Occurrence ofDinophysis spp. and toxic shellfish in the Northern Adriatic

The dynamics of the toxicity of the musselMytilus galloprovincialis was compared between two different shellfish farms, 5 km apart, but using the same cultivation technique. The main differences concerned the freshwater influx and the open aspect to the Gulf of Trieste. It is suggested that a deep closed bay and abundant fresh water inflow are the two main conditions for the low toxicity levels in mussels and for shorter periods of danger. A detailed study of the phytoplankton samples revealed the presence of eight species ofDinophysis in the area of both shellfish farms. During the period of the DSP outbreak in Slovenia (autumn and winter 1989).D. fortii andD. acuminata were the most frequentDinophysis species. There was a high positive correlation between the onset of mussel toxicity and the appearance ofDinophysis spp.     

Phytoplankton composition of the Kandalaksha Gulf, Russian White Sea: Dinophysis and lipophilic toxins in the blue mussel (Mytilus edulis)

Dinophysis acuminata and D. norvegica were observed in plankton net samples during the summer of 2002 from the Kandalaksha Gulf in the White Sea (North European Russia). Prorocentrum lima was found as an epiphyte on subtidal macroalgae in August, but not observed in plankton net samples. Protein phosphatase 2A (PP2A) inhibition measured 127.8 ng OA-equivalent/g of mussel (Mytilus edulis) hepatopancreas from samples collected a few days after when Dinophysis was recorded at a density of 1550 cells L⁻¹. Liquid chromatography–mass spectrometry confirmed presence of several classes of lipophilic shellfish toxins associated with Dinophysis spp. in the mussels including okadaic acid, dinophysistoxin-1, pectenotoxins and yessotoxins. No azaspiracid was detected. This represents the first identification of phycotoxicity in the White Sea.     

Large subunit ribosomal RNA gene variation and sequence heterogeneity of Dinophysis (Dinophyceae) species from Scottish coastal waters

The purpose of the study was to investigate the genetic diversity of Dinophysis species from around the Scottish coast, with a view to an improved understanding of the dynamics and identification of this genus in Scottish waters. Single-cell PCR amplification with direct sequencing was performed on a total of 441 Dinophysis cells isolated from both live and Lugol's fixed plankton net samples. Universal eukaryotic primers were used to amplify the large subunit (LSU) ribosomal RNA (rRNA) gene of the Dinophysis isolates, with a frequency of PCR success of 26% for non-fixed and 48% for fixed samples. From this a total of 30 isolates were selected for this study and the D1–D2 region of the LSU-rRNA gene sequenced for phylogenetic analysis. No significant correlation could be made between geographical location and LSU sequence, although some regional sequence heterogeneity was observed within the Dinophysis acuta species. LSU sequence data was used to design Dinophysis genus specific and Dinophysis clade-specific primers primarily to ensure clean sequences from universal D1–D2 amplicons without a requirement for cloning. Three clade-specific primers designed to a region within the D2 hypervariable region of the LSU-rRNA gene allowed discrimination of Dinophysis acuminata/norvegica from Dinophysis tripos/caudata and Dinophysis fortii/acuta. In two isolates, SC359 (D. tripos) and LC58 (D. acuta), nested PCR products were observed with both the expected clade-specific primer, and Dasd-R2, the D. acuminata/norvegica clade-specific primer. Cloning and sequence analysis suggested that these amplicons were genuine “D. acuminata-like” sequences and their presence, albeit at a low frequency within different Dinophysis species, indicated that individual Dinophysis cells possess heterologous copies of the LSU-rRNA gene that are similar to LSU sequences normally associated with D. acuminata. The nature of the process that generated these hybrid cells, the frequency of such events and their importance is as yet unknown, but may provide a cautionary note for the development of PCR-based species specific detection methods.     

DSP toxin production de novo in cultures of Dinophysis acuminata (Dinophyceae) from North America

