Akira Ishikawa, Yumi Takeichi. Setsuko Sakamoto, Mineo Yamaguchi
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1 Akira Ishikawa, Yumi Takeichi (Mie University) Setsuko Sakamoto, Mineo Yamaguchi (National Researchi Institute of Fisheries and Environment of Inland Sea)
2 Background Gambierdiscus spp. is the benthic and epiphytic dinoflagellate, commonly found on the macroalgae in coral reefs. Some of the Gambierdiscus spp. have been confirmed to produce maitotoxins and ciguatoxins, being the primary causative organism of ciguatera fish poisoning (CFP).
3 Background They are distributed widely in tropical and subtropical regions, where CFP mainly occurs. It has been reported that > 50,000 people suffer from CFP annually.
4 45 N 40 In Japan CFP is endemic in the subtropical regions, Okinawa & Amami Islands. Several CFP incidents have sporadically occurred in temperate regions. Russia Khabarovsk Japan 35 Temperate regions 30 Pacific Ocean 25 Okinawa/Amami Is. Subtropical regions E
5 45 N In Japan CFP is endemic in the subtropical region, Okinawa &Amami Islands. 40 Several CFP incidents have sporadically occurred in temperate region. Pacific Ocean 35 Temperate regions In Kii peninsula, CFP occurred in 2007, for the first time, and subsequently in Okinawa/Amami Is. Subtropical regions E
6 Sites ( ) show where the presence of the genus Gambierdiscus has been reported in temperate regions.
7 Having recent CFP incidents in Kii Peninsula, We examined for the presence of the genus Gambierdiscus in Ago Bay. Osaka Mie Nagoya Kii Peninsula Ago Bay Google map
8 Gambierdiscus found in Ago Bay Oblique ventral view A B 2 3 Po Epitheca Hypo heca C 4 1p D Apical pore Plate (Po) 3 2 Scal bars: 10 Main plate formula Po, 3, 7, 5, 1p, 2 (Ishikawa & Kurashima 2010: Bull Jpn Soc Fish Oceanogr)
9 Flowchart to distinguish the various Gambierdiscus species A 7 1 B C 4 1p Fig. 10 in et al. (2009): Phycologia 3 2 Scal bars: 10
10 99 51 Result of phylogenetic analysis (NJ) AY Prorocentrum micans LSU X Prorocentrum micans LSU AY : Akashiwo sangui... AY : Cochlodinium po AF : Gonyaulax polye... AY : Alexandrium min... AY : Alexandrium aff... AY : Alexandrium cat DQ : Alexandrium fun... DQ : Alexandrium tam Utilizing the regions D8 D10 of the LSU rdna gene EU Gambierdiscus pacificus c... EU Gambierdiscus pacificus c... EU Gambierdiscus pacificus c... EU Gambierdiscus pacificus c... EU Gambierdiscus pacificus c... EU Gambierdiscus pacificus c... EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo EU Gambierdiscus 84 belizeanus... EU Gambierdiscus belizeanus... EU Gambierdiscus belizeanus... EU Gambierdiscus belizeanus... EU Gambierdiscus belizeanus... EU Gambierdiscus belizeanus... EU Gambierdiscus belizeanus... EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo EU Gambierdiscus sp. carpent... EU Gambierdiscus sp. carpent... EU Gambierdiscus sp. carpent... EU Gambierdiscus sp. carpent... EU Gambierdiscus sp. carpent... EU Gambierdiscus sp. carpent... EU Gambierdiscus sp. carpent... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... EU Gambierdiscus sp. caribae... G6 LSU D8-D10 G3 LSU D8-D10 G2 LSU D8-D G4 LSU D8-D10 25 G5 LSU D8-D10 EU Gambierdiscus australes c... EU Gambierdiscus australes c... EU Gambierdiscus australes c EU Gambierdiscus sp. carolin... EU Gambierdiscus sp. carolin... EU Gambierdiscus sp. carolin... 