The Harmful Unarmored Dinoflagellate Karlodinium in Japan and Philippines, with Reference to Ultrastructure and Micropredation of Karlodinium azanzae sp. nov. (Kareniaceae, Dinophyceae)1.

In all, 26 cultures of the harmful marine dinoflagellate Karlodinium, isolated from Japanese and Philippine coastal waters, were examined using LM, SEM, and molecular phylogeny inferred from ITS and LSU rDNA. Seven Karlodinium species (six from Japan and four from Philippines), K. australe, K. ballantinum, K. decipiens, K. gentienii, K. veneficum, K. zhouanum, and a novel species Karlodinium azanzae sp. nov., were identified based on their morphology and phylogenetic positions. Karlodinium azanzae from Manila Bay, Philippines was further characterized by TEM, HPLC (chloroplast pigment), and bioassay on brine shrimp and other marine zooplankton. Cells of K. azanzae were the largest (mean 25.3 µm long) in Karlodinium, possessed numerous tiny reflective particles, starch grains, and lipid granules, and usually swam at the bottom of the culture vessel. The straight apical structure complex and a ventral pore were common to the genus. The longitudinally elongated nucleus was located at the center, and the yellowish chloroplasts contained an embedded pyrenoid and carotenoid pigments typical of the genus (i.e., fucoxanthin as major carotenoid with its derivatives). TEM revealed a part of the flagellar apparatus, of which the long striated ventral connective is the first report in the Kareniaceae. Phylogenetic trees showed closest affinity of K. azanzae with K. australe and K. armiger. The new species could be differentiated from related species by cell size, position of the nucleus, and characteristic swimming behavior. Lethality of K. azanzae to large zooplankton and micropredation using a developed peduncle was also observed.

Development and application of a low-cost rapid assessment system for coastal benthic habitats.

Information on community structure and spatial distribution of benthic habitats are important in marine conservation and management. Coral areas, often monitored by trained scuba divers, are of importance for the ecosystem goods and services they provide. The data gathered may, however, be limited due to cost and time constraints. These restrictions and the continuing decline in coral reef health contribute to the need to develop rapid methods to efficiently document the distribution and status of coral reefs. A Rapid Assessment Instrument for Coastal Benthic Habitats (ARAICoBeH) System was developed to enable large-scale but low-cost coastal benthic habitat characterization and mapping without compromising accuracy. Inquiries on community structure and status in coral-dominated areas are also possible with the data collected using the method. The instrument, as well as the data processing and mapping algorithm are described in detail in this paper. A comparative study was conducted between ARAICoBeH and the frequently used underwater photo transect method. Variables compared were percent coral cover, functional group diversity, community structure as well as time and monetary requirements. There were no significant differences in estimates of percent coral cover and diversity of benthic functional groups for majority of sites while estimates of community structure were very similar. ARAICoBeH is cheaper and requires less time to gather the same amount of data as that of the photo transect method. The spatial distribution of coastal benthic habitats and community structure of coral-dominated areas in El Nido, Palawan, Philippines, are included to illustrate the application of the method.

Connections and clustering of Paralytic Shellfish Toxin events among coastal embayments in an archipelago partly mediated by advection.

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.

Thecal tabulation, body scale morphology and phylogeny of Heterocapsa philippinensis sp. nov. (Peridiniales, Dinophyceae) from the Philippines.

The thecal tabulation and body scale structure of the marine armoured dinoflagellate Heterocapsa, isolated from Philippines, were examined using LM, SEM and TEM, and its phylogenetic position was inferred from ITS and LSU rDNA sequences. Cells were ovoid and the plate tabulation (Po, cp, X, 5', 3a, 7'', 6c, 5s, 5''', 2'''') was consistent with most Heterocapsa species. The second anterior intercalary plate (2a) had a circular pattern with a thick marginal border free of pores. The nucleus was longitudinally elongated and curved, and located at the dorsal side of the cell. Discoid lobes of brownish chloroplast were peripherally distributed, and a pyrenoid was positioned at the centre. The triradiate body scales, measuring 250-300 nm in diameter, consisted of a roundish basal plate with six radiating ridges, nine peripheral uprights/spines, and three radiating spines. These components were identical to those of H. pseudotriquetra and H. steinii, except for the roundish outline of basal plate. Molecular phylogeny showed that the species clustered with H. pseudotriquetra and H. steinii. This species was differentiated from all other Heterocapsa species in the sausage-shaped nucleus and circular pattern on the 2a plate. This study proposed a novel species Heterocapsa philippinensis sp. nov. for the isolate.

