Search

Among dinoflagellates responsible for benthic harmful algal blooms, the genus Ostreopsis primarily described from tropical areas has been increasingly reported from subtropical and temperate areas worldwide. Several species of this toxigenic genus produce analogs of palytoxin, thus representing a major threat to human and environmental health. The taxonomy of several species needs to be clarified as it was based mostly on morphological descriptions leading in some cases to ambiguous interpretations and misidentifications. The present study aims at reporting a benthic bloom that occurred in April 2019 in Tahiti island, French Polynesia. A complete taxonomic investigation of the blooming Ostreopsis species was realized using light, epifluorescence and field emission electron microscopy and phylogenetic analyses inferred from LSU rDNA and ITS–5.8S rDNA regions. Toxicity of a natural sample and strains isolated from the bloom was assessed using both neuroblastoma cell-based assay and LC-MS/MS analyses. Morphological observations showed that cells were round to oval, large, 58.0–82.5 µm deep (dorso-ventral length) and 45.7–61.2 µm wide. The cingulum was conspicuously undulated, forming a ‘V’ in ventral view. Thecal plates possessed large pores in depressions, with a collar rim. Detailed observation also revealed the presence of small thecal pores invisible in LM. Phylogenetic analyses were congruent and all sequences clustered within the genotype Ostreopsis sp. 6, in a subclade closely related to sequences from the Gulf of Thailand and Malaysia. No toxicity was found on the field sample but all the strains isolated from the bloom were found to be cytotoxic and produced ostreocin D, a lower amount of ostreocins A and B and putatively other compounds. Phylogenetic data demonstrate the presence of this species in the Gulf of Thailand, at the type locality of O. siamensis, and morphological data are congruent with the original description and support this identification.

To date, the genus Ostreopsis comprises eleven described species, of which seven are toxigenic and produce various compounds presenting a major threat to human and environmental health. The taxonomy of several of these species however remains controversial, as it was based mostly on morphological descriptions leading, in some cases, to ambiguous interpretations and even possible misidentifications. The species Ostreopsis lenticularis was first described by Y. Fukuyo from French Polynesia using light microscopy observations, but without genetic information associated. The present study aims at revisiting the morphology, molecular phylogeny and toxicity of O. lenticularis based on the analysis of 47 strains isolated from 4 distinct locales of French Polynesia, namely the Society, Australes, Marquesas and Gambier archipelagos. Observations in light, epifluorescence and field emission scanning electron microscopy of several of these strains analyzed revealed morphological features in perfect agreement with the original description of O. lenticularis. Cells were oval, not undulated, 60.5–94.4 μm in dorso-ventral length, 56.1–78.2 μm in width, and possessed a typical plate pattern with thecal plates showing two sizes of pores. Phylogenetic analyses inferred from the LSU rDNA and ITS–5.8S sequences revealed that the 47 strains correspond to a single genotype, clustering with a strong support with sequences previously ascribed to Ostreopsis sp. 5. Clonal cultures of O. lenticularis were also established and further tested for their toxicity using the neuroblastoma cell-based assay and LC MS/MS analyses. None of the 19 strains tested showed toxic activity on neuroblastoma cells, while LC MS/MS analyses performed on the strains from Tahiti Island (i.e. type locality) confirmed that palytoxin and related structural analogs were below the detection limit. These findings allow to clarify unambiguously the genetic identity of O. lenticularis while confirming previous results from the Western Pacific which indicate that this species shows no toxicity, thus stressing the need to reconsider its current classification within the group of toxic species.