82EU Gambierdiscus polynesiens... EU Gambierdiscus polynesiens EU Gambierdiscus polynesiens EU Gambierdiscus polynesiens... EU Gambierdiscus polynesiens EU Gambierdiscus polynesiens... EU Gambierdiscus sp. ruetzle... EU Gambierdiscus sp. ruetzle... EU Gambierdiscus sp. ruetzle... EU Gambierdiscus sp. ruetzle... EU Gambierdiscus sp. ruetzle EU Gambierdiscus yasumotoi c... EU Gambierdiscus yasumotoi c... EU Gambierdiscus yasumotoi c... EU Gambierdiscus yasumotoi c EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo... EU Gambierdiscus toxicus clo... G. carpenteri G. caribaeus Gambiediscus.sp. Ago Bay (5 strains) G. australes G. carolinianus G. polynesiensis G. ruetzleri G. yasumotoi G. pacificus G. toxicus G. belizeanus
11 Aim of this study To evaluate future risk of CFP in temperate regions, it is necessary to understand the population dynamics of Gambierdiscus sp. This study aimed to reveal the seasonal change in abundance of Gambierdisucs sp. in Ago Bay. But no ecological studies on Gambierdiscus have yet been done in temperate regions.. In addition, the effect of temperature on the growth of Gambiediscus sp. isolated from the bay was also investigated in this study. Temperate regions
12 Study site Ago Bay Google map Mie Kii Peninsula Google map
13 Seaweed bed
14 Depth: m Sampling period November 2008 March 2011 ( once a month or more in warmer seasons) Various macroalgal species were collected by snorkeling and sealed within plastic bags while underwater.
15 Examples of macloalgae collected Chlorophyceae Phaeophyceae Phodophyceae Codium fragile Padinaarborescens Amphiroazonata Sargassumthunbergii Gelidiumelegans
16 Procedure to quantify Gambierdiscus sp. 200 m Shake vigorously 250 times to detach the cells from the algal surface 20 m
17 Procedure to quantify Gambierdiscus sp. 200 m Shake vigorously 250 times to detach the cells from the algal surface 20 m Preserved in 15 ml of 1 % formalin filtered seawater
18 Under the light microscope
19 Under the epifluorescent microscope The cells were stained with calcofluor.
20 Procedure to quantify Gambierdiscus sp. 200 m Shake vigorously 250 times to detach the cells from the algal surface 20 m 15 ml of 1 % formalinfiltered seawater Cell counting Calcofluor staining The abundance was expressed in cells g 1 alga wet weight
21 Growth experiments Effects of temperature on the growth Experimental conditions Temperature 10, 15, 20, 25, 30, 32, 35 Medium f/2 Light intensity 150 mol m 2 s 1 Photo cycle 12h 12h Growth rate divisions day 1 was calculated.
22 Effect of temperature on the growth 32 No growth but No growth & dead
23 Cell densities of Gambierdiscus sp. attached to each different macroalga Nov., 2008 Dec. Jan., 2009 Feb. Mar. Apr. May Jun. Cell density (cells g 1 alga) Jul. Aug. Sep. Sep. Oct. Oct. Nov. Nov. Dec. Jan., 2010 Feb. Mar. Apr. May Jun. Jul. Aug. Sep. Oct. Jan., 2011 Dec. Feb. Mar.
24 Cell densities of Gambierdiscus sp. attached to each different macroalga Nov., 2008 Dec. Jan., 2009 Feb. Mar. Apr. May Jun. Cell density (cells g 1 alga) Jul. Aug. Sep. Sep. Oct. Oct. Nov. Nov. Dec. Jan., 2010 Feb. Mar. Apr. May Jun. Jul. Aug. Sep. Each color corresponds to each different algal species Dec. Jan., 2011 Feb. Mar. Oct.
