National Committee on Marine Sciences (NCMS)
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- Microbiome diversity and host immune functions influence survivorship of sponge holobionts under future ocean conditionsPosadas, Niño; Baquiran, Jake Ivan P; Nada, Michael Angelou L; Kelly, Michelle; Conaco, Cecilia (Oxford University Press, 2021-07-03)The sponge-associated microbial community contributes to the overall health and adaptive capacity of the sponge holobiont. This community is regulated by the environment and the immune system of the host. However, little is known about the effect of environmental stress on the regulation of host immune functions and how this may, in turn, affect sponge–microbe interactions. In this study, we compared the bacterial diversity and immune repertoire of the demosponge, Neopetrosia compacta, and the calcareous sponge, Leucetta chagosensis, under varying levels of acidification and warming stress based on climate scenarios predicted for 2100. Neopetrosia compacta harbors a diverse microbial community and possesses a rich repertoire of scavenger receptors while L. chagosensis has a less diverse microbiome and an expanded range of pattern recognition receptors and immune response-related genes. Upon exposure to RCP 8.5 conditions, the microbiome composition and host transcriptome of N. compacta remained stable, which correlated with high survival (75%). In contrast, tissue necrosis and low survival (25%) of L. chagosensis was accompanied by microbial community shifts and downregulation of host immune-related pathways. Meta-analysis of microbiome diversity and immunological repertoire across poriferan classes further highlights the importance of host–microbe interactions in predicting the fate of sponges under future ocean conditions.We thank Francis Kenith Adolfo, Robert Valenzuela, and Ronald De Guzman for field and hatchery assistance and staff of the Bolinao Marine Laboratory for logistical support. This study was funded by the Department of Science and Technology Philippine Council for Agriculture, Aquatic and Natural Resources Research and Development (QMSR-MRRD-MEC-295-1449) to CC.
- Anti-inflammatory activity of monosubstituted xestoquinone analogues from the marine sponge Neopetrosia compactaSusana, Shalice R.; Salvador-Reyes, Lilibeth A. (MDPI, 2022-03-22)Chronic inflammation is recognized as a contributor to multiple chronic diseases, such as cancer, cardiovascular, and autoimmune disorders. Here, a natural products-initiated discovery of anti-inflammatory agents from marine sponges was undertaken. From the screening of 231 crude extracts, a total of 30 extracts showed anti-inflammatory activity with no direct cytotoxic effects at 50 μg/mL on RAW 264.7 (ATCC®TIB-71™) murine macrophage cells stimulated with 1 μg/mL lipopolysaccharide (LPS). Bioactivity-guided purification of the anti-inflammatory extract from the sponge Neopetrosia compacta led to the isolation of xestoquinone (1), adociaquinone B (2), adociaquinone A (3), 14-hydroxymethylxestoquinone (4), 15-hydroxymethylxestoquinone (5), and an inseparable 2:1 mixture of 14-methoxyxestoquinone and 15-methoxyxestoquinone (6). Compounds 1–6 caused a concentration-dependent reduction of nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells, with 4–6 having low micromolar IC50 and acceptable selectivity index. Gene expression analysis using qRT-PCR showed that 1, 5, and 6 downregulated Il1b and Nos2 expression by 2.1- to 14.8-fold relative to the solvent control at 10 μM. Xestoquinone (1) and monosubstituted analogues (4–6), but not the disubstituted adociaquinones (2 and 3), caused Nrf2 activation in a luciferase reporter MCF7 stable cells. Compounds 5 and 6 caused a modest increase in Nqo1 gene expression at 10 μM. The anti-inflammatory activity of xestoquinone (1) and monosubstituted analogues (4–6) may, in part, be mediated by Nrf2 activation, leading to attenuation of inflammatory mediators such as IL-1β and NOS2.Samples were collected under gratuitous permit numbers GP-0084-15 and GP-0123-17, issued by the Department of Agriculture of the Philippines. We thank the municipalities of Bolinao, Pangasinan, and Puerto Galera, Oriental Mindoro for permission for sample collection. We acknowledge assistance from Z. L. Malto and DDHP chemical ecology group in obtaining the mass spectrometric data and sample collection, respectively.