For decades, many aspects of Dinophysis biology have remained intractable due to our inability to maintain these organisms in laboratory cultures. Recent breakthroughs in culture methods have opened the door for detailed investigations of these important algae. Here, for the first time, we demonstrate toxin production in cultures of North American Dinophysis acuminata, isolated from Woods Hole, MA. These findings show that, despite the rarity of Dinophysis-related DSP events in North America, D. acuminata from this area has the ability to produce DSP toxins just as it does in other parts of the world where this species is a major cause of DSP toxicity. In our cultures, D. acuminata cells were observed feeding on Myrionecta rubra using a peduncle. Culture extracts were analyzed using LC–MS/MS, providing unequivocal evidence for the toxin DTX1 in the Dinophysis cultures. In addition, a significant amount of an okadaic acid diol ester, OA-D8, was detected. These results suggest that this Dinophysis isolate stores much of its OA as a diol ester. Also, toxin PTX-2 and a hydroxylated PTX-2 with identical fragmentation mass spectrum to that of PTX-11, but with a different retention time, were detected in this D. acuminata culture. This demonstration of toxin production in cultured North American Dinophysis sets the stage for more detailed studies investigating the causes of geographic differences in toxicity. It is now clear that North American Dinophysis have the ability to produce DSP toxins even though they only rarely cause toxic DSP events in nature. This may reflect environmental conditions that might induce or repress toxin production, genetic differences that cause modifications in toxin gene expression, or physiological and biochemical differences in prey species.     

Food selectivity and grazing impact on toxic Dinophysis spp. by copepods feeding on natural plankton assemblages

Food selectivity and grazing impact by Acartia bifilosa, Temora longicornis and Centropages typicus on Dinophysis spp. plankton assemblages were experimentally investigated in the Baltic Sea. Toxin analyses were carried out on phyto- and zooplankton-dominated size fractions from field-collected samples to assess if toxins produced by Dinophysis spp. would end up in the zooplankton. All copepod species fed actively on toxic Dinophysis spp. (Dinophysis acuta and Dinophysis norvegica). Despite the non-selective feeding behaviour by T. longicornis and C. typicus, selectivity coefficients on D. acuta progressively decreased as food availability increased. Similar response was not observed for A. bifilosa, which displayed an even less selective behaviour. A. bifilosa had no significant negative effect on the net growth of D. norvegica at the lowest food concentration offered, whereas T. longicornis and C. typicus had significant negative effects on the net growth of D. acuta at low concentrations, similar to those observed in situ. Both species could potentially contribute as a substantial loss factor for Dinophysis spp. provided they are abundant at the onset of the blooms. The estimated grazing impact by the copepod populations was only considerable when C. typicus abundance was high and D. acuta population in sharp decline. Our results suggest that when high abundance of grazers and poor growth condition of prey populations prevail, grazing impact by copepods can contribute considerably to prevent Dinophysis spp. populations to grow or to cause the populations to decline. Okadaic acid was detected in the zooplankton size fraction at one occasion, but the concentration was far lower than the one expected from the ingested toxins. Thus, even if copepods may act as vectors of DSP-toxins to higher trophic levels, the amount of these toxins transported in the food web by copepods seems limited.     

The genus Dinophysis in the Southern Adriatic Sea

Abstract

Some preliminary results of a survey carried out along the Southern Adriatic coasts (Brindisi-S.M. Leuca) are reported in order to contribute to the knowledge of the phytoplankon community of this area. Ten Dinophysis species had been identified, four of them are known to be involved in DSP mussel contamination. Spatio-temporal distribution of Dinophysis spp. is shown.     

Guild structure in water mites (Unionicola spp.) inhabiting freshwater mussels: choice,competitive exclusion and sex

Summary Unionicolid water mites inhabit freshwater unionid mussels during the nymphal and adult stages of their life-cycle. Regular sampling of mussels from two sites in St. Mark's River, Fl. established that each of four species of water mite (Unionicola abnormipes, U. fossulata, U. serrata and U. formosa) occurred mainly in one or two of the mussel species available at each site.The role of preference for particular mussel species during host location was assessed for the first three mite species by choice experiments, in which mites were offered different mussel species simultaneously. In five out of six experiments, mites entered normally unused mussels as often as they did normally used ones. Additionally, a sexual difference in choice was found for U. fossulata, with males preferring one mussel species and females showing no preference. One mussel species, (Anodonta imbecilis), normally unused but chosen by mite species during the lab. experiments, is inhabited exclusively by the fourth mite species, U. formosa, in the field. An experiment showed that U. formosa excludes other mite species aggressively from Anodonta imbecilis.The results illustrate the sometimes misleading nature of simple sampling data as an indication of host specificity or host preference in parasites. They suggest also that the population dynamics of some parasites might be more fruitfully compared to unrelated, free-living species than to other parasites.     

