Maria A. Faust

Observation of sand-dwelling toxic dinoflagellates (Dinophyceae) from widely differing sites, including two new species

Dinoflagellate associations, including toxic and potentially toxic benthic species, were examined in sand from South Water Cay and Carrie Bow Cay, Belize. The inshore sand habitat in localized areas of warm shallow lagoonal waters supported blooms of toxic assemblages of dinoflagellates. In the sand, the dominant microalgae were dinoflagellates; cyanobacteria were a minor component and diatoms were absent. Ciliates and nematodes were present. Assemblages of microorganisms in colored sand were examined for 4 consecutive days after which a storm washed away the patch. The sand‐dwelling dinoflagellate assemblage included 16 species where densities ranged from as low as 1.3% to 15% of total cell densities. The dominant species was Scrippsiella subsalsa, having 1.8 × 105 to 2.6 × 105 cells g‐1 sand. Toxic dinoflagellates identified in the sand were Gambierdiscus toxicus, Ostreopsis lenticularis, Prorocentrum lima, Prorocentrum mexicanum, and Amphidinium carteri. The potentially toxic Ostreopsis labens, Gambierdiscus belizeanussp. nov., and Coolia tropicalis sp. nov. were also identified. Toxic and potentially toxic species represented 36% to 60% of total microalgal cell assemblage.

Taxonomy of Gambierdiscus including four new species, Gambierdiscus caribaeus, Gambierdiscus carolinianus, Gambierdiscus carpenteri and Gambierdiscus ruetzleri (Gonyaulacales, Dinophyceae)

Litaker R.W., Vandersea M.W., Faust M.A., Kibler S.R., Chinain M., Holmes M.J., Holland W.C. and Tester P.A. 2009. Taxonomy of Gambierdiscus including four new species, Gambierdiscus caribaeus, Gambierdiscus carolinianus, Gambierdiscus carpenteri and Gambierdiscus ruetzleri (Gonyaulacales, Dinophyceae). Phycologia 48: 344–390. DOI: 10.2216/07-15.1 Gambierdiscus species produce toxins that cause ciguatera fish poisoning (CFP), the most common nonbacterial illness associated with fish consumption worldwide. Understanding the role that individual Gambierdiscus species play in causing CFP is hampered because the morphologically similar species in the genus are difficult to distinguish. Ambiguities in the description of the type species Gambierdiscus toxicus also exist. This paper presents detailed line drawings along with additional scanning electron microscopy (SEM) and light micrographs to more fully characterize the six known Gambierdiscus species in addition to describing four new species, Gambierdiscus caribaeus sp. nov., Gambierdiscus carolinianus sp. nov., Gambierdiscus carpenteri sp. nov., and Gambierdiscus ruetzleri sp. nov. All four of the new species are photosynthetic and epibenthic and have a Kofoidian plate formula of Po, 3′, 7″, 6C, 6 or 7S, 5′″, 1p, and 2″″. Establishment of these species is supported by both morphological differences and three independent phylogenetic analyses utilizing small-subunit, as well as D1–D3 and D8–D10 large-subunit rDNA gene sequences. During the course of this study it became apparent that the description of the type species, G. toxicus, included more than one species. Since no type material exists we propose that Fig. 1 in Adachi and Fukuyo (1979) be designated as the lectotype for this species and that the epithecal view of isolate GTT-91 shown in Fig. 1 from Chinain et al. (1999) be designated as the epitype. The GTT-91 isolate has been well characterized genetically and the original SEM stub of this isolate archived at the Smithsonian Institution has been re-examined to provide a more detailed morphological analysis of the epitype.

FURTHER SEM STUDY OF MARINE DINOFLAGELLATES : THE GENUS OSTREOPSIS (DINOPHYCEAE)

This paper presents a comprehensive examination of the taxonomy of the genus Ostreopsis Schmidt. The morphology of six species of marine dinoflagellates, Ostreopsis siamensis Schmidt 1902. Ostreopsis lenticularis Fukuyo 1981, Ostreopsis ovata Fukuyo 1981, Ostreopsis heptagona Norris, Bomber, et Balech 1985, Ostreopsis mascarenensis Quod 1994, and Ostreopsis labens Faust et Morton 1995 from three geographical regions (Japan, Southwest Indian Ocean, and the Caribbean) and three marine habitats (sand, water column, and macroalgal surfaces) are described from scanning electron micrographs. Differences in the following morphological characteristics differentiated the species: cell shape and size, and ornamentation of the epitheca, cingulum, and hypotheca. The thecal plate formula of the six Ostreopsis species is Po, 3′, 7″, 6C, 6S?, Vp, Rp, 5′″, 1p, 2″″, with differences in thecal plate size and shape. The cingulum in ventral view has two prominent structures: a ventral plate (Vp) with a ventral pore (Vo) and a ridged plate (Rp) that distinguishes Ostreopsis species from any other dinoflagellate taxa. This paper also includes ecological and toxicity information regarding the six Ostreopsis species.

