Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Growth, Toxin Content and Production of Dinophysis norvegica in Cultured Strains Isolated From Japan

Version 1 : Received: 28 February 2023 / Approved: 3 March 2023 / Online: 3 March 2023 (09:33:50 CET)

A peer-reviewed article of this Preprint also exists.

Nagai, S.; Basti, L.; Uchida, H.; Kuribayashi, T.; Natsuike, M.; Sildever, S.; Nakayama, N.; Lum, W.M.; Matsushima, R. Growth, Toxin Content and Production of Dinophysis Norvegica in Cultured Strains Isolated from Funka Bay (Japan). Toxins 2023, 15, 318. Nagai, S.; Basti, L.; Uchida, H.; Kuribayashi, T.; Natsuike, M.; Sildever, S.; Nakayama, N.; Lum, W.M.; Matsushima, R. Growth, Toxin Content and Production of Dinophysis Norvegica in Cultured Strains Isolated from Funka Bay (Japan). Toxins 2023, 15, 318.

Abstract

This study is the first report of the successful cultivation of Dinophysis norvegica isolated from Japanese coastal waters (7 of 48 isolated cells were established as the clonal cultures, 14.5%) and their toxin content and production. The strains were maintained at relatively high abundance (>2,000 cells mL-1) for more than one year when fed on Mesodinium rubrum with the addition of Teleaulax amphioxeia. At the end of the one-month incubation, the total amounts of pectenotoxin-2 (PTX2), dinophysistoxin-1 (DTX1) and okadaic acid (OA) were 132.0-375.0 ng mL-1 (n = 7), 0.7-3.6 ng mL-1 (n = 3) and trace level OA (n = 1), respectively. Similarly, the cell quota of PTX2 and DTX1 were 60.6-152.4 pg cell-1 (n = 7) and 0.5-1.2 pg cell-1 (n = 3), respectively. These data indicated that toxin production varies depending on the strains in this species. In the growth experiment, D. norvegica grew, but it was very slow first 12 days. However, after that they grew exponentially with the maximum growth rate of 0.56 divisions day-1 (during Day 24-27), reaching a maximum concentration of 3,000 cells mL-1 at the end of incubation (Day 36), suggesting that they have a long lag phase. In the toxin production, OA was not at a detectable level (≤ 0.010 ng mL-1) during the 36 days of incubation except for Day 6. The concentration of DTX1 and PTX2 showed similar patterns as described for vegetative growth, but the toxin production still increased on Day 36 (1.3 ng mL-1 and 154.7 ng mL-1 in DTX1 and PTX2, respectively). The findings of this study provide novel information on the toxin content and production in D. norvegica as well as details on the culturing and maintenance of the species.

Keywords

Diarrhetic shellfish poisoning (DSP); Dinophysis norvegica; Mesodinium rubrum; Teleaulax amphioxeia; Pectenotoxin-2 (PTX2); Dinophysistoxin-1 (DTX1); Okadaic acid (OA)

Subject

Environmental and Earth Sciences, Environmental Science

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