Ji Eun Kwon

Mixotrophic ability of the phototrophic dinoflagellates Alexandrium andersonii, A. affine, and A. fraterculus

The dinoflagellate Alexandrium spp. have received much attention due to their harmful effects on diverse marine organisms, including commercially important species. For minimizing loss due to red tides or blooms of Alexandrium spp., it is very important to understand the eco-physiology of each Alexandrium species and to predict its population dynamics. Its trophic mode (i.e., exclusively autotrophic or mixotrophic) is one of the most critical parameters in establishing prediction models. However, among the 35 Alexandrium species so far described, only six Alexandrium species have been revealed to be mixotrophic. Thus, mixotrophic ability of the other Alexandrium species should be explored. In the present study, whether each of three Alexandrium species (A. andersonii, A. affine, and A. fraterculus) isolated from Korean waters has or lacks mixotrophic ability, was investigated. When diets of diverse algal prey, cyanobacteria, and bacteria sized micro-beads were provided, A. andersonii was able to feed on the prasinophyte Pyramimonas sp., the cryptophyte Teleaulax sp., and the dinoflagellate Heterocapsa rotundata, whereas neither A. affine nor A. fraterculus fed on any prey item. Moreover, mixotrophy elevated the growth rate of A. andersonii. The maximum mixotrophic growth rates of A. andersonii on Pyramimonas sp. under a 14:10h light/dark cycle of 20μEm-2s-1 was 0.432d-1, while the autotrophic growth rate was 0.243d-1. With increasing mean prey concentration, the ingestion rate of A. andersonii increased rapidly at prey concentrations <650ngCml-1 (ca. 16,240 cellsml-1), but became saturated at the higher prey concentrations. The maximum ingestion rate by A. andersonii of Pyramimonas sp. was 1.03ngC predator-1d-1 (25.6 cells predator-1d-1). This evidence suggests that the mixotrophic ability of A. andersonii should be taken into consideration in predicting the outbreak, persistence, and decline of its harmful algal blooms.

Mixotrophy in the phototrophic dinoflagellate Takayama helix (family Kareniaceae): Predator of diverse toxic and harmful dinoflagellates

Takayama spp. are phototrophic dinoflagellates belonging to the family Kareniaceae and have caused fish kills in several countries. Understanding their trophic mode and interactions with co-occurring phytoplankton species are critical steps in comprehending their ecological roles in marine ecosystems, bloom dynamics, and dinoflagellate evolution. To investigate the trophic mode and interactions of Takayama spp., the ability of Takayama helix to feed on diverse algal species was examined, and the mechanisms of prey ingestion were determined. Furthermore, growth and ingestion rates of T. helix feeding on the dinoflagellates Alexandrium lusitanicum and Alexandrium tamarense, which are two optimal prey items, were determined as a function of prey concentration. T. helix ingested large dinoflagellates ≥15μm in size, except for the dinoflagellates Karenia mikimotoi, Akashiwo sanguinea, and Prorocentrum micans (i.e., it fed on Alexandrium minutum, A. lusitanicum, A. tamarense, A. pacificum, A. insuetum, Cochlodinium polykrikoides, Coolia canariensis, Coolia malayensis, Gambierdiscus caribaeus, Gymnodinium aureolum, Gymnodinium catenatum, Gymnodinium instriatum, Heterocapsa triquetra, Lingulodinium polyedrum, and Scrippsiella trochoidea). All these edible prey items are dinoflagellates that have diverse eco-physiology such as toxic and non-toxic, single and chain forming, and planktonic and benthic forms. However, T. helix did not feed on small flagellates and dinoflagellates <13μm in size (i.e., the prymnesiophyte Isochrysis galbana; the cryptophytes Teleaulax sp., Storeatula major, and Rhodomonas salina; the raphidophyte Heterosigma akashiwo; the dinoflagellates Heterocapsa rotundata, Amphidinium carterae, Prorocentrum minimum; or the small diatom Skeletonema costatum). T. helix ingested Heterocapsa triquetra by direct engulfment, but sucked materials from the rest of the edible prey species through the intercingular region of the sulcus. With increasing mean prey concentration, the specific growth rates of T. helix on A. lusitanicum and A. tamarense increased continuously before saturating at prey concentrations of 336-620ngC mL-1. The maximum specific growth rates (mixotrophic growth) of T. helix on A. lusitanicum and A. tamarense were 0.272 and 0.268d-1, respectively, at 20°C under a 14:10 h light/dark cycle of 20μE m-2 s-1 illumination, while its growth rates (phototrophic growth) under the same light conditions without added prey were 0.152 and 0.094d-1, respectively. The maximum ingestion rates of T. helix on A. lusitanicum and A. tamarense were 1.23 and 0.48ng C predator-1d-1, respectively. The results of the present study suggest that T. helix is a mixotrophic dinoflagellate that is able to feed on a diverse range of toxic species and, thus, its mixotrophic ability should be considered when studying red tide dynamics, food webs, and dinoflagellate evolution.