Prorocentrum belizeanum, Prorocentrum elegans, and Prorocentrum caribbaeum, three new benthic species (Dinophyceae) from a mangrove island, Twin Cays, Belize

Three new benthic dinoflagellate species, Prorocentrum belizeanum, Prorocentrum elegans, and Prorocentrum caribbaeum, from mangrove floating detritus are described from scanning electron micrographs. Species were identified based on shape, size, surface micromorphology, ornamentation of thecal plates, and architecture of the periflagellar area and intercalary band.

THREE NEW BENTHIC SPECIES OF PROROCENTRUM (DINOPHYCEAE) FROM CARRIE BOW CAY, BELIZE: P. SABULOSUM SP. NOV., P. SCULPTILE SP. NOV., AND P. ARENARIUM SP. NOV.1

Three new benthic, sand‐dwelling dinqflagellate species, Prorocentrum sabulosum, Prorocentrum scuptile, and Prorocentrum arenarium, from coral rubble are described from scanning electron micrographs. Species were identified based on shape, size, surface micromorphology, ornamentation of thecal plates, and architecture of the periflagellar area and intercalary band.

MORPHOLOGY AND ECOLOGY OF THE MARINE DINOFLAGELLATE OSTREOPSIS LABENS SP. NOV. (DINOPHYCEAE)1

A new species, Ostreopsis labens Faust et Morton sp. nov., is described from three marine habitats: lagoonal water and lagoonal sand from the barrier reef of Belize, and associated with macroalgae from coral reef habitats of Oshigaki and Iriomote Islands, Japan. Dimensions of Ostreopsis labens cells are 60–86 μm long, 70–80 μm wide, and 81–110 μm in dorsoventral depth. Cells are broadly ovoid, anterioposteriorly compressed bearing a spherical nucleus and many chloroplasts. The epitheca is convex and composed of three apical plates, seven precingular plates, and an apical pore plate. The cingulum is composed of six plates. The hypotheca is constructed of five postcingular plates, one posterior intercalary, and two antapical plates. The sulcus is small, recessed, and hidden and exhibits a ventral pore and a ridged, curved plate. The thecal arrangement of O. labens is Po, 3′, 7″ 6C, 6S(?), Vp, Rp, 5″, 1p, 2″. Only one sulcal list is present. The thecal plates have a smooth surface with distinct round pores. The intercalary band between the thecal plates is smooth. A row of marginal pores line the lipped cingulum. Ostreopsis species are anteroposteriorly flattened, photosynthetic, benthic dinoflagellates that are more diverse in ecology than previously known. Ostreopsis labens is capable of living in three marine habitats: in the water column, in sand, and on macroalgal surfaces. It was most numerous in sand and less in lagoonal waters, and only a few cells were associated with macroalgae. Light and scanning electron microscopy studies revealed engulfed cells within O. labens, which indicates mixotrophic/phagotrophic behavior. A ventral opening situated in the cingulum of O. labens exhibits size variability; it may serve as an opening for engulfiing food particles because it varies in size. We propose that ingestion of prey by O. labens occurs through the ventral opening, the proposed feeding apparatus of this species, which is similar to the function of the peduncle‐like structure of mixotrophic dinoflagellates. The behavior of O. labens appears similar to that previously described for Dinophysis species.

DEVELOPMENT OF SEMI-QUANTITATIVE PCR ASSAYS FOR THE DETECTION AND ENUMERATION OF GAMBIERDISCUS SPECIES (GONYAULACALES, DINOPHYCEAE)(1).

