Character evolution of the benthic thecate dinoflagellate, gambierdiscus (Dinophyceae), with an introduction of the interactive key to species

The genus Gambierdiscus is one of the benthic marine dinoflagellates that are known to produce biotoxins, causing ciguatera food poisoning (CFP) in humans after consuming the contaminated marine fishes. These benthic marine dinoflagellates are commonly found in the warm tropical and subtropical waters. The phenomena associated with high cell abundance or “blooms” is referred to Benthic Harmful Algal Blooms (BHABs). Precise species identification is crucial as not all of the species in the genus are toxic. In this study, morphological characteristics of Gambierdiscus species widely used in species identification were analyzed, and the character states coded. Taxon sampling on the large subunit (LSU) rDNA of all Gambierdiscus species were carried out, and used for the phylogenetic reconstruction. The character states were mapped onto the Maximum Parsimony (MP) tree to investigate the character state evolution of Gambierdiscus species. Morphological information and the distribution of the 13 species were used to develop a comprehensive taxonomic database of Gambierdiscus, a web-based interactive identification key for species identification is presented

Evolutionary lineages and character state evolution of the benthic thecated dinoflagellate, gambierdiscus (dinophyceae)

The genera of Gambierdiscus, Ostreopsis and Coolia are benthic marine dinoflagellates that are known to produce biotoxins that cause illness in human after consumption of the contaminated seafood. These benthic marine dinoflagellates are commonly found in the warm tropical and subtropical waters. The phenomena, of which these organisms bloom, are referred to Benthic Harmful Algal Bloom (BHAB). In this study, the morphology and genetic information of species in the genus Gambierdiscus were investigated. Samples were collected from Talang-Satang National Park, and single cells were isolated and cultures established in ES-DK medium. The cultured samples were undergone morphological observations by using light and scanning electron microscopes. Total genomic DNA was isolated, and the nuclear-encoded ribosomal RNA genes amplified and sequenced. The sequences obtained were analyzed and used for phylogenetic reconstruction to examine the phylogenetic lineage. The morphological characters used in species identification were coded and character matrix constructed based on data obtained in this study as well existing data from literature. The character states were mapped onto the phylogenetic tree to investigate the character state evolution of these species. The major outcome of this study is knowledge obtained for the phylogenetic relationship of the genus and findings of taxonomic-informative and phylogenetically significant morphological characters, which could be useful in aiding species identification of the species

Growth and epiphytic behavior of three Gambierdiscus species (Dinophyceae) associated with various macroalgal substrates

Species of the benthic dinoflagellate Gambierdiscus produce polyether neurotoxins that caused ciguatera fish/shellfish poisoning in human. The toxins enter marine food webs by foraging of herbivores on the biotic substrates like macroalgae that host the toxic dinoflagellates. Interaction of Gambierdiscus and their macroalgal substrate hosts is believed to shape the tendency of substrate preferences and habitat specialization. This was supported by studies that manifested epiphytic preferences and behaviors in Gambierdiscus species toward different macroalgal hosts. To further examine the supposition, a laboratory-based experimental study was conducted to examine the growth, epiphytic behaviors and host preferences of three Gambierdiscus species towards four macroalgal hosts over a culture period of 40 days. The dinoflagellates Gambierdiscus balechii, G. caribaeus, and a new ribotype, herein designated as Gambierdiscus type 7 were initially identified based on the thecal morphology and molecular characterization. Our results showed that Gambierdiscus species tested in this study exhibited higher growth rates in the presence of macroalgal hosts. Growth responses and attachment behaviors, however, differed among different species and strains of Gambierdiscus over different macroalgal substrate hosts. Cells of Gambierdiscus mostly attached to substrate hosts at the beginning of the experiments but detached at the later time. Localized Gambierdiscus-host interactions, as demonstrated in this study, could help to better inform efforts of sampling and monitoring of this benthic toxic dinoflagellate

Habitat complexity affects benthic harmful dinoflagellate assemblages in the fringing reef of Rawa Island, Malaysia

Few studies have investigated the effect of fine-scale habitat differences on the dynamics of benthic harmful dinoflagellate assemblages. To determine how these microhabitat differences affect the distribution and abundance of the major benthic harmful dinoflagellate genera in a tropical coral reef ecosystem, a field study was undertaken between April–September 2015 and January 2016 on the shallow reef flat of the fringing reef of Rawa Island, Terengganu, Malaysia. Sampling of benthic dinoflagellates was carried out using an artificial substrate sampling method (fiberglass screens). Benthic microhabitats surrounding the sampling screens were characterized simultaneously from photographs of a 0.25-m2 quadrat based on categories of bottom substrate types. Five taxonomic groups of benthic dinoflagellates, Ostreopsis, Gambierdiscus, Prorocentrum, Amphidinium, and Coolia were identified, and cells were enumerated using a light microscope. The results showed Gambierdiscus was less abundant than other genera throughout the study period, with maximum abundance of 1.2 × 103 cells 100 cm−2. While most taxa were present on reefs with high coral cover, higher cell abundances were observed in reefs with high turf algal cover and coral rubble, with the exception of Ostreopsis, where the abundance reached a maximum of 3.4 × 104 cells 100 cm−2 in habitats with high coral cover. Microhabitat heterogeneity was identified as a key factor governing the benthic harmful dinoflagellate assemblages and may account for much of the observed variability in dominant taxa. This finding has significant implications for the role of variability in the benthic harmful algal bloom (BHAB) outbreaks and the potential in identifying BHAB-related toxin transfer pathways and the key vectors in the food webs

Diversity of Heterocapsa (Dinophyceae) and the algal bloom event in the mariculture areas of Johor Strait, Malaysia

In November 2020, a high biomass multi-species algal bloom caused heavy water discoloration in the fish and mussel farm areas of the Johor Strait, Malaysia. A total of 19 microalgal taxa were identified from the plankton samples collected during the bloom event. Eleven genera were diatoms, and eight genera were dinophytes. The microalgal composition was dominated by the diatom Guinardia sp., with an average cell density of 1.7×106 cells L−1 , making up 65–80% of the phytoplankton composition. Concomitantly, high densities of the dinophytes Heterocapsa minima (3.8–5.3×105 cells L−1 ) and Karlodinium spp. (3.5–6.6×103 cells L−1 ) were found. This is the first record of the occurrence of H. minima in Malaysian waters. Detailed morphological observations of H. minima based on scanning electron microscopy are presented in this study. To have a better insight into the Heterocapsa species assemblages in the Johor Strait, the diversity of Heterocapsa species assemblages along the strait was investigated based on a metabarcoding approach. Environmental DNA collected between 2018 and 2019 was used for high throughput amplicon sequencing targeting the small subunit (SSU) ribosomal RNA gene marker. The metabarcoding analysis detected three rare Heterocapsa species in the waters, H. niei, H. rotundata, and H. steinii. The results showed that Heterocapsa species assemblages varied temporally across the strait, with higher species diversity and amplicon sequence variant (ASV) read abundances detected in the Eastern Johor Strait. Although no fish/shellfish kills were sighted during the 2020 bloom event, the presence of harmful microalgal species, such as Heterocapsa minima and Karlodinium spp., urged the need for a comprehensive HAB monitoring program in the Strait to safeguard the aquaculture industry in the areas