Differential interactions between the nematocyst-bearing mixotrophic dinoflagellate Paragymnodinium shiwhaense and common heterotrophic protists and copepods: Killer or prey

To investigate interactions between the nematocyst-bearing mixotrophic dinoflagellate Paragymnodinium shiwhaense and different heterotrophic protist and copepod species, feeding by common heterotrophic dinoflagellates (Oxyrrhis marina and Gyrodinium dominans), naked ciliates (Strobilidium sp. approximately 35μm in cell length and Strombidinopsis sp. approximately 100μm in cell length), and calanoid copepods Acartia spp. (A. hongi and A. omorii) on P. shiwhaense was explored. In addition, the feeding activities of P. shiwhaense on these heterotrophic protists were investigated. Furthermore, the growth and ingestion rates of O. marina, G. dominans, Strobilidium sp., Strombidinopsis sp., and Acartia spp. as a function of P. shiwhaense concentration were measured. O. marina, G. dominans, and Strombidinopsis sp. were able to feed on P. shiwhaense, but Strobilidium sp. was not. However, the growth rates of O. marina, G. dominans, Strobilidium sp., and Strombidinopsis sp. feeding on P. shiwhaense were very low or negative at almost all concentrations of P. shiwhaense. P. shiwhaense frequently fed on O. marina and Strobilidium sp., but did not feed on Strombidinopsis sp. and G. dominans. G. dominans cells swelled and became dead when incubated with filtrate from the experimental bottles (G. dominans+P. shiwhaense) that had been incubated for one day. The ingestion rates of O. marina, G. dominans, and Strobilidium sp. on P. shiwhaense were almost zero at all P. shiwhaense concentrations, while those of Strombidinopsis sp. increased with prey concentration. The maximum ingestion rate of Strombidinopsis sp. on P. shiwhaense was 5.3ngC predator-1d-1 (41 cells predator-1d-1), which was much lower than ingestion rates reported in the literature for other mixotrophic dinoflagellate prey species. With increasing prey concentrations, the ingestion rates of Acartia spp. on P. shiwhaense increased up to 930ngCml-1 (7180cellsml-1) at the highest prey concentration. The highest ingestion rate of Acartia spp. on P. shiwhaense was 4240ngC predator-1d-1 (32,610 cells predator-1d-1), which is comparable to ingestion rates from previous studies on other dinoflagellate prey species calculated at similar prey concentrations. Thus, P. shiwhaense might play diverse ecological roles in marine planktonic communities by having an advantage over competing phytoplankton in anti-predation against potential protistan grazers.