25 Cell densities of Gambierdiscus sp. attached to each different macroalga Nov., 2008 Dec. Jan., 2009 Feb. Mar. Apr. May Jun. Cell density (cells g 1 alga) Jul. Aug. Sep. Sep. Oct. Oct. Nov. Nov. No preference of Gambierdiscus sp. to attach to macroalga Dec. Jan., 2010 Feb. Mar. Apr. May Jun. Jul. Aug. Sep. Oct. Jan., 2011 Dec. Feb. Mar.
26 Artificial seaweed experiment Cloth (Cotton: 65% Chemical fiber: 35%)
27 Cell densities of Gambierdiscus sp. attached to each different macroalga Nov., 2008 Dec. Jan., 2009 Feb. Mar. Apr. May Jun. Cell density (cells g 1 alga) Jul. Aug. Sep. Sep. Oct. Oct. Nov. Nov. Dec. Jan., 2010 Feb. Mar. Apr. May Jun. Jul. Aug. Sep. Oct. Jan., 2011 Dec. Feb. Mar.
28 Seasonal change in mean abundance ofgambierdiscus sp. attached to the macroalgae ND ND ND ND N D 2008 J F M A M J J A S O N D J F M A M J J A S O N D J F M 2011 ND: Not detected
29 Seasonal change in mean abundance ofgambierdiscus sp. attached to the macroalgae In summer and autumn ND ND ND ND N D 2008 J F M A M J J A S O N D J F M A M J J A S O N D J F M 2011 ND: Not detected
30 Seasonal change in mean abundance ofgambierdiscus sp. attached to the macroalgae In winter ND ND ND ND N D 2008 J F M A M J J A S O N D J F M A M J J A S O N D J F M 2011 ND: Not detected
31 Effect of temperature on the growth 32 No growth but No growth & dead
32 Effect of temperature on the growth Some cells could survive at least for 3 months at 10 C 32 No growth but No growth & dead
33 Seasonal change in mean abundance ofgambierdiscus sp. attached to the macroalgae Overwintering cells! ND ND ND ND N D 2008 J F M A M J J A S O N D J F M A M J J A S O N D J F M 2011 ND: Not detected
34 JAPAN Kuroshio Current Pacific Ocean
35 Seasonal change in mean abundance ofgambierdiscus sp. attached to the macroalgae Gambierdiscus sp. can maintain their population annually in Ago Bay ND ND ND ND N D 2008 J F M A M J J A S O N D J F M A M J J A S O N D J F M 2011 ND: Not detected
36 Sea of Japan Pacific Ocean Sites ( ) show where the presence of the genus Gambierdiscus has been reported in temperate regions.
37 No reports of the presence of Gambierdiscus from the coast in Sea of Japan Sea of Japan Pacific Ocean Sites ( ) show where the presence of the genus Gambierdiscus has been reported in temperate regions.
38 Sea of Japan Tsushima Warm Current JAPAN Kuroshio Current Pacific Ocean
39 Sea of Japan Tsushima Warm Current JAPAN Kuroshio Current Pacific Ocean
40 Hatayama et al. 2011: Nippon Suisan Gakkaishi Sea of Japan Tsushima Warm Current JAPAN Kuroshio Current Gambierdiscus sp. was found in Wakasa Bay. Pacific Ocean
41 Gambierdiscus sp. (spp.) may build autochthonous populations widely along the Japanese coast in temperate regions Sites ( ) show where the presence of the genus Gambierdiscus has been reported in temperate regions.
42
43 First report of occurrence from Shimoda (Hara & Horiguchi 1982)
44 Increase of water temperature would induce Gambierdiscus sp. (spp.) to build larger populations in temperate regions and also spread their distribution toward north
45 Maximum cell density of Gambiediscus sp. in Ago Bay Sargassum thunbergii (Phaeophyceae) Octber 1 st, cells g 1 alga
46 Gambierdiscus spp. abundance in other regions
47 Gambierdiscus spp. abundance in other regions
48 NOW Further studies on ecology and physiology, including toxicology, of Gambierdiscus sp. are certainly needed to evaluate future risk of CFP, even in temperate waters of Japan.
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