1) was the highest although not significantly different from Cc and Cc + Nr, but was significantly higher than the control. In Experiment 2, DGRL at day 14 in HFR (− 0.02 ± 0.02 cm d−1) was significantly lower than LFR (0.01 ± 0.01 cm d−1) and MFR (0.02 ± 0.02 cm d−1). Survival was higher in all SARG treatments compared with CM, while a significant decrease in feeding activity was observed in HFR by day 30. Results indicate that concentrations of 0.25–0.5 mL SARG per juvenile can boost growth and be an alternate diet for post-settled juveniles during early rearing. However, SARG alone is not sufficient to maintain growth beyond 3 weeks. With SARG feed supplementation and water quality management, the scaling-up of juvenile production of this emergent culture species can be accelerated.Our sincere gratitude to Mr. Tirso Catbagan and Mr. Garry Bucol for their assistance in the set-up of the experiments. We also thank Ms. Rose Angeli Rioja and Ms. JayR Gorospe for providing inputs to improve this paper. We also thank the Sea cucumber Research Team and the staff of the University of the Philippines—Marine Science Institute, Bolinao Marine Laboratory for their support and assistance during the conduct of the study.
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