Seasonal distribution of species of the toxic dinoflagellate genus Dinophysis in Maizuru Bay (Japan), with comments on their autofluorescence and attachment of picophytoplankton

The seasonal distribution of the dinoflagellate genus, Dinophysis, in Maizuru Bay, Japan, was investigated from May 1997 to December 1999. Seven species of Dinophysis were detected, including the toxic species of Dinophysis acuminata and D. fortii. The most dominant species wasD. acuminata, detected year-around and more abundantly during periods when water temperatures were between 15 and 18 °C. No relationship was found between cell abundance of Dinophysis spp. and concentrations of dissolved inorganic nutrients. Phycoerythrin containing nano- and picophytoplankton (cryptophytes and cyanobacteria), suspected to be prey of mixotrophic Dinophysis, were enumerated simultaneously. A clear relationship was not found among the cell abundances of Dinophysis spp. and nano- and picophytoplankton. Autofluorescence of Dinophysis spp. (mainly D. acuminata and D. fortii) under blue-light excitation was usually of a yellow-orange color. Occasionally, Dinophysis spp. had red autofluorescencing and yellow-orange autofluorescencing particles. The proportion of cells possessing red autofluorescence tended to be higher in the warm season. Numerous coccoid cells of picophytoplankton (ca. 1–2 μm in diameter) were found attached to the cell surface of D. acuminata, D. fortii, etc. and food vacuole-like structures also observed. These observations suggest there is a close relationship between mixotrophic Dinophysis spp. and certain picophytoplankton. Based on our observations, the possibility that the picophytoplankton found to be attached onto Dinophysis cell surfaces are a food source for Dinophysis, and a source of DSP toxins, is discussed.     

Seasonal variability of lipophilic toxins during a Dinophysis acuta bloom in Western Iberia: Differences between picked cells and plankton concentrates

This work describes and compares the seasonal variability of toxin profiles and content, estimated by LC–MS analyses, in picked cell of Dinophysis acuta Ehrenberg, in plankton concentrates rich in this species, and in extracellular lipophilic toxins collected by adsorbent resins during weekly sampling in a Galician ría (Western Iberia) from October 2005 to January 2006. Picked cells of D. acuta—which exhibited a fairly stable OA:DTX2 ratio, close to 3:2, but a variable okadaates:PTX2 ratio—showed a 9-fold variation in cell toxin quota, which was partly related to cellular volume, with maximum values (19 pg cell⁻¹) observed during the exponential decline of the population. Large differences in toxin profiles and content were observed between picked cells and plankton concentrates (up to 73 pg cell⁻¹ in the latter), that were most conspicuous after the bloom decline. The toxin profile of picked cells was more similar to that observed in the adsorbent resins than to the profiles of plankton concentrates. Their continued detection several weeks after the disappearance of Dinophysis spp. indicates that these toxins may take a long time to be degraded. It is concluded that analyses of picked-cells are essential to determine the contribution of each species of Dinophysis to a toxic outbreak. Estimates of cellular toxin content from plankton concentrates can lead to considerable overestimates after Dinophysis blooms decay due to extracellular toxins that persist in the water column, possibly bound to organic aggregates and detritus, and are retained (>0.22 μm) in the filters.     

Development of Molecular Probes for Dinophysis (Dinophyceae) Plastid: A Tool to Predict Blooming and Explore Plastid Origin

Dinophysis are species of dinoflagellates that cause diarrhetic shellfish poisoning. We have previously reported that they probably acquire plastids from cryptophytes in the environment, after which they bloom. Thus monitoring the intracellular plastid density in Dinophysis and the source cryptophytes occurring in the field should allow prediction of Dinophysis blooming. In this study the nucleotide sequences of the plastid-encoded small subunit ribosomal RNA gene and rbcL (encoding the large subunit of RuBisCO) from Dinophysis spp. were compared with those of cryptophytes, and genetic probes specific for the Dinophysis plastid were designed. Fluorescent in situ hybridization (FISH) showed that the probes bound specifically to Dinophysis plastids. Also, FISH on collected nanoplankton showed the presence of probe-hybridized eukaryotes, possibly cryptophytes with plastids identical to those of Dinophysis. These probes are useful not only as markers for plastid density and activity of Dinophysis, but also as tools for monitoring cryptophytes that may be sources of Dinophysis plastids.     

