Harmful Algae 14 (2012) 71–86
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Harmful Algae
journal homepage: www.elsevier.com/locate/hal
Harmful dinoflagellate blooms caused by Cochlodinium sp.: Global expansion and
ecological strategies facilitating bloom formation
Raphael M. Kudela a,*, Christopher J. Gobler b
a
b
Ocean Sciences Department, University of California Santa Cruz, 1156 High Street, Santa Cruz, CA 95064, USA
School of Marine and Atmospheric Sciences, Stony Brook University, 239 Montauk Highway, Southampton, NY 11946, USA
A R T I C L E I N F O
A B S T R A C T
Article history:
Available online 25 October 2011
The past two decades have witnessed an expansion in the reported occurrences of harmful algal blooms
(HABs) caused by the dinoflagellate Cochlodinium. Prior to 1990, blooms had been primarily reported in
Southeast Asia, with South Korea alone reporting more than $100M USD in annual fisheries losses during
the 1990s. Since then, time blooms have expanded across Asia, Europe, and North America, with
recognition of multiple species and ribotypes that exhibit similar ecophysiological and harmful
characteristics. Here, we summarize the current state of knowledge regarding taxonomy, phylogeny,
detection, distribution, ecophysiology, life history, food web interactions, and mitigation of blooms
formed by Cochlodinium. We review this recent expansion of Cochlodinium blooms and characterize the
ecological strategies utilized by Cochlodinium populations to form HABs. Although Cochlodinium is
comprised of more than 40 species, we focus primarily on the two HAB-forming species, C. polykrikoides
and C. fulvescens, specifically describing their flexible nutrient acquisition strategies, inhibition of grazing
by inducing rapid mortality in a diverse set of predators, and allelopathic inhibition of a broad range of
competing phytoplankton. Finally, we summarize the available information on prevention, control, and
mitigation strategies specific to this genus, and discuss pressing questions regarding this increasingly
important HAB organism.
ß 2011 Elsevier B.V. All rights reserved.
Keywords:
Cochlodinium polykrikoides
Cochlodinium fulvescens
Ecophysiology
Food web interactions
Harmful algal bloom
Mitigation
1. Introduction
Dinoflagellates of the genus Cochlodinium were first identified
in 1895 by Schütt (1895) and have been forming harmful algal
blooms in the coastal waters of Southeast Asia and North America
for many decades. The past two decades have seen Cochlodinium
blooms expanded in their geographic distribution across Asia,
Europe, and North America (Fig. 1), with fisheries losses associated
with blooms in South Korea alone exceeding $100M annually (Kim,
1997). More than 40 species of Cochlodinium have been described,
although the two primary HAB-forming species are C. polykrikoides
and C. fulvescens (Fig. 2). Both of these species are large (40 mm)
athecate dinoflagellates that commonly form chains of 2–16 cells.
Cochlodinium blooms are generally characterized by spatially large
(10s to 100s of kilometers) and dense (>1000 cells ml 1) cell
aggregates that are heterogeneous in their vertical and horizontal
distributions. These blooms are strongly ichthyotoxic and can also
kill many other marine organisms, although the compound(s)
responsible for these impacts have yet to be identified and bloomassociated toxins are not known to affect human health. Partly due
* Corresponding author. Tel.: +1 831 459 3290; fax: +1 831 459 4882.
E-mail address: [email protected] (R.M. Kudela).
1568-9883/$ – see front matter ß 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.hal.2011.10.015
to the recent expansion of Cochlodinium blooms and the general
difficulty in culturing this species, there is far less known about the
autecology and toxicity of Cochlodinium compared to other HAB
species, particularly for the recently described C. fulvescens. With
this review we seek to characterize the current state of knowledge
regarding taxonomy, phylogeny, detection, distribution, ecophysiology, life history, food web interactions, and mitigation of blooms
formed by Cochlodinium as well as point out pressing questions
regarding this increasingly important HAB genus.
