Biology & Ecophysiology of ASP toxin producers WESTPAC - - PowerPoint PPT Presentation

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Biology & Ecophysiology

• f ASP toxin

producers

WESTPAC workshop on the development of research strategy for Harmful Algal Blooms

“Toward the future development of HAB science in the Western Pacific – What we know, and what we do not know on HABs”

Institute of Oceanography, Nha Trang, Vietnam 19-22 Dec 2016

Hong Chang LIM

Faculty of Applied Sciences & Computing Tunku Abdul Rahman University College

Sing Tung TENG

Faculty of Resource Science and Technology Universiti Malaysia Sarawak

Suh Nih TAN, Chui Pin LEAW & Po Teen LIM

Bachok Marine Research Station, IOES University of Malaya

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http://harmfulalgae.info

The Pseudo-nitzschia species

• P. abrensis Pérez-Aicua et Orive
• P. americana (Hasle) Fryxell
• P. antarctica Manguin
• P. arctica Percopo & Sarno
• P. arenysensis Quijano-Scheggia, Garcés, Lundholm
• P. australis Frenguelli
• P. batesiana H.C. Lim, Teng, Leaw et P.T. Lim
• P. bipertita Teng, H.C. Lim et Leaw
• P. brasiliana Lundholm, Hasle et Fryxell
• P. caciantha Lundholm, Moestrup et Hasle
• P. calliantha Lundholm, Moestrup et Hasle
• P. circumpora H.C. Lim, Leaw et P.T. Lim
• P. cuspidata (Hasle) Hasle
• P. decipiens Lundholm et Moestrup
• P. delicatissima (Cleve) Heiden
• P. dolorosa Lundholm et Moestrup
• P. fraudulenta (Cleve) Heiden
• P. fryxelliana Lundholm
• P. fukuyoi H.C. Lim, Teng, Leaw et P.T. Lim
• P. galaxiae Lundholm et Moestrup
• P. granii (Hasle) Hasle
• P. hasleana Lundholm
• P. heimii Manguin
• P. inflatula (Hasle) Hasle
• P. kodamae Teng, H.C. Lim, Leaw et P.T. Lim
• P. limii Teng, H.C. Lim et Leaw
• P. linea Lundholm, Hasle et Fryxell
• P. lineola (Cleve) Hasle
• P. lundholmiae H.C. Lim, Teng, Leaw et P.T. Lim
• P. mannii Amato et Montresor
• P. micropora Prissholm et Moestrup
• P. multiseries (Hasle) Hasle
• P. multistriata (Takano) Takano
• P. obtusa (Hasle) Hasle et Lundholm
• P. plurisecta Orive et Pérez-Aicua
• P. prolongatoides (Hasle) Hasle
• P. pseudodelicatissima (Hasle) Hasle emend.

Lundholm, Hasle et Moestrup

• P. pungens (Cleve) Hasle
• P. pungiformis (Hasle) Hasle
• P. roundii Hernandez-Becerril et Diaz-Almeyda
• P. sabit Teng, H.C. Lim, P.T. Lim et Leaw
• P. seriata (Cleve) Peragallo
• P. sinica Qi et Wang
• P. subcurvata (Hasle) Fryxell
• P. subfraudulenta (Hasle) Hasle
• P. subpacifica (Hasle) Hasle
• P. turgidula (Hustedt) Hasle
• P. turgiduloides (Hasle) Hasle

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http://harmfulalgae.info

New species in the region

• P. sinica Qi, Ju & Lei 1994
• P. multistriata (Takano) Takano 1995
• P. circumpora Lim, Leaw & Lim 2012
• P. batesiana Lim, Teng, Leaw & Lim 2013
• P. fukuyoi Lim, Teng, Leaw & Lim 2013
• P. lundholmiae Lim, Teng, Leaw & Lim 2013
• P. kodamae Teng, Lim, Leaw & Lim 2014
• P. sabit Teng, Lim, Leaw & Lim 2015
• P. bipertita Teng, Lim, Leaw & Lim 2016
• P. limii Teng, Lim & Leaw 2016

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http://harmfulalgae.info

Country Species China Indonesia Japan Malaysia Philippines Singapore Thailand Vietnam

• P. abrensis
• + M
• P. americana

+ C, E

• + G, J

+ N

• + G, O
• P. batesiana
• + M, N, P
• P. bipertita
• + M
• P. brasiliana

+ C, D, E + G + J + N, O, Q + T + V + G + G, O

• P. caciantha

+ B, E

• + J

+ N, R + T

• P. calliantha

+ B, E

• + J

+ N, O

• + W

+ O, Y

• P. circumpora
• + O, Q
• + Y
• P. cuspidata

+ B, D, E

• + F

+ Q

• + V
• + O, Y
• P. decipiens
• + N
• P. delicatissima

+ D, F

• + J

+ O

• + W

+ O

• P. dolorosa
• + N, Q
• P. fraudulenta
• + J
• + O
• P. fukuyoi
• + M, P, Q
• + X
• P. galaxiae
• + J
• P. cf. granii
• + O
• P. hasleana
• + N
• + Y
• P. heimii
• + W
• P. inflatula
• + N
• + W