Population dynamics of potentially harmful algal blooms in Bizerte Lagoon,Tunisia

The population dynamics of potentially harmful phytoplankton in the semi-closed, coastal Bizerte Lagoon, Tunisia, in the south-western Mediterranean, were examined from November 2007 to February 2009 at six sampling stations, three situated in areas of mussel and oyster farming. The harmful species monitored included the potential producers of amnesic shellfish poisoning (Pseudo-nitzschia spp.), paralytic shellfish poisoning (Alexandrium spp.), diarrheic shellfish poisoning (Dinophysis spp. and Prorocentrum spp.), ichthyotoxins (Cochlodinium polykrikoides, Akashiwo sanguinea and Karenia mikimotoi), yessotoxins (Gonyaulax spinifera) and the discolouration of water (Neoceratium lineatum and Protoperidinium sp.). These were numerically dominated by potentially toxic species of the diatom genus Pseudo-nitzschia, which were present year-round at all stations. Prorocentrum spp., Dinophysis spp. and Neoceratium lineatum were the most abundant and recurrent harmful dinoflagellates, exhibiting their highest densities at the shellfish farming stations. Alexandrium spp. bloomed only on one occasion, reaching its highest densities at a shellfish farming station. Canonical correspondence analyses revealed significant relationships between the harmful phytoplankton species monitored and the environmental conditions. The widespread distribution of harmful phytoplankton in Bizerte Lagoon, with the permanent presence of Pseudo-nitzschia spp. and the high frequency of dinoflagellate blooms in the shellfish areas, suggests a potential risk of shellfish poisoning events in the region.     

SMALL CELL AND INTERMEDIATE CELL FORMATION IN SPECIES OF DINOPHYSIS (DINOPHYCEAE, DINOPHYSIALES)

Observations of two distinct size classes with similar shape in natural populations of Dinophysis Ehrenberg were first reported by Jorgensen in 1923 and intermediate forms exhibiting a continuum between the typical vegetative cell and a putative small cell by Wood in 1954. Focused attention on Dinophysis spp. associated with diarrhetic shellfish intoxications in the last decade has provided new examples of small cells in the genus, sometimes with contours dissimilar from the corresponding vegetative cells; dimorphic individuals; and large/small cell couplets. This work was based on in situ observations during intensive sampling for cell cycle studies of Dinophysis acuminata Claparéde et Lachmann, Dinophysis acuta Ehrenberg, Dinophysis caudata Saville-Kent, and Dinophysis tripos Gourret; on laboratory incubations of D. acuminata; and on a thorough search of documented information on morphological variability of Dinophysis spp. During in situ division, most dividing cells exhibit a normal longitudinal fission, but some (1%–10%) undergo a “depauperating” fission, leading to pairs of dimorphic cells with dissimilar moieties. After separation and sulcal list regeneration, these dimorphic cells become D. skagii Paulsen, D. dens Pavillard, D. diegensis Kofoid, and D. diegensis Kofoid var. curvata-like individuals, which can also be observed forming couplets D. acuminata/D. skagii, D. acuta/D. dens, and D. caudata/D. diegensis attached by their ventral margins. Small cells can grow again to large size, as shown in laboratory incubations of D. acuminata, thus partly explaining observations of thecal intercalary bands, and intermediate forms. The sexual nature of the small cells will not be unequivocally demonstrated until controlled germination of the alleged cyst forms is achieved, and some intermediate forms may correspond to undescribed stages after cyst germination. These observations suggest common patterns in the life cycle of Dinophysis spp. Intraspecific morphological variability of Dinophysis spp. in a given geographic area can largely be attributed to small cell formation, as a response to changing environmental conditions, and may be a part of the sexual cycle of these species. Small cells seem to be able to enlarge, leading to intermediate cell and further vegetative cell formation as part of a three-looped life history pattern in Dinophysis.     

Are the mitochondrial cox1 and cob genes suitable markers for species of Dinophysis Ehrenberg?

The identification of Dinophysis species with similar morphology but different toxic (Diarrhetic Shellfish Poisoning, DSP) potential is a crucial task in harmful algae monitoring programmes. The taxonomic assignment of Dinophysis species using molecular markers is a difficult task due to extremely low interspecific variability within their nuclear ribosomal genes and intergenic regions. Mitochondrial cox1 gene has been proposed as a better specific marker for Dinophysis species based on its higher resolution for two morphologically related species (Dinophysis acuminata and Dinophysis ovum) of the “Dinophysis acuminata complex”. In this study, the potential of two mitochondrial genes (mt cox1 and cob) to discriminate among six Dinophysis species (field isolates and cultures) associated with DSP events was explored. Neither mt cox1 nor cob genes provided enough resolution for all species of Dinophysis. The cob gene showed very poor resolution and grouped all Dinophysis spp. in a common clade. In contrast, the cox1 phylogeny distinguished 5 clades in the Dinophysiales – the “acuminata complex”, the “caudata group”, “acuta + norvegica” and Phalacromaspp. However, within the “D. acuminata complex” mtcox1 is so far the unique marker that differentiates D. acuminata from other species: isolates of D. ovum and Dinophysis sacculus had almost identical sequences (only four mismatches), but they were well separated from D. acuminata. D. acuminata and Dinophysis skagii (considered a life cycle stage of the former) showed identical cox1 sequences. Probes towards this gene can be useful in Mediterranean and Western Iberia sites where the co-occurrence of close morphotypes of D. acuminata and D. sacculus pose a problem for monitoring analyses. This is the first report on cultures of D. sacculus and its phylogenetic relation with other species of the D. acuminata complex.     