2. Taxonomy and phylogeny
As with many of the unarmored dinoflagellates, morphological
features vary as a function of environmental conditions and life
cycle. Typical preservation methods can distort or even destroy
cells (Fig. 4), leading to under-representation and mis-identification of organisms with morphology similar to Cochlodinium such as
Gymnodinium catenatum and Gymnodinium impudicum (e.g. Cho
and Costas, 2004; Curtiss et al., 2008; Howard et al., in press). It is,
perhaps, not surprising that the taxonomy of even the most widely
studied Cochlodinium species is unclear. The genus Cochlodinium
was established more than a century ago with the identification of
C. strangulatum (Schütt) Schütt (1895). The majority of reported
organisms within this genus (50 species, with 40 accepted; Guiry
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R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
Fig. 1. Global distribution of reported Cochlodinium events showing the apparent expansion in blooms before (top panel) and after (bottom) 1990.
and Guiry, 2011) are rare heterotrophic organisms, and not well
studied or described. Within this large group, only four species are
known to produce chloroplasts and form chains: C. polykrikoides
(synonymous with C. catenatum Okamura, 1916 and C. heterolobactum Silva, 1967), C. fulvescens, C. geminatum, and C.
convolutum (Iwataki et al., 2007; Matsuoka et al., 2008). Of those
species, only two, C. polykrikoides and C. fulvescens, are confirmed
ichthyotoxic organisms (Fig. 2).
The holotype for C. polykrikoides was originally described by
Margalef (1961) based on isolates from Puerto Rico, although
there is some dispute about the proper naming of the species since
C. catenatum Okamura, 1916 was described first, with the two
organisms now accepted to be sub-clades of the same species (c.f.
Matsuoka et al., 2008). The genus is almost certainly polyphyletic
(Iwataki et al., 2010) with the sole diagnostic character being that
the cingulum surrounding the cell more than 1.5 times (Kofoid
and Swezy, 1921; Iwataki et al., 2007, 2010). Besides the
characteristic cingulum encircling the cell twice, C. polykrikoides
has a reddish orange eyespot in the anterior dorsal part (epicone)
of the cell, with multiple rod-like chloroplasts. As described, the
holotype is 50 mm in length, and typically forms chains of up to
16 cells in length. C. catenatum Okamura, 1916 was described as a
smaller (21–26 mm in length), chain-forming dinoflagellate with
the cingulum encircling the cell more than 1.5 times, with a
smaller eyespot located in the epicone, and with multiple rod-like
chloroplasts. C. catenatum Kofoid and Swezy, 1921, is superficially
similar to C. catenatum sensu Okamura but was described as
heterotrophic with a more central nucleus. A third organism, C.
convolutum Kofoid and Swezy, 1921 is larger (60–70 mm in
length), lacks an eyespot, has reticulate chloroplasts together with
many scattered small grains at the periphery of the cell, and rarely
forms chains. C. convolutum has been reported to form red tides
(discolorations of the surface water) in both Japan and California,
but has not been implicated in fish or shellfish mortality (c.f.
Matsuoka et al., 2008). The most recently described species in this
genus, C. fulvescens Iwataki, Kawami and Matsuoka, 2007, is 40–
50 mm in length, forms chains of 2–4 cells (up to 8 cells have been
observed; Kudela, pers. obs.), has a cingulum that wraps 2
around the cell (similar to C. polykrikoides), has a reddish eyespot
in the dorsal epicone, and exhibits granular chloroplasts (Table 1).
This organism has been difficult to culture and there is relatively
little known about its ecophysiology or the mode of action for
harmful effects, but blooms in British Columbia, Canada and
central California have been linked to mortality of aquacultured
salmon and abalone (Whyte et al., 2001; Kudela, unpublished
data).
Based on the recent morphological characterization of C.
polykrikoides and C. fulvescens (Iwataki et al., 2007, 2008, 2010;
Matsuoka et al., 2008), there appear to be enough significant
differences with morphologically similar Gymnodinium species to
maintain the Cochlodinium genus despite the polyphyletic nature
of the group. Shao et al. (2004) demonstrated that for a single strain
of C. polykrikoides there was good evidence for separation from
Gymnodinium, Amphidinium, Gyrodinium, Karlodinium, and Karenia,
using internal transcribed spacer (ITS) rDNA sequences. C.
polykrikoides was placed within the gymnodinoid, peridinioid,
prorocentroid complex (GPP; Saunders et al., 1997), and formed a
clade with sequences from Pfisteria piscicida and Akashiwo
sanguinea rather than the morphologically similar Gyrodinium
and Gymnodinium genera. More recently, Ki and Han (2008) fully
sequenced the large subunit (LSU) rDNA from four Korean strains
of C. polykrikoides, and determined that the core DNA sequences are
highly conserved and therefore amenable to use for long-term
evolutionary phylogeny of dinoflagellates, while the hypervariable
D1–D3 region is best suited for species or strain-level differences.