Over 30 years of HABs in the Philippines and Malaysia: What have we learned?

In the Southeast Asian region, the Philippines and Malaysia are two of the most affected by Harmful Algal Blooms (HABs). Using long-term observations of HAB events, we determined if these are increasing in frequency and duration, and expanding across space in each country. Blooms of Paralytic Shellfish Toxin (PST)-producing species in the Philippines did increase in frequency and duration during the early to mid-1990s, but have stabilized since then. However, the number of sites affected by these blooms continue to expand though at a slower rate than in the 1990s. Furthermore, the type of HABs and causative species have diversified for both toxic blooms and fish kill events. In contrast, Malaysia showed no increasing trend in the frequency of toxic blooms over the past three decades since Pyrodinium bahamense was reported in 1976. However, similar to the Philippines, other PST producers such as Alexandrium minutum and Alexandrium tamiyavanichii have become a concern. No amnesic shellfish poisoning (ASP) has been confirmed in either Philippines or Malaysia thus far, while ciguatera fish poisoning cases are known from the Philippines and Malaysia but the causative organisms remain poorly studied. Since the 1990s and early 2000s, recognition of the distribution of other PST-producing species such as species of Alexandrium and Gymnodinium catenatum in Southeast Asia has grown, though there has been no significant expansion in the known distributions within the last decade. A major more recent problem in the two countries and for Southeast Asia in general are the frequent fish-killing algal blooms of various species such as Prorocentrum cordatum, Margalefidinium polykrikoides, Chattonella spp., and unarmored dinoflagellates (e.g., Karlodinium australe and Takayama sp.). These new sites affected and the increase in types of HABs and causative species could be attributed to various factors such as introduction through mariculture and eutrophication, and partly because of increased scientific awareness. These connections still need to be more concretely investigated. The link to the El Niño Southern Oscillation (ENSO) should also be better understood if we want to discern how climate change plays a role in these patterns of HAB occurrences.

Perceived global increase in algal blooms is attributable to intensified monitoring and emerging bloom impacts.

Global trends in the occurrence, toxicity and risk posed by harmful algal blooms to natural systems, human health and coastal economies are poorly constrained, but are widely thought to be increasing due to climate change and nutrient pollution. Here, we conduct a statistical analysis on a global dataset extracted from the Harmful Algae Event Database and Ocean Biodiversity Information System for the period 1985-2018 to investigate temporal trends in the frequency and distribution of marine harmful algal blooms. We find no uniform global trend in the number of harmful algal events and their distribution over time, once data were adjusted for regional variations in monitoring effort. Varying and contrasting regional trends were driven by differences in bloom species, type and emergent impacts. Our findings suggest that intensified monitoring efforts associated with increased aquaculture production are responsible for the perceived increase in harmful algae events and that there is no empirical support for broad statements regarding increasing global trends. Instead, trends need to be considered regionally and at the species level.

The harmful raphidophyte Chattonella (Raphidophyceae) in Western Pacific: Its red tides and associated fisheries damage over the past 50 years (1969-2019).

Red tides and associated fisheries damage caused by the harmful raphidophyte Chattonella were reassessed based on the documented local records for 50 years to understand the distribution and economic impacts of the harmful species in the Western Pacific. Blooms of Chattonella with fisheries damage have been recorded in East Asia since 1969, whereas they have been only recorded in Southeast Asia since the 1980s. Occurrences of Chattonella have been documented from six Southeast Asian countries, Indonesia, Malaysia, Philippines, Singapore, Thailand and Viet Nam, with mass mortalities mainly of farmed shrimp in 1980-1990s, and farmed fish in 2000-2010s. These occurrences have been reported with the names of C. antiqua, C. marina, C. ovata, C. subsalsa and Chattonella sp., owing to the difficulty of microscopic species identification, and many were not supported with molecular data. To determine the distribution of C. marina complex and C. subsalsa in Southeast Asia, molecular phylogeny and microscopic observation were also carried out for cultures obtained from Indonesia, Malaysia, Japan, Philippines, Russia, Singapore and Thailand. The results revealed that only the genotype of C. marina complex has been detected from East Asia (China, Japan, Korea and Russia), whereas both C. marina complex (Indonesia and Malaysia) and C. subsalsa (Philippines, Singapore and Thailand) were found in Southeast Asia. Ejection of mucocysts has been recognized as a diagnostic character of C. subsalsa, but it was also observed in our cultures of C. marina isolated from Indonesia, Malaysia, Japan, and Russia. Meanwhile, the co-occurrences of the two harmful Chattonella species in Southeast Asia, which are difficult to distinguish solely based on their morphology, suggest the importance of molecular identification of Chattonella genotypes for further understanding of their distribution and negative impacts.