Chattonella species in a Mediterranean lagoon (Santa Giusta Lagoon, Sardinia, Italy) were identified by applying a molecular approach to fixed natural phytoplankton samples collected over the last two decades. Like the other raphidophytes, Chattonella cells are naked and lose their shape when fixed, making species identification difficult on the basis of their morphological characteristics. Employing species-specific primers (oBTG-005-F, oBTG-027-R, oBTG-028-R) for the amplification of the ITS-5.8S rDNA region, we established the occurrence of C. subsalsa in fixed natural phytoplankton samples collected in coincidence with fish death events. Additionally, we established the presence of the recently discovered C. cf. subsalsa Adriatic genotype by analysing cellular cultures obtained from the same lagoon in 2013. This is the second worldwide record of C. cf. subsalsa Adriatic genotype. Our results revealed that the species-specific primers oBTG-005-F and oBTG-028-R distinguished this new genotype only when present singularly. This study provides valuable data that increase knowledge of C. subsalsa genotypes and of the long-term occurrence of Chattonella blooms in a transitional ecosystem through the use of samples up to 20 years old., Se identificaron especies de Chattonella mediante la aplicación de técnicas moleculares en muestras naturales de fitoplancton. Las muestras fueron recogidas y fijadas durante las últimas dos décadas en una laguna litoral mediterránea (Laguna de Santa Giusta, Cerdeña, Italia). Al igual que otras rafidoficeas, las células de Chattonella no poseen teca y, por lo tanto, pierden su forma cuando se fijan lo que dificulta la identificación basada en características morfológicas. Con el uso de cebadores específicos a nivel de especie (OBTG-005-F, OBTG-027-R, OBTG-028-R) diseñados para la amplificación de la región ITS- 5.8S rDNA, se detectó la presencia de C. subsalsa en las muestras recogidas en periodos coincidentes con eventos de muerte de peces. A través del análisis de los cultivos celulares obtenidos de la misma laguna en el año 2013, se identificó la presencia, por segunda vez a nivel mundial, del recientemente descubierto genotipo Adriático de C. cf. subsalsa. Los resultados revelaron que los cebadores oBTG-005-F y oBTG-028-R amplifican este nuevo genotipo sólo cuando está presente individualmente. En este estudio se presentan datos relevantes para el conocimiento de los genotipos de C. subsalsa y sobre la presencia recurrente de proliferaciones de especies de Chatonella en un ecosistema de transición a través de la utilización de muestras recogidas durante los últimos veinte años y analizadas hoy en día.

Chattonella species in a Mediterranean lagoon (Santa Giusta Lagoon, Sardinia, Italy) were identified by applying a molecular approach to fixed natural phytoplankton samples collected over the last two decades. Like the other raphidophytes, Chattonella cells are naked and lose their shape when fixed, making species identification difficult on the basis of their morphological characteristics. Employing species-specific primers (oBTG-005-F, oBTG-027-R, oBTG-028-R) for the amplification of the ITS-5.8S rDNA region, we established the occurrence of C. subsalsa in fixed natural phytoplankton samples collected in coincidence with fish death events. Additionally, we established the presence of the recently discovered C. cf. subsalsa Adriatic genotype by analysing cellular cultures obtained from the same lagoon in 2013. This is the second worldwide record of C. cf. subsalsa Adriatic genotype. Our results revealed that the species-specific primers oBTG-005-F and oBTG-028-R distinguished this new genotype only when present singularly. This study provides valuable data that increase knowledge of C. subsalsa genotypes and of the long-term occurrence of Chattonella blooms in a transitional ecosystem through the use of samples up to 20 years old. Se identificaron especies de Chattonella mediante la aplicación de técnicas moleculares en muestras naturales de fitoplancton. Las muestras fueron recogidas y fijadas durante las últimas dos décadas en una laguna litoral mediterránea (Laguna de Santa Giusta, Cerdeña, Italia). Al igual que otras rafidoficeas, las células de Chattonella no poseen teca y, por lo tanto, pierden su forma cuando se fijan lo que dificulta la identificación basada en características morfológicas. Con el uso de cebadores específicos a nivel de especie (OBTG-005-F, OBTG-027-R, OBTG-028-R) diseñados para la amplificación de la región ITS- 5.8S rDNA, se detectó la presencia de C. subsalsa en las muestras recogidas en periodos coincidentes con eventos de muerte de peces. A través del análisis de los cultivos celulares obtenidos de la misma laguna en el año 2013, se identificó la presencia, por segunda vez a nivel mundial, del recientemente descubierto genotipo Adriático de C. cf. subsalsa. Los resultados revelaron que los cebadores oBTG-005-F y oBTG-028-R amplifican este nuevo genotipo sólo cuando está presente individualmente. En este estudio se presentan datos relevantes para el conocimiento de los genotipos de C. subsalsa y sobre la presencia recurrente de proliferaciones de especies de Chatonella en un ecosistema de transición a través de la utilización de muestras recogidas durante los últimos veinte años y analizadas hoy en día.