Mixotrophy in the newly described dinoflagellate Yihiella yeosuensis: A small, fast dinoflagellate predator that grows mixotrophically, but not autotrophically

To investigate tropical roles of the newly described Yihiella yeosuensis (ca. 8μm in cell size), one of the smallest phototrophic dinoflagellates in marine ecosystems, its trophic mode and the types of prey species that Y. yeosuensis can feed upon were explored. Growth and ingestion rates of Y. yeosuensis on its optimal prey, Pyramimonas sp. (Prasinophyceae), as a function of prey concentration were measured. Additionally, growth and ingestion rates of Y. yeosuensis on the other edible prey, Teleaulax sp. (Cryptophyceae), were also determined for a single prey concentration at which both these rates of Y. yeosuensis on Pyramimonas sp. were saturated. Among bacteria and diverse algal prey tested, Y. yeosuensis fed only on small Pyramimonas sp. and Teleaulax sp. (both cell sizes=5.6μm). With increasing mean prey concentrations, both specific growth and ingestion rates of Y. yeosuensis increased rapidly before saturating at a mean Pyramimonas concentration of 109ngCmL-1 (2725cellsmL-1). The maximum growth rate (mixotrophic growth) of Y. yeosuensis fed with Pyramimonas sp. at 20°C under a 14:10-h light-dark cycle of 20μEm-2s-1 was 1.32d-1, whereas the growth rate of Y. yeosuensis without added prey was 0.026d-1. The maximum ingestion rate of Y. yeosuensis fed with Pyramimonas sp. was 0.37ngCpredator-1d-1 (9.3cellspredator-1d-1). At a Teleaulax concentration of 1130ngCmL-1 (66,240cellsmL-1), growth and ingestion rates of Y. yeosuensis fed with Teleaulax sp. were 1.285d-1 and 0.38ngCpredator-1d-1 (22.4cellspredator-1d-1), respectively. Thus, Y. yeosuensis rarely grows without mixotrophy, and mixotrophy supports high growth rates in Y. yeosuensis. Y. yeosuensis has the highest maximum mixotrophic growth rate with the exception of Ansanella graniferaamong engulfment feeding mixotrophic dinoflagellates. However, the high swimming speed of Y. yeosuensis (1572μms-1), almost the highest among phototrophic dinoflagellates, may prevent autotrophic growth. This evidence suggests that Y. yeosuensis may be an effective mixotrophic dinoflagellate predator on Pyramimonas and Teleaulax, and occurs abundantly during or after blooms of these two prey species.

Removal of two pathogenic scuticociliates Miamiensis avidus and Miamiensis sp. using cells or culture filtrates of the dinoflagellate Alexandrium andersonii

Scuticociliatosis, which is caused by parasitic protistan pathogens known as scuticociliates, is one of the most serious diseases in marine aquaculture worldwide. Thus, elimination of these ciliates is a primary concern for scientists and managers in the aquaculture industry. To date, formalin and other toxic chemicals have been used as anti-scuticociliate agents, but issues regarding their secondary effects often arise. Consequently, development of safer methods is necessary. To find out a safe method of controlling scuticociliate populations in aqua-tanks or small-scale natural environments, cultures of 14 phototrophic dinoflagellates were tested to determine whether they were able to control populations of the common scuticociliates Miamiensis avidus and Miamiensis sp. isolated from Korean waters. Among the dinoflagellates tested, both cells and culture filtrates of Alexandrium andersonii effectively killed M. avidus and Miamiensis sp. The minimal concentration of cells and equivalent culture filtrates of A. andersonii to kill all M. avidus cells within 48h of incubation was ca. 2500 and 4500 cells ml-1, respectively; whereas those needed to kill all Miamiensis sp. cells were ca. 1000 and 4500 cells ml-1, respectively. It was estimated that 1m3 of the stock culture containing 20,000A. andersonii cells ml-1 could eliminate all M. avidus cells in 7m3 of waters within the aqua-tanks on land and all Miamiensis sp. cells in 19m3 of waters within 48h. None of the brine shrimp Artemia salina nauplii incubated with concentrations of 50-4500A. andersonii cells ml-1 for 24h was dead. Furthermore, none of the flounder Paralichthys olivaceus juveniles incubated with a mean concentration of ca. 2280A. andersonii cells ml-1 for 96h was dead. Therefore, A. andersonii cultures may be used as a safe biological method for controlling populations of scuticociliates and can replace toxic formalin. The results of this study provided the basis for developing the method to control scuticociliate populations and understanding interactions between scuticociliates and phototrophic dinoflagellates in marine ecosystems.