High performance liquid chromatography analysis of diarrhetic toxins inDinophysis spp. from the French coast

Diarrhetic shellfish poisoning has been a recurrent problem along the Brittany coast (France) since 1983. Okadaic acid (O.A.), the main toxin detected in mussels, is generally associated with the presence ofDinophysis cells in sea water. We report here the results of okadaic acid analyses by high performance liquid chromatography, on planktonic samples collected during the summer of 1991.D. sacculus, the majorDinophysis species, throughout this period, showed low okadaic acid content in raw extract, whereas toxic levels were 3- to 12-fold higher in sorted samples than in raw ones. A maximum O.A. concentration of 29.6 pg cell^-1 was found inD. sacculus/D. cf. acuminata extract. Similarly, higher O.A. levels were noted in raw samples when these two species were associated. Generally speaking, variations in raw samples were similar to those in sorted samples. Nevertheless, whenD. rotundata reached a concentration equal to that ofD. sacculus in sorted samples, the O.A. level was lower.     

Real-time PCR Detection of Dinophysis Species in Irish Coastal Waters

Diarrhetic shellfish toxin-producing Dinophysis species occur in Irish coastal waters throughout the year. Dinophysis acuta and Dinophysis acuminata are the most commonly occurring species and are responsible for the majority of closures of Irish mussel farms. This study describes the development of a qualitative real-time polymerase chain reaction (PCR) assay for identification of D. acuta and D. acuminata in Irish coastal waters. DNA sequence information for the D1-D2 region of the large ribosomal sub-unit (LSU) was obtained, following single-cell PCR of D. acuta and D. acuminata cells isolated from Irish coastal locations. PCR primers and hybridization probes, specific for the detection of D. acuta, were designed for real-time PCR on the LightCycler™. The LightCycler™ software melt curve analysis programme determined that D. acuta was identified by a melt-peak at 61°C, while D. acuminata cells produced a melt peak at 48°C. The limit of detection of the real-time PCR assay was determined to be one to ten plasmid copies of the LSU D1-D2 target region for both species and one to five D. acuminata cells. Lugol's preserved water samples were also tested with the assay. The real-time PCR assay identified Dinophysis species in 100% of samples found to contain Dinophysis species by light microscopy and had a greater than 90% correlation with light microscopy for identification of D. acuta and D. acuminata in the samples. The assay can identify and discriminate D. acuta and D. acuminata at low numbers in Irish waters and has the potential to add value to the Irish phytoplankton monitoring programme.     

GENETIC VARIABILITY AND MOLECULAR PHYLOGENY OF DINOPHYSIS SPECIES (DINOPHYCEAE) FROM NORWEGIAN WATERS INFERRED FROM SINGLE CELL ANALYSES OF rDNA1

The objectives of this study were to determine rDNA sequences of the most common Dinophysis species in Scandinavian waters and to resolve their phylogenetic relationships within the genus and to other dinoflagellates. A third aim was to examine the intraspecific variation in D. acuminata and D. norvegica, because these two species are highly variable in both morphology and toxicity. We obtained nucleotide sequences of coding (small subunit [SSU], partial large subunit [LSU], 5.8S) and noncoding (internal transcribed spacer [ITS]1, ITS2) parts of the rRNA operon by PCR amplification of one or two Dinophysis cells isolated from natural water samples. The three photosynthetic species D. acuminata, D. acuta, and D. norvegica differed in only 5 to 8 of 1802 base pairs (bp) within the SSU rRNA gene. The nonphotosynthetic D. rotundata (synonym Phalacroma rotundatum[Claparède et Lachmann] Kofoid et Michener), however, differed in approximately 55 bp compared with the three photosynthetic species. In the D1 and D2 domains of LSU rDNA, the phototrophic species differed among themselves by 3 to 12 of 733 bp, whereas they differed from D. rotundata by more than 100 bp. This supports the distinction between Dinophysis and Phalacroma. In the phylogenetic analyses based on SSU rDNA, all Dinophysis species were grouped into a common clade in which D. rotundata diverged first. The results indicate an early divergence of Dinophysis within the Dinophyta. The LSU phylogenetic analyses, including 4 new and 11 Dinophysis sequences from EMBL, identified two major clades within the phototrophic species. Little or no intraspecific genetic variation was found in the ITS1–ITS2 region of single cells of D. norvegica and D. acuminata from Norway, but the delineation between these two species was not always clear.     