Iwataki et al. (2007, 2008) also used LSU rDNA sequences to
demonstrate that C. fulvescens is closely grouped with, but is a
sister clade of C. polykrikoides, supporting the separation as a new
species based on cell morphology. Those authors, however,
identified three distinct sub-clades, or ribotypes, in C. polykrikoides; the East Asian ribotype (also referred to as Japanese–
Korean), the American/Malaysian ribotype, and the Philippines
ribotype. More recently, another morphotype called ‘‘Kasasa’’ has
been proposed on the basis of morphology, size, and formation of
2-cell chains (Matsuoka et al., 2010), but has so far not been
sequenced and is very similar to the Japanese–Korean ribotype as
described. The primary distinguishing characteristics between C.
fulvescens and C. polykrikoides ribotypes are the relative position of
cingulum and sulcus, and the morphology of chloroplasts (Table 1).
Based on additional LSU sequences, it is possible that C. fulvescens
will also be divided into multiple ribotypes (Fig. 3).
To summarize, there is good morphological and phylogenetic
evidence for the establishment of two ichthyotoxic, bloomforming members of the Cochlodinium genus: C. polykrikoides
and C. fulvescens. To date, LSU sequences and morphological
descriptions for most other species of Cochlodinium have been
somewhat limited relative to the global distribution of blooms (see
Section 4). Enhanced sequencing efforts in regions where blooms
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
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Fig. 2. Line drawings of Cochlodinium fulvescens (A) and C. polykrikoides (B).
have newly formed but have been poorly described may resolve
current uncertainties regarding evolutionary relationship, phylogeny, and population origin and transport. Within one of these
species there are at least three (possibly four) sub-clades or
ribotypes of C. polykrikoides (Fig. 3). What is still lacking is a
clear phylogenetic relationship between Cochlodinium and other
dinoflagellate genera, and a clear molecular basis for the inclusion
of 40–50 species within the genus. Recent work (e.g. Ki and Han,
2008) suggests that basing the evolutionary history on the LSU
sequences should be feasible, but would require more full
sequences from other dinoflagellates. Similarly, a stable taxonomy for the genus Cochlodinium will likely require a careful
Table 1
Morphological characteristics of the most commonly reported photosynthetic species of Cochlodinium.
Size
Cingulum
Chloroplasts
Pigmented body
Chain formation
C. polykrikoides
C. fulvescens
C. catenatum
C. convolutum
50 mm
Encircling >1.5
Rod-like
Present, dorsal epicone
>4 cells
40–50 mm
Encircling 2
Granular
Present, dorsal epicone
>4 cells
21–26 mm
Encircling >1.5
Rod-like
Present, dorsal epicone
>4 cells
60–70 mm
Encircling 1.5
Small, scattered
Absent
Rarely formed
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Fig. 3. Phylogenetic relationships based on LSU rDNA were inferred using the Neighbor-Joining method with bootstrap support values shown (as a percentage of 500
replicates). The tree is to scale with branch lengths proportional to genetic distance (Maximum Composite Likelihood method) between aligned sequences. Positions
containing alignment gaps and missing data were excluded in pairwise sequence comparisons.
Fig. 4. Light micrographs of Cochlodinium. (A) C. fulvescens obtained from a red tide event in coastal waters of Avila Beach, California (USA). (B) C. polykrikoides isolated from
estuarine waters of the US east coast; (C) C. fulvescens from Avila Beach demonstrating the effect of formalin preservation on cell morphology. Scale bars in (A) and (C) are
50 mm.
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
reexamination of the type organism C. strangulatum originally
described by Schütt (1895) and molecular characterization and
more detailed morphological observations for other members of
the genus.
3. Detection
The detection of Cochlodinium can be divided into three general
categories: microscopy, molecular techniques, and remote sensing
(in the broadest sense of the term). The large size and
distinguishing features of many species of Cochlodinium cells
generally make them a group of HABs that is easy to identify and
quantify microscopically, particularly when populations have
achieved bloom densities. Microscopic quantification of Cochlodinium populations during the earliest stage of blooms or in
sediments, however, is highly challenging, particularly in the case
of post-preservation identification, necessitating alternative
approaches such as molecular techniques for cell detection. The
characteristic, large, dense (>1000 cells ml 1) surface aggregates
formed by Cochlodinium blooms make them highly amenable to
both optical and acoustic remote detection methods.
3.1. Molecular tools
Given the difficulty of identifying many HAB organisms to
species level and the problematic preservation of cells using typical
methods for naked dinoflagellates, many research groups have
turned to the molecular toolbox for identification and quantification (c.f. reviews by Humbert et al., 2010; Kudela et al., 2010). A
common molecular approach for cell identification is fluorescent in
situ hybridization (FISH), also known as Whole Cell Probing (WCP).