Predicting fish kills and toxic blooms in an intensive mariculture site in the Philippines using a machine learning model.

Harmful algal blooms (HABs) that produce toxins and those that lead to fish kills are global problems that appear to be increasing in frequency and expanding in area. One way to help mitigate their impacts on people's health and livelihoods is to develop early-warning systems. Models to predict and manage HABs typically make use of complex multi-model structures incorporating satellite imagery and frequent monitoring data with different levels of detail into hydrodynamic models. These relatively more sophisticated methods are not necessarily applicable in countries like the Philippines. Empirical statistical models can be simpler alternatives that have also been successful for HAB forecasting of toxic blooms. Here, we present the use of the random forest, a machine learning algorithm, to develop an early-warning system for the prediction of two different types of HABs: fish kill and toxic bloom occurrences in Bolinao-Anda, Philippines, using data that can be obtained from in situ sensors. This site features intensive and extensive mariculture activities, as well as a long history of HABs. Data on temperature, salinity, dissolved oxygen, pH and chlorophyll from 2015 to 2017 were analyzed together with shellfish ban and fish kill occurrences. The random forest algorithm performed well: the fish kill and toxic bloom models were 96.1% and 97.8% accurate in predicting fish kill and shellfish ban occurrences, respectively. For both models, the most important predictive variable was a decrease in dissolved oxygen. Fish kills were more likely during higher salinity and temperature levels, whereas the toxic blooms occurred more at relatively lower salinity and higher chlorophyll conditions. This demonstrates a step towards integrating information from data that can be obtained through real-time sensors into a an early-warning system for two different types of HABs. Further testing of these models through times and different areas are recommended.

Larval connectivity patterns of the North Indo-West Pacific coral reefs.

Coral reefs of the North Indo-West Pacific provide important ecosystem services to the region but are subjected to multiple local and global threats. Strengthening management measures necessitate understanding the variability of larval connectivity and bridging global connectivity models to local scales. An individual-based Lagrangian biophysical model was used to simulate connectivity between coral reefs for three organisms with different early life history characteristics: a coral (Acropora millepora), a sea urchin (Tripneustes gratilla), and a reef fish (Epinephelus sp). Connectivity metrics and reef clusters were computed from the settlement probability matrices. Fitted power law functions derived from the dispersal kernels provided relative probabilities of connection given only the distance between reefs, and demonstrated that 95% of the larvae across organisms settled within a third of their maximum settlement distances. The magnitude of the connectivity metric values of reef cells were sensitive to differences both in the type of organism and temporal variability. Seasonal variability of connections was more dominant than interannual variability. However, despite these differences, the moderate to high correlation of metrics between organisms and seasonal matrices suggest that the spatial patterns are relatively similar between reefs. A cluster analysis based on the Bray-Curtis Dissimilarity of sink and source connections synthesized the inherent variability of these multiple large connectivity matrices. Through this, similarities in regional connectivity patterns were determined at various cluster sizes depending on the scale of interest. The validity of the model is supported by 1) the simulated dispersal kernels being within the range of reported parentage analysis estimates; and, 2) the clusters that emerged reflect the dispersal barriers implied by previously published population genetics studies. The tools presented here (dispersal kernels, temporal variability maps and reef clustering) can be used to include regional patterns of connectivity into the spatial management of coral reefs.

Paralytic Shellfish Toxin Uptake, Assimilation, Depuration, and Transformation in the Southeast Asian Green-Lipped Mussel (Perna viridis).

