Yutaka Okumura (Tohoku National Fisheries Research Institute), Hiroto Ota (Miyagi Prefecture Fisheries Technology Institute, Japan),

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Post-tsunami oyster feeding environment in Miyagi prefecture Yutaka Okumura (Tohoku National Fisheries Research Institute), Hiroto Ota (Miyagi Prefecture Fisheries Technology Institute, apan),

Purpose Temporal trend of oyster production in Miyagi Prefecture 70000 weight with shell (t) 52500 35000 17500 Tsunami ul.-dec. a.-un. a.-dec. Post-tsunami oyster harvest levels (about 20,000 tons/year with shell) are higher than just after-tsunami level (5,000 tons/year). 0 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 year Although the levels are lower than pre-tsunami levels (40,000 60,000 t/year), oyster stocks within the Miyagi Prefecture have gradually recovered. To aid recovery of oyster production, it was necessary to investigate the feeding environment and estimate a suitable stock density for the region.

Tsunami in closed bay, Matsushima Bay (Backwash of second wave, about 1min) Matsushima Bay This video was taken from the laboratory. 8 6 4 2 0 Ishinomaki Higashishiogama Tsunami height(m) Shiogama Matsushima Rifu in Matsushima From Google Map Shichigahama Sendai.M.A (2011) Although the height of the tsunami was lower inside the closed bay than outside of it, numerous recreational boats and bamboo pieces used in Porphyra culture were swept out. The tsunami s influence on the aquaculture industry within the bay quickly became a concern.

Research papers on the influence of tsunami to Area Methods Target After tsunami Data before tsunami References Kesennuma Bay Ofunato Bay phytoplankton, which is main diet of oyster Microscopic observation dinoflagellates increase cysts No Pigment profile biomass no change Yes Microscopic observation Nishitani et Okumura et (2013) (2015) dinoflagellates increase cysts Yes Kaga et (2012) increase Okumura et Pigment profile biomass Yes (2015) biomass Okumura et Dna sequence all species Diversity change Yes (2017) Microscopic low Tachibana et Otsuchi Bay dinoflagellates No (2017) observation concentration the same area. Microscopic Kamiyama et Sendai Bay dinoflagellates increase cysts Yes (2014) observation? Microscopic Watanabe et Many researcher diatom no change Yes (2017) observation have investigated specific phytoplankton and biomass. While, Microscopic Taniuchi et the research of phytoplankton phenology assemblages no change is limited. We Nopresumed the reason (2017) observation The present or absent of the tsunami effect on phytoplankton differ, in spite of why present or absent of the tsunami effect on phytoplankton differs in the same area is the difference of target phytoplankton species. In this study, we compared phytoplankton biomass and phytoplankton diversity before and after the tsunami in Sendai Bay.

Sampling sites Miyagi Prefecture epicenter Nagatsuraura Matsushima Oginohama Sendai Bay We investigate the dietary environment of oysters in Oginohama Bay (HPLC), and the phytoplankton diversity in Sendai Bay (microscopic observation).

Temporal trend of pigments Maximum of pigment contents (before 2011 2012 2013 2014) In Oginohama, spring bloom was observed. Chl a and many pigment concentrations were high after the tsunami. The contour figure of Fuco was similar to Chla. Diatom was predominant taxa.

Phytoplankton diversity was different before and after the tsunami. After the tsunami, phytoplankton diversity varied with the periods. Phytoplankton diversity in Sendai Bay Similarity of phytoplankton data by microscopic observation using nmds The comparison of before (1985-1990) Only after the tsunami and after the tsunami ul2011 May2011 May2011 No2011 ul2011 May2011 before after Data from 1985 to 1990 were published by Sendai city. Ap-May2012 a2012 in Sendai and Oshika a2012 in Mastushima