This method employs synthesized ‘‘probes’’ with a covalently
linked fluorescent dye, or reporter marker, usually targeting highly
abundant ribosomal RNAs (rRNAs). Early identification of Cochlodinium was hampered by the paucity of gene sequences available
for the development of species- or ribotype-specific probes. To
overcome this, FITC-labelled lectins were utilized (Costas and
Rodas, 1994). Lectins bind non-covalently to simple sugars and
polysaccharides; while binding is not specific to any one species or
group, the wide availability of multiple lectins allows for a matrixbased approach capable of distinguishing morphologically similar
species, such as C. polykrikoides, G. impudicum, and G. catenatum,
without the necessity of sequence data (Cho et al., 1998; Cho, 2002;
Cho and Costas, 2004).
The increasing availability of sequence information within the
last decade has resulted in widespread adoption of more specific
antibodies or rRNA/rDNA markers (Cho and Costas, 2004; Mikulski
et al., 2008). Closely related to whole-cell probing are cell
homogenate methods, which add a chaotropic compound to the
sample, thereby releasing the rRNA (or other targets) from the cell.
These variants include sandwich hybridization assay (SHA) and
polymerase chain reaction (PCR) methods (reviewed by Humbert
et al., 2010; Kudela et al., 2010). Mikulski et al. (2008) reported the
development of both WCP and SHA using the D1–D3 region of the
LSU from 8 strains of C. polykrikoides and 1 strain of C. fulvescens
(thought at the time to be C. polykrikoides), representing 6 strains
from the Japanese–Korean ribotype and 2 strains from the
American/Malaysian ribotype. Specificity and detection limit were
reported to be very good, with a SHA limit of detection equal to
115 cells ml 1 lysate, and no reported cross-reactivity for a range
of other dinoflagellates (Mikulski et al., 2008).
Potential concerns for these methods include cross-reactivity
with non-target organisms, non-specific labelling of organic
material, and lack of reactivity due to genetic divergence within
the probe/primer region. When Cochlodinium recently emerged in
central California (ca. 2004; Curtiss et al., 2008; Kudela et al., 2008),
75
preliminary microscopic observations identified it as either C.
polykrikoides or C. catenatum Kofoid and Swezy, 1921 (Fig. 4).
Because of the similar morphological characteristics (particularly
after preservation; Matsuoka et al., 2008) and the documentation
of a complex life cycle in C. polykrikoides including armored,
unarmored, and resting cyst stages (Kim et al., 2007), the WCP
probes of Mikulski et al. (2008) were applied to the unknown field
samples. No cell-specific labelling occurred for the California
samples, although there was some cross-reactivity with cooccurring strains of Alexandrium catenella. Using single-cell PCR,
partial LSU rDNA (D1–D6) sequences were obtained (Iwataki et al.,
2008) which identified the California strain as C. fulvescens.
Identification of the new ribotype C. fulvescens also places the
strain from British Columbia (used by Mikulski et al., 2008) into
this same group (Fig. 3); thus the oligonucleotide probes developed
for C. polykrikoides inadvertently included sequence information
from a closely related, but genetically distinct, organism (C.
fulvescens), resulting in what may more accurately be characterized as a potentially multi-species-specific probe.
In addition to WCP and SHA, real-time quantitative PCR (qPCR)
methods for C. fulvescens and C. polykrikoides have been recently
developed (Park et al., 2009; Howard et al., in press). Both groups
targeted the 18S, or small subunit (SSU) rDNA of the respective
organisms. Park et al. (2009) further applied a post-PCR method,
melting curve analysis which determines the temperature at
which the PCR amplicons melt. This gives a quantitative estimate
of the GC content and the absolute order of the bases in the
amplicon sequence, providing more specificity between species or
strains while maintaining the high throughput advantages of qPCR
(Park et al., 2009). Both groups reported excellent specificity and
good sensitivity with dynamic ranges covering 7–10 orders of
magnitude for predicted cell abundances. Both groups also
reported good agreement between qPCR enumeration methods
and traditional microcscopy from field samples. Such approaches
may prove valuable for assessing the earliest stages of bloom
formation (e.g. <10 cells ml 1), a phenomenon that has been
intractable via traditional, microscopic approaches, and thus is
poorly understood.