Epiphytic abundance and toxicity of Prorocentrum lima populations in the Fleet Lagoon, UK

Planktonic Dinophysis spp. and epiphytic Prorocentrum lima (Ehrenberg) Dodge are known dinoflagellate producers of okadaic acid (OA) and dinophysistoxins (DTX), causative phycotoxins of diarrhetic shellfish poisoning (DSP). Underestimation of toxic dinoflagellates associated with a toxic event may be due to the lack of sampling of species with epiphytic and epibenthic strategies, such as P. lima. As Dinophysis spp. is not found in the Fleet Lagoon, Dorset, but previous DSP events have closed the Crassostrea gigas oyster farm, P. lima is the most likely causative organism. A field assay for separating microalgal epiphytes and concentrating wild cells on to filters was successfully applied to sub-samples of a variety of macroalgae and macrophytes (seagrass) collected from the Fleet during summer 2002. P. lima was present in increasing cell densities on most substratum species, over the sampling period, from 10² to 10³ cells g⁻¹ fresh weight (FW) plant biomass. LC–MS analysis detected OA and DTX-1 in extracts of wild P. lima cells, in ratios characteristic of P. lima strains previously isolated from the Fleet. No toxins, however, were detected in oyster flesh.     

Contribution to toxicity assessment of <Emphasis Type="BoldItalic">Dinophysis acuminata</Emphasis> (Dinophyceae)

Blooms of Dinophysis in French coastal waters are implicated in most bans on marketing commercial bivalves. However, the relation between Dinophysis cell density and shellfish toxicity is not always consistent. Discrepancies may be due to the simple fact that it is nearly impossible to compare an integral over a few days (shellfish toxin content) and water samples. Furthermore, it seems that cells may have a variable specific toxicity. This work focuses on the variability in cell toxicity taking into account recent findings and using liquid chromatography coupled to mass spectrometry with an ion trap and electrospray interface. Esterified analogues of okadaic acid (DTX-4 and diol-esters) have been identified in cultures of Prorocentrum lima, another okadaic acid producer. These analogues are inactive on some protein phosphatases, contrary to okadaic acid, and seem to protect the cell from harmful effects by the toxin and to be enzymatically hydrolyzed during cell lysis. In order to document specific toxicity and to validate the presence of these analogues, D. acuminata concentrates were subjected to two separate heating and freeze/thaw procedures, respectively inhibiting or promoting hydrolysis. This paper reports on the high variability of D. acuminata specific toxicity and the presence of esters found in half of the samples only.     

Relationships between occurrences of toxic Dinophysis species (Dinophyceae) and small phytoplanktons in Japanese coastal waters

Fluctuations of the genus Dinophysis, which contained several toxic species of diarrhetic shellfish poisoning (DSP), were investigated during blooms in Hiroshima Bay, Mutsu Bay and Ise Bay, Japan. The co-occurrences of small phytoplanktons (cryptophytes, other nanophytoplanktons, cyanobacteria and eukaryotic picophytoplanktons) were investigated to search for relationships with mixotrophic Dinophysis. Cryptophytes were divided into three size-groups based on length of their chloroplasts (>10, 5–10 and <5 μm) during counting by epifluorescence microscopy. Clear relationships were not found between the occurrences of Dinophysis spp. and nanophytoplanktons, cyanobacteria and eukaryotic picophytoplanktons. However, the fluctuations of small-sized cryptophytes (<5 μm) showed a close relationship with that of D. acuminata in Hiroshima Bay. In Mutsu Bay, small-sized cryptophytes also accompanied the first occurrence peak of Dinophysis spp. In Ise Bay, peaks of the occurrences of middle- and small-sized cryptophytes were observed 2–3 weeks before the peak of D. acuminata. These cryptophytes decreased rapidly with increase in D. acuminata. These results suggest the possibility that small-sized cryptophytes may be food organisms for mixotrophic Dinophysis, with the abundance of Dinophysis dependent on these cryptophytes.