Newly discovered role of the heterotrophic nanoflagellate Katablepharis japonica, a predator of toxic or harmful dinoflagellates and raphidophytes

Heterotrophic nanoflagellates are ubiquitous and known to be major predators of bacteria. The feeding of free-living heterotrophic nanoflagellates on phytoplankton is poorly understood, although these two components usually co-exist. To investigate the feeding and ecological roles of major heterotrophic nanoflagellates Katablepharis spp., the feeding ability of Katablepharis japonica on bacteria and phytoplankton species and the type of the prey that K. japonica can feed on were explored. Furthermore, the growth and ingestion rates of K. japonica on the dinoflagellate Akashiwo sanguinea-a suitable algal prey item-heterotrophic bacteria, and the cyanobacteria Synechococcus sp., as a function of prey concentration were determined. Among the prey tested, K. japonica ingested heterotrophic bacteria, Synechococcus sp., the prasinophyte Pyramimonas sp., the cryptophytes Rhodomonas salina and Teleaulax sp., the raphidophytes Heterosigma akashiwo and Chattonella ovata, the dinoflagellates Heterocapsa rotundata, Amphidinium carterae, Prorocentrum donghaiense, Alexandrium minutum, Cochlodinium polykrikoides, Gymnodinium catenatum, A. sanguinea, Coolia malayensis, and the ciliate Mesodinium rubrum, however, it did not feed on the dinoflagellates Alexandrium catenella, Gambierdiscus caribaeus, Heterocapsa triquetra, Lingulodinium polyedra, Prorocentrum cordatum, P. micans, and Scrippsiella acuminata and the diatom Skeletonema costatum. Many K. japonica cells attacked and ingested a prey cell together after pecking and rupturing the surface of the prey cell and then uptaking the materials that emerged from the ruptured cell surface. Cells of A. sanguinea supported positive growth of K. japonica, but neither heterotrophic bacteria nor Synechococcus sp. supported growth. The maximum specific growth rate of K. japonica on A. sanguinea was 1.01 d-1. In addition, the maximum ingestion rate of K. japonica for A. sanguinea was 0.13ngC predator-1d-1 (0.06 cells predator-1d-1). The maximum ingestion rate of K. japonica for heterotrophic bacteria was 0.019ngC predator-1d-1 (266 bacteria predator-1d-1), and the highest ingestion rate of K. japonica for Synechococcus sp. at the given prey concentrations of up to ca. 107 cells ml-1 was 0.01ngC predator-1d-1 (48 Synechococcus predator-1d-1). The maximum daily carbon acquisition from A. sanguinea, heterotrophic bacteria, and Synechococcus sp. were 307, 43, and 22%, respectively, of the body carbon of the predator. Thus, low ingestion rates of K. japonica on heterotrophic bacteria and Synechococcus sp. may be responsible for the lack of growth. The results of the present study clearly show that K. japonica is a predator of diverse phytoplankton, including toxic or harmful algae, and may also affect the dynamics of red tides caused by these prey species.

Gonyaulax whaseongensis sp. nov. (Gonyaulacales, Dinophyceae), a new phototrophic species from Korean coastal waters

The planktonic phototrophic dinoflagellate Gonyaulax whaseongensis sp. nov., isolated from coastal waters of western Korea, was described from living and fixed cells under light and scanning electron microscopy, and its rDNA was sequenced. Gonyaulax whaseongensis had a plate formula of 2pr, 4′, 6′′, 6c, 6′′′, 1p, and 1′′′′ with S‐type ventral organization like the other species in the genus. However, this dinoflagellate had a narrow cingulum (ca. 2.6 μm), small displacement of the cingulum, slight overhang and steep angle between the ends of the cingulum, quadrangular sixth precingular plate, reticulated cell surface without longitudinal lines or ridges, and two unequal antapical spines, together which distinguish this from all other reported Gonyaulax species. In addition, the SSU and LSU rDNA sequences were 8%–12% and 11%–24%, respectively, different from those of Gonyaulax polygramma, Gonyaulax spinifera, Gonyaulax fragilis, Gonyaulax membranacea, and Gonyaulax digitale, the putatively closest related species in the phylogenetic analysis.