Park and Park (2010) further extended the use of real-time qPCR
by analyzing sediment samples for the presence of both C.
polykrikoides and G. impudicum in Korean coastal samples. They
were unable to attribute the presence of nucleic acids to any
specific life stage, however, leading to the possibility that they
detected intact cells, resting stages, cysts, or degraded cellular
material. Real-time PCR of environmental samples, particularly
sediment samples, is potentially subject to several sources of error
including inhibition due to (e.g.) humic acids, phenolics, and heavy
metals, cross-reactivity with non-target species, and identification
of dead vegetative cells. Most PCR methods report concentrations
in units of amplicon copy number. To convert this copy number to
cell densities, it is also necessary to know (or estimate) the copy
number per cell of the target sequence. For C. polykrikoides, the
rDNA content per cell is currently unknown, and little is known at
this time regarding cyst formation in this species, ruling out a
direct calibration of the PCR method with known cyst densities.
Despite these issues, Park and Park (2010) demonstrated that realtime qPCR could be used to detect relative abundances (copy
number) and geographic distribution of C. polykrikoides sequences
in coastal sediments. This in turn provides a useful molecular
approach for identifying benthic populations.
To date, rDNA methods such as WCP, SHA, and real-time qPCR
have proven ideal for identification and potential quantification of
species or strains of C. polykrikoides as well as the presence or
absence of vegetative cells and/or resting stages in sediments. The
limited efforts to sequence C. polykrikoides populations have
uncovered relatively little genetic variability (Ki and Han, 2008)
76
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and there is insufficient information within the hypervariable SSU
or LSU regions to assess questions about gene flow or ballast water
transport (Nagai et al., 2009). An increasingly popular molecular
technique that can be used to address these fine-scale questions
involves analysis of microsatellites, 1–4 base pair non-encoding
repeating sequences found in nuclear or organelle DNA. Nagai et al.
(2009) have applied this to C. polykrikoides to assess population
expansion within the Sea of Japan. The authors concluded that
expansion of the population into the Sea of Japan was likely
associated with physical transport within the Tsushima Warm
Current from the Korean coast, that three sub-populations
currently exist in the region, and that there is some evidence for
human-mediated transport between the coastal Japanese cities of
Nagasaki, Hyogo, and Mie. While limited in geographic extent, this
study demonstrates the potential utility of microsatellite-based
approaches for understanding the rapid apparent global expansion
of Cochlodinium within the past few decades.
3.2. Acoustic methods
An innovative method for the real-time, indirect (remote)
detection of C. polykrikoides takes advantage of acoustical
backscatter, similar to the use of acoustics for identifying and
tracking zooplankton (Holliday et al., 1989), but necessitating the
use of much higher frequencies (more power) to compensate for
the small target size. Two groups have developed this method,
using 5 and 10 MHz ultrasound transducers with both lab and field
trials (Bok et al., 2010; E. Kim et al., 2010). Acoustical methods
appear promising for the early detection of dinoflagellate bloom
initiation. E. Kim et al. (2010) reported a rapid increase in signal
when dinoflagellate concentrations exceeded 300 cells ml 1 while
Bok et al. (2010) reported from a combination of laboratory and
field trials the feasibility of detecting between 300 and
1000 cells ml 1, equivalent to ‘‘precaution’’ and ‘‘warning’’ stages
for Korean coastal waters (NFRDI 2004 as reported in Bok et al.,
2010). Primary limitations include the inability to differentiate
dinoflagellate species, potential interference from suspended
particulate matter, and the paucity of laboratory and field data.
3.3. Optical remote sensing
Cochlodinium is a relatively large, motile dinoflagellate that
exhibits diel vertical migration and is generally associated with a
relatively restricted range of temperatures and salinities, making
it amenable to remote sensing approaches. Cochlodinium blooms
are typically concentrated at the sea surface at mid-day
(coincident with satellite overpasses) and are frequently associated with mesoscale physical properties (temperature, salinity
and currents). For the specific case of identifying and tracking
Cochlodinium blooms, much work has been done in Southeast Asia
(particularly Korea) and to a lesser extent in North America. These
efforts can be loosely categorized as either (1) using remote
sensing data to provide environmental context to bloom dynamics, or (2) developing specific remote sensing products to identify
red tides of Cochlodinium. Both approaches have merit as a key
first step towards the use of remote sensing data for a long-term
monitoring program is to develop a heuristic model of when
blooms are likely to occur based on knowledge of the regional
bloom characteristics, oceanography, and environment (Stumpf
and Tomlinson, 2005).
Based on a combination of satellite chlorophyll, sea surface
temperature, current, and wind patterns, extreme blooms of
Cochlodinium often appear to be associated with initiation and
subsequent transport from