+ O

• P. kodamae
• + M, N, R
• P. limii
• + M
• P. linea

+ C

• + N
• P. cf. lineola

+ E

• + N
• P. lundholmiae
• + M, P
• P. mannii

+ B, E

• + N
• P. micropora
• + N, O

+ T + V + W + O

• P. multiseries

+ E

• + J, K
• P. multistriata

+ D, E

• + J, L

+ M, O

• + V
• + O
• P. pseudodelicatissima

+ A, B, E

• + J
• + T
• + Y
• P. pungens

+ A, D, E + H, I + J + M, Q + T, U + V

• + O, Y
• P. sabit
• + S
• P. cf. sinica
• + W

+ O, Y

• P. sinica

+ B, E

• + N
• P. subpacifica

+ E

• + W
• P. subfraudulenta

+ E

• + M
• + O
• P. turgidula
• + N
• 16

2 12 28 5 5 8 17

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http://harmfulalgae.info

DA producing Pseudo-nitzschia

• P. abrensis

0.0011 pg/cell

• P. australis

0.026-37 pg/cell

• P. batesiana

0.0002 pg/cell

• P. brasiliana

0.0095 pg/cell

• P. cf. caciantha

111-244 ng/mL

• P. calliantha

0.0057-0.22 pg/cell

• P. cuspidata

0.019-0.031 pg/cell

• P. delicatissima

0.0002-0.12 pg/cell

• P. fraudulenta

0.0008-0.03 pg/cell

• P. fukuyoi

3.85-4.54 pg/cell

• P. galaxiae

0.00036 pg/cell

• P. granii

0.000004 pg/cell

• P. kodamae

1.2-42.5 pg/cell

• P. lundholmiae

0.0011 pg/cell

• P. multiseries

0.006-67 pg/cell

• P. multistriata

0.04-0.697 pg/cell

• P. plurisecta

0.00024-0.32 pg/cell

• P. pseudodelicatissima

0.0078 pg/cell

• P. pungens

0.0018-0.47 pg/cell

• P. seriata

0.8-33.6 pg/cell

• P. subfraudulenta

0.0011-0.0023 pg/cell

• P. subpacifica

0.06-1.1 ng/mL

• P. turgidula

0.0000052-0.09 pg/cell

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http://harmfulalgae.info

Toxicity under different conditions

Teng et al. 2014; Updated

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http://harmfulalgae.info

Confirmed toxic species in WESTPAC region

• P. multiseries (1999)

Ofunato Bay, Japan

• P. cf. caciantha (2014),
• P. fukuyoi (2015)

Khan Hoa Bay, Nha Trang, Vietnam

• P. abrensis (2016)
• P. batesiana (2016)
• P. fukuyoi (2016)
• P. kodamae (2016)
• P. subfraudulenta (2016)

Miri, Sarawak, Malaysia

• P. kodamae (2014)

Port Dickson, Malaysia

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http://harmfulalgae.info

Occurences

1. Santubong River and Samariang, Sarawak, Malaysia ‐ peaked during April‐May (2007‐2010) (Lim et al. 2012 MRI) ‐ increase

• f

cell density corresponding to low precipitation, high salinity and pH (no clear trend with changes of macronutrient in the water) (Lim et al. 2012) 2. Philippines and Thailand ‐ higher cell density observed during dry season from Dec‐May (2005‐2006, Yap‐Dejeto et al. 2008; Udomratana et al. 2008) 3. Cisadane River Estuary, Indonesia ‐ Higher cell density during dry season May‐June (2011) ‐ corresponding to changes in salinity and N:P concentrations (Rachman & Thoha 2014 MRI)

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http://harmfulalgae.info

Occurences

• 4. Nha Phu, Binh Cang and Nha Trang waters, Khanh Hoa

Province, Vietnam ‐ density peak during July‐Aug, Nov (2009‐2010) (Hai et al. 2013)

• 5. a. Bohai Bay, China (Yellow Sea, North)

‐ dominant during Summer (P. pungens) (Chen et al. 2016)

• b. Daya Bay, China (South China Sea)

‐ peaks during Spring and Autumn, July 1997‐June 1998 (Chen et al. 2002) ‐ high density during Spring (P. pseudodelicatissima and P. pungens), less grazing pressure from zooplankton (Cheng et

• al. 2005)
• c. Zhelin Bay, China (South China Sea)

‐ May 2000‐Dec 2004, from 214 samples, 20.5% with P. pungens densities > 104 cell/L and 3.8% with > 105 cell/L ‐ water temperature (23.8oC or higher) showed highly significant linear relationship with the algal bloom (Huang et

• al. 2008)

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http://harmfulalgae.info

Cell abundances

29 35 4.5 127 38 293 510 100 200 300 400 500 600

Sarawak, Malaysia San Pedro Bay, Philippines Cisadane River Estuary, Indonesia Cua Be Estuary, Vietnam China [Bohai, Daya Bay, Zhelin Bay]

Cell density 103 Santa Cruz, North American west coast ‐ ~5 x 105 cells/L of P. australis (May 2015, Spring) with 19,978 ng/mL of pDA recorded in California (McCabe et al. 2016)

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http://harmfulalgae.info

Identification – morphological characterization

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http://harmfulalgae.info

TEM

• P. limii

Teng et al. 2016 [J. Phycol.]