- Spatial variation in the benthic community structure of a coral reef system in the central Philippines: Highlighting hard coral, octocoral, and sponge assemblagesLalas, Jue Alef A.; Manzano, Geminne G.; Desabelle, Lee Arraby B.; Baria-Rodriguez, Maria Vanessa (Elsevier, 2023-07)Coral reefs are complex habitats that contain very high biodiversity and provide different ecosystem services. In the Coral Triangle, however, various major benthic components are still understudied. This can limit our understanding of coral reef community dynamics, especially in the presence of a changing climate coupled with local disturbances (e.g., decreased water quality). This study describes the benthic community structure of an ecologically and economically important coral reef system in the central Philippines through characterizing the assemblages of three major components (hard corals, octocorals, and sponges) among sites and stations with varying environmental conditions (i.e., exposure to monsoons, water quality levels). Results reveal significant variations in the mean percentage covers of hard corals, octocorals, and sponges at the site and station levels (ANOVA, p < 0.05), with hard corals dominating in Site 1, which is more exposed to the southwest monsoon, and Site 3, which is an embayed and unexposed site with low water quality, while soft corals dominated in Site 2, which is more exposed to the northeast monsoon. Multivariate analyses also revealed significant variations in the benthic community structure at different spatial scales (ANOSIM, p < 0.05). Interestingly, even stations within a site had significant variations in community structure, with different taxa being dominant. This study highlights the importance of conducting more detailed analyses of understudied taxa (i.e., octocorals and sponges) during coral reef surveys to improve our understanding of coral reef community dynamics that is very important for management.
- Genomics and metabolomics-based assessment of the biosynthetic potential of the sponge-associated microorganism Streptomyces cacaoi strain R2A-843A from the PhilippinesMalto, Zabrina Bernice L.; Reyes, Joeriggo M.; Lo, Bernard Isaiah; Davis, Kevin Bossie S.; Concepcion, Gisela; Salvador-Reyes, Lilibeth A. (Philippine-American Academy of Science and Engineering, 2023-10-20)The biosynthetic machinery of the sponge-associated Streptomyces cacaoi strain R2A-843A was assessed using a combined genomics and metabolomics approach. Whole genome sequencing and molecular networking showed the high biosynthetic potential of this actinomycete. A significant proportion of the genome is dedicated to secondary metabolite production, with biosynthetic gene clusters for nonribosomal peptides, polyketides, and terpenes being the most represented. Seven cyclic pentapeptides, including a putative new analogue, and a glycosylated lanthipeptide were identified using HRMS and untargeted MS/MS analysis. To validate our genome and metabolome analysis, we undertook a mass spectrometry-guided purification and confirmed the production of the known peptides BE-18257A (1) and BE-18257B (2). The production of 1 and 2 and the growth of the microorganism were monitored for eight days. Compound 2 was produced at a higher concentration, starting at 48 h post-incubation. Both compounds were noncytotoxic against colorectal and breast cancer cell lines.The authors acknowledge funding support from the Department of Science and Technology - Philippine Council for Health Research and Development through the Discovery and Development of Health Products - Marine Component Program. Genome sequencing was made possible through the CHEDPCARI IHITM63 Project. We thank Ms. Shalice R. SusanaGuevarra for conducting the bioactivity assay. This work was done under the supervision of the Bureau of Fisheries and Aquatic Resources under Gratuitous Permit No. FBP-0035-10. This is MSI Contribution No. 501.