Bivalve molluscs represent an important food source within the Philippines, but the health of seafood consumers is compromised through the accumulation of harmful algal toxins in edible shellfish tissues. In order to assess the dynamics of toxin risk in shellfish, this study investigated the uptake, depuration, assimilation, and analogue changes of paralytic shellfish toxins in Perna viridis. Tank experiments were conducted where mussels were fed with the toxic dinoflagellate Alexandrium minutum. Water and shellfish were sampled over a six day period to determine toxin concentrations in the shellfish meat and water, as well as algal cell densities. The maximum summed toxin concentration determined was 367 µg STX eq./100 g shellfish tissue, more than six times higher than the regulatory action limit in the Philippines. Several uptake and depuration cycles were observed during the study, with the first observed within the first 24 h coinciding with high algal cell densities. Toxin burdens were assessed within different parts of the shellfish tissue, with the highest levels quantified in the mantle during the first 18 h period but shifting towards the gut thereafter. A comparison of toxin profile data evidenced the conversion of GTX1,4 in the source algae to the less potent GTX2,3 in the shellfish tissue. Overall, the study illustrated the temporal variability in Perna viridis toxin concentrations during a modelled algal bloom event, and the accumulation of toxin from the water even after toxic algae were removed.

Diversity and community structure of marine microbes around the Benham Rise underwater plateau, northeastern Philippines.

Microbes are central to the structuring and functioning of marine ecosystems. Given the remarkable diversity of the ocean microbiome, uncovering marine microbial taxa remains a fundamental challenge in microbial ecology. However, there has been little effort, thus far, to describe the diversity of marine microorganisms in the region of high marine biodiversity around the Philippines. Here, we present data on the taxonomic diversity of bacteria and archaea in Benham Rise, Philippines, Western Pacific Ocean, using 16S V4 rRNA gene sequencing. The major bacterial and archaeal phyla identified in the Benham Rise are Proteobacteria, Cyanobacteria, Actinobacteria, Bacteroidetes, Marinimicrobia, Thaumarchaeota and, Euryarchaeota. The upper mesopelagic layer exhibited greater microbial diversity and richness compared to surface waters. Vertical zonation of the microbial community is evident and may be attributed to physical stratification of the water column acting as a dispersal barrier. Canonical Correspondence Analysis (CCA) recapitulated previously known associations of taxa and physicochemical parameters in the environment, such as the association of oligotrophic clades with low nutrient surface water and deep water clades that have the capacity to oxidize ammonia or nitrite at the upper mesopelagic layer. These findings provide foundational information on the diversity of marine microbes in Philippine waters. Further studies are warranted to gain a more comprehensive picture of microbial diversity within the region.

Insights into the dynamics of harmful algal blooms in a tropical estuary through an integrated hydrodynamic-Pyrodinium-shellfish model.

In contrast to temperate Harmful Algal Blooms (HABs), knowledge on the mechanisms driving tropical HABs are less well studied. The interaction of a seasonal temperature window, cysts (for certain species) and large-scale transport are some of the key processes in temperate HABs. In the Philippines, HABs occur not along long open coastlines, but in embayments that are highly influenced by run-off and stratification. These embayments are typically also the sites of cultured or wild harvest shellfish and other aquaculture activities. Sorsogon Bay in the northeastern Philippines has experienced prolonged shellfish-harvesting bans due to blooms by Pyrodinium bahamense var. compressum severely affecting the fisheries industry in this area, as well as leading to Paralytic Shellfish Poisoning illnesses and fatalities. A novel integrated model was developed that mechanistically captures the interactions between hydrodynamic conditions, nutrients, the life history (cells and cysts) of Pyrodinium, as well as the cultured shellfish within the bay and their ensuing toxicities due to ingestion of toxic Pyrodinium cells and cysts. This is the second model developed for HABs in the Philippines, and the first to integrate different components of Pyrodinium bloom dynamics. The model is modularly composed of a watershed nutrient and diffusion model, a 3D hydrodynamic model, a Pyrodinium population model and a shellfish toxin model. It was able to capture the observed temporal variations of Pyrodinium and shellfish toxicity. It was also able to represent some aspects of the spatial distribution in Sorsogon Bay though there were discrepancies. To explore the dynamics of blooms, the linkages between the bloom and decline of the Pyrodinium population with shellfish toxicity as affected by temperature, salinity and nutrients were investigated. Comparisons with field results showed the seasonality of blooms in Sorsogon Bay is driven by increased rainfall. The timing of these conditions is important in facilitating Pyrodinium excystment and reproduction. Model results showed as well the potential significance of shellfish grazing and dinoflagellate cell mortality in influencing the decline of the bloom, and toxicity levels. This approach is promising in helping to understand mechanisms for HABs more holistically, and the model can be further improved to provide more precise quantitative information.