Ciguatera fish poisoning (CFP) is a serious health problem in tropical regions and is caused by the bioaccumulation of lipophilic toxins produced by dinoflagellates in the genus Gambierdiscus. Gambierdiscus species are morphologically similar and are difficult to distinguish from one another even when using scanning electron microscopy. Improved identification and detection methods that are sensitive and rapid are needed to identify toxic species and investigate potential distribution and abundance patterns in relation to incidences of CFP. This study presents the first species‐specific, semi‐quantitative polymerase chain reaction (qPCR) assays that can be used to address these questions. These assays are specific for five Gambierdiscus species and one undescribed ribotype. The assays utilized a SYBR green format and targeted unique sequences found within the SSU, ITS, and the D1/D3 LSU ribosomal domains. Standard curves were constructed using known concentrations of cultured cells and 10‐fold serial dilutions of rDNA PCR amplicons containing the target sequence for each specific assay. Assay sensitivity and accuracy were tested using DNA extracts purified from known concentrations of multiple Gambierdiscus species. The qPCR assays were used to assess Gambierdiscus species diversity and abundance in samples collected from nearshore areas adjacent to Ft. Pierce and Jupiter, Florida USA. The results indicated that the practical limit of detection for each assay was 10 cells per sample. Most interestingly, the qPCR analysis revealed that as many as four species of Gambierdiscus were present in a single macrophyte sample.

PROROCENTRUM LEVIS, A NEW BENTHIC SPECIES (DINOPHYCEAE) FROM A MANGROVE ISLAND, TWIN CAYS, BELIZE

As part of a long‐term study of benthic dinoflagellates from the Belizean barrier reef system, we report a new species: Prorocentrum levis M. A. Faust, Kibler, Vandersea, P. A. Tester et Litaker sp. nov. P. levis cells are oval in valve view and range in size from 40 to 44 μm long and 37 to 40 μm wide. Each valve surface is smooth, with 221–238 valve pores and 99–130 marginal pores. These pores are uniformly small and range in diameter from 0.13 to 0.19 μm. Asexual reproduction in P. levis is atypical, occurring within a hyaline envelope, and produces long branching chains of adherent cells. A phylogenetic analysis of SSU rDNA indicated that of the Prorocentrum species sequenced so far, P. levis was most closely related to P. concavum. P. levis produces okadaic acid and dinophysis toxin‐2 (DTX2). Further, SEM observations and SSU rDNA sequence for P. belizeanum M. A. Faust, which was isolated at the same time, are also presented.

Associations of microalgae and meiofauna in floating detritus at a mangrove island, Twin Cays, Belize

Abstract Associations of benthic microalgae and meiofauna affected by temperature, salinity and dissolved oxygen concentrations were examined in floating detritus in a shallow mangrove embayment in a 6 day time-series investigation. Floating detritus exhibits a diurnal movement: it rises to the surface via oxygen bubbles generated by attached microalgae at sunrise and sinks down at sunset. In floating mangrove detritus, dinoflagellates were present in highest proportion (50–90%), followed by diatoms (5–15%), cyanobacteria (3–25%) and dinoflagellate cysts (1–7%). Microalgal densities correlated significantly with dissolved oxygen concentrations (r2 = 0.763, P

A FURTHER SEM STUDY OF MARINE BENTHIC DINOFLAGELLATES FROM A MANGROVE ISLAND, TWIN CAYS, BELIZE, INCLUDING PLAGIODINIUM BELIZEANUM GEN. ET SP. NOV.1

This study indicates that bilaterally flattened, armored, benthic dinoflagellates are more diverse in morphology than previously known. A new species, Plagiodinium belizeanum Faust et Balech gen. et. sp. nov., is described in floating detritus from Twin Cays, Belize, mangrove habitats. Plagiodinium belizeanum cells are small, with dimensions of 26.5–30.5 μm in length, 20–24.5 μm in width, and 6.5–8.5 μm in depth. Cells are oblong and bilaterally compressed with a posteriorly located, spherical nucleus, many chloroplasts, and spherical starch granules. The epitheca descends ventrally, is cap‐shaped, and is composed of five plates and a very small platelet provisionally named P0 situated in the center. The epitheca is narrowly oval in apical view with a pointed truncated ventral side and a rounded dorsal side. The cingulum is composed of five plates. The hypotheca is constructed of five posteriorly elongated postcingular plates and one antapical plate. The sulcus is very short and narrow, comprised of five very small plates. The thecal plate arrangement of P. belizeanum is P0, 5′, O″, 5C, 5″′, 1″″, 5S. No lists are present. Thecal plates have a smooth surface with small and irregularly scattered pores. The intercalary band is smooth on outer cell surface and broadly striated on its inner surface. We conclude that P. belizeanum represents a new, benthic, peridinioid, armored genus, Plagiodinium gen. nov. The taxonomic position of P. belizeanum sp. nov. is compared to related sand‐dwelling and bilaterally flattened benthic dinoflagellates.