- Genome mining of a novel marine sponge symbiont Nocardia sp. BML-15-R-026U reveals high biosynthetic potential for secondary metabolites, including a non-ribosomal peptide and a polyketide of high noveltyGloria, Paul Christian; Romines, Elaine; Punzalan, Marc Jeremie; Florece, Christine Marie; Cadorna, Kreighton; Salvador-Reyes, Lilibeth; Lluisma, Arturo (Philippine-American Academy of Science and Engineering, 2023-11-28)Antibiotic and drug resistance poses serious global public health threats, leading to substantial infections and fatalities annually. Addressing these issues requires the discovery of novel bioactive compounds and a faster and more cost-effective discovery process. However, traditional approaches, which require isolation and multi-step purification of compounds from organisms and running of initial assays, suffer from serious limitations such as the need for substantial amounts of biological material and high rates of compound rediscoveries. Because the biosynthetic capabilities of organisms are encoded in their genomes, genome mining provides a promising solution that would complement traditional approaches. This study conducted long-read whole genome sequencing on a marine sponge symbiont, Nocardia sp. BML-15-R-026U, to explore its genomic repertoire of secondary metabolite-encoding Biosynthetic Gene Clusters (BGCs). A four-contig genome assembly was generated for this isolate with a high degree of completeness and an estimated genome size of 4.84 Mbp. Its genome displays remarkable biosynthetic potential by containing at least 34 distinct secondary metabolite BGCs, predominantly Non-Ribosomal Peptide Synthetase (NRPS) and Polyketide Synthase (PKS) systems capable of producing novel chemical structures. Further analysis was focused on two genomic regions. In region 3.10, the study predicted a BGC for a novel, serine-rich non-ribosomal peptide with a predicted molecular weight of 2754 g/mol. Region 3.12 contained an iterative type-I PKS BGC, suggesting the potential synthesis of a polyketide compound with oxidoreductase-inhibiting properties. This study highlights genome mining as a productive early-phase approach for identifying promising drug leads and has identified the most promising candidates among this isolate’s BGCs for experimental validation.The study was funded by the Philippine Council for Health Research and Development – Department of Science and Technology under the “Anti-infective and Anticancer Drug Candidates from Marine Microorganisms and Sponges: Discovery and Development” project, Marine Science Institute – UP Diliman. The authors would like to thank the researchers of the Marine Genomics and Molecular Genetics Laboratory, MSI. The authors would also like to thank the researchers of the Discovery and Development of Health Products – Marine Component Phase I and researchers of the Marine Pharmacognosy Laboratory for the collection and initial analysis of the sample used in this study and storage and maintenance of the bacterial cultures. Sample collection was done under Gratuitous Permit No. GP-0084-15.
- Genistein and daidzein from a sponge-associated fungus (Family: Microstromataceae) show dose and incubation time-dependent Ca2+ influx activity variationAzcuna, Miguel Enrique Ma.; Acyatan, Zildjian; Manzano, Geminne; Yu, Clairecynth; Aliño, Porfirio Alexander M.; Altamia, Marvin; Salvador-Reyes, Lilibeth; Concepcion, Gisela P. (Philippine Association for the Advancement of Science and Technology, 2023-10-31)The compounds genistein and daidzein were obtained from the broth culture of a fungus isolated from the Philippine blue sponge Xestospongia sp. Genomic sequencing (18S rRNA) resulted in no exact hits and low sequence similarity (91%) to two species of fungi under the family Microstromataceae: Sympodiomycopsis vantaiensis and Microstromatales sp. Genistein has gained attention in recent years because of its potential to delay the onset of Alzheimer’s disease. This is the first report of genistein and daidzein isolated from a marine-derived fungus. Genistein and daidzein have a wide range of biological activities (e.g., neuroprotective, antimicrobial, anticancer), and this study reports a variation in intracellular [Ca2+] levels in dorsal root ganglion cells (DRGs) post-administration depending on dose and incubation time. An incubation time of 10 min resulted in a block effect, which was evidenced by decreased intracellular [Ca2+] levels. A dose-response was observed as the intensity of intracellular [Ca2+] decreased further at a higher dose. Conversely, an incubation time of 5 min resulted in an increase effect which was evidenced by decreased intracellular [Ca2+] levels. The similarity of these compounds with potent estrogens indicates that estrogen-mediated receptor signaling is the mechanism of action for the increase effect. The block effect, however, could be caused by a variety of factors, such as neurotoxicity or an ER stress response that results in the release of pro- and anti-apoptotic proteins. These findings confirm the ability of genistein to regulate [Ca2+] influx and the expression of apoptosis-related proteins. Further studies should investigate these mechanisms to understand the neuroprotective activities of genistein and daidzein.We thank the Drug Discovery and Health Products (DDHP) – Marine Component Project 1 for providing the sponge sample for fungal isolation. We thank Dr. Eizadora Yu of the Institute of Chemistry, University of the Philippines Diliman for providing fungal primers for DNA extraction and obtaining DNA sequences from the fungal sample.