Calculation method of the suitable culture density of oyster We defined the suitable oyster culture quantity as phytoplankton quantity in the aquaculture areas minus the total filtration, and that value is more than zero. Phytoplankton biomass X Phytoplankton growth rate - Equations of filtration rates by oyster and the periphyton species calculations (F) D.W./ W.W. conditions references Crassostrea gigas 0.26e (0.1584xT) x WW T<11.8 Okumura personal communication Mytilus galloprovincialis (0.70 x T - 6.6) x WW 11.8 < T<18.5 Akashige et (2005) 4.9 x WW 18.5 < T Akashige et (2005) 6.773 DW 0.678 Riisgard et (2014) 5(%) Miura et (2013) Ascidiella aspersa 54.4 DW 1.05 /1000 60 5(%) conversion from ml/min Randlov and Riisgard (1979) Ciona intestinalis 46.4 DW 0.84 /1000 60 5(%) conversion from ml/min Styela clava Herdman exp(0.119t-4.54) DW 0.6745 8.5(%) T<29 Kim Y.S. and Moon T.S. (1998) Halocynthia roretzi Halocynthia aurantium weight (kg) (0.1956+0.0182T) DW 0.7978exp( 100 75 50 25 0-0.0273T) 5<T<25 eong W.G. and Cho S.M. (2013) 12.2(%) Yamamoto and Handa (2001) We substituted the calculation of H. roretzi. F: filteration (L/ind./h), DW: dry weight (g), T: temperature ( ), WW: wet weight Biomass May Bivalves Oyster Total filtration was calculated by multiplying total biomass by filtration rate. X August November Ascidiacea time X February Phytoplankton growth in the culture area was calculated by multiplying Chl a concentration by average growth rate. Total quantity

Estimation of suitable culture density phytoplankton biomass (kg/day/oginohama) 500 350 200 50-100 High culture density post-tsunami Low culture density post-tsunami High culture density pre-tsunami Low culture density pre-tsunami F F M M A A A M M A A S S S O O N N D D F F M Overcrowded farming (<0) Date By current estimations, we believe that a hanging-culture quantity at present condition would not constitute overcrowding. While, we believe that a hanging-culture quantity of high culture density pre-tsunami condition would overcrowding.

Various statistics on oyster culture in Miyagi prefecture The number of companies producing oysters in 2008 and 2013 was 809 and 364, respectively. With the post-tsunami retirement of many employees, and reduction in number of companies, the total quantity of oysters in the aquaculture grounds has decreased. The aquaculture grounds are currently not overcrowded.

Phytoplankton in several samples using Next Generation Sequencing (NGS) To investigate the food of the oysters DNA extraction and purification from seawater, stomach and gut contents of Oyster and Blue mussel PCR of PsbA gene, encoding the D1 protein of photosystem II Mixing PCR products and NGS analys Creation of PsbA gene database from NCBI site and analysis by QIIME bio-pipeline

Heatmap of phytoplankton detected from seawater and shellfishes Number of detection is following order, red > yellow > green > blue Blue mussel (n=76) Oyster (n=55) Seawater(n=49) Stomach gut fecal Stomach gut fecal Diatom (226 species) Chaetoceros socialis Cyclotella meneghiniana Skeletonema_costatum Minidiscus trioculatus Thalassiosira pseudonana inoflagellate Dinophysis spp. Heterocapsa triquetra haeocystis Green algae Ostreococcus sp Bathycoccus prasinos Micromonas pusilla About 450 species were detected. Diatom was predominant taxa. Predominant diatoms were detected from both seawater and shellfishes. Dinoflagellate; Heterocapsa triquetra was mainly detected from shellfishes. Pico-prasinophytes were mainly detected from seawater.

Summary In Oginohama, Chl a and many pigment concentrations were high after the tsunami. In Sendai Bay, phytoplankton diversity was different before and after the tsunami. By current estimations, we believe that a hanging-culture quantity at present condition would not constitute overcrowding. With the post-tsunami retirement of many employees, and reduction in number of companies, the total quantity of oysters in the aquaculture grounds have decreased. We could successfully determine phytoplankton species in seawater and shellfishes using NGS. Predominant diatoms were detected from both seawater and shellfishes. Dinoflagellate; Heterocapsa triquetra was mainly detected from oyster and blue mussel. We thought that the shell of H. triquetra was hard, and could not be digested by shellfishes. Pico-prasinophytes were mainly detected from seawater. We thought that the size of picoprasinophytes were small (3µm), and could not be captured by shellfishes.