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http://harmfulalgae.info

Morpho-characters

1. Transapical axis (cell width) [ > 3 micron or < 3 micron] 2. Center interspace/central nodule (absent/present) 3. Shape of valve (symmetrical? Asymmetrical? Linear? Lanceolate?) 4. Cell at girdle view (sigmoid? rhomboid?) 5. Cell overlap (1/4, 1/8, 1/10?) 6. Shape of apices 7. Density of fibulae in 10µm 8. Density of striae in 10µm 9. Ratio of fibulae vs. striae 10. Height of proximal and distal mantles 11. Number of rows of poroids (1 or 2 or 3 or 4?) 12. Number of poroids in 1µm 13. Structure of poroids (how many dividing sectors?) 14. Shape of poroid 15. Density of band striae/poroids in valvocopula 16. Structure of band striae 17. Chain forming

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http://harmfulalgae.info

3I Interactive Keys and Taxonomic Databases

Teng et al. 2014 (HANS) http://dmitriev.speciesfile.org/key.asp?key=Bacillariales&lng=En&i=1&keyN=2

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http://harmfulalgae.info

Identification – molecular characterization

Lim et al. In Prep

cox1 LSU

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http://harmfulalgae.info

18S secondary structure

Lim et al. 2016 [Phycologia]

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http://harmfulalgae.info 18s/SSU

Identification – molecular characterization

Lim et al. 2016 [Phycologia]

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http://harmfulalgae.info

ITS2 secondary structure

Lim et al. 2013 [J. Phycol.]

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http://harmfulalgae.info

Compensatory base change (CBC)

Teng et al. 2016 [J. Phycol.]

CBC species concept – if there is a CBC, there is a probability of ~ 0.93 that there are two different species (Wolf et al. 2013 PLOS One)

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http://harmfulalgae.info ITS2

Lim et al. In Prep

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http://harmfulalgae.info

• P. cuspidata vs. P. pseudodelicatissima

Lundholm et al. 2003; Lelong et al. 2012; Trainer et al. 2009, 2012

• P. pseudodelicatissima
• P. cuspidata

Linear valve Lanceolate valve

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http://harmfulalgae.info

• P. cuspidata? P. pseudodelicatissima?
• P. cuspidata

Type Locality: Tenerife, Gran Canary Island (most closely represented by strain “Tenerife8”) (Lundholm et al. 2012)

• P. pseudodelicatissima

Type locality: subarctic Denmark Strait, no available strain (Hasle, 1965) Lim et al. In Prep

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http://harmfulalgae.info

• P. cuspidata? P. pseudodelicatissima?

Two possible scenarios remain: 1)

• P. pseudodelicatissima will be

a synonym of P. cuspidata if a representative strains from the holotype area (i.e., Denmark Strait) falls in the same clade as P. cuspidata; 2)

• P. pseudodelicatissima is the

name of another clade if a representative strain from the holotype area appears in another clade.

Lim et al. In Prep

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http://harmfulalgae.info

Other ASP toxin producers

Nitzschia navis‐varingica Lundholm and Moestrup 2000 Nitzschia bizertensis Bouchouicha Smida, Lundhom, Sakka and Hadj Mabrouk 2014 Toxicity:

• N. navis‐varingica 0.37‐11.06 pg/cells (Suriyanti & Usup, 2015;

Tan et al. 2016)

• N. bizertensis

0.0002‐0.036 pg/cells (Smida et al. 2014) Halamphora coffeiformis (Agardh) Levkov 2009 [formerly known as Amphora coffeaeformis] – Only one Canadian strain was found to produce DA. However, identity of the toxigenic species remained uncertain (Sala et al. 2011)

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http://harmfulalgae.info

Nitzschia navis-varingica

Tan et al. 2016, Harmful Algae

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http://harmfulalgae.info

Molecular characterization of Nitzschia navis-varingica

Tan et al. 2016, Harmful Algae

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http://harmfulalgae.info

Nitzschia navis-varingica

Tan et al. 2016, Harmful Algae

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http://harmfulalgae.info

Recommendation/ Future work

a. Understanding on Pseudo‐nitzschia blooms, its versatility relative to other phytoplankton; b. Blooms initiation/ retention of different (planktonic and epiphytic) ASP producers? c. Similar/higher blooms density of Pseudo‐nitzschia than those in US, but no DA accumulation in WESTPAC region? d. Complexity of the taxonomy: taxonomical status of P. cuspidata and P. pseudodelicatissima; e. Taxonomical revision of the genus Nitzschia; f. Clarification of DA production by Halamphora coffeiformis and its’ taxonomical identity; g. Molecular tools for detection, e.g. qPCR; h. DNA barcoding for Pseudo‐nitzschia

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Thank you

http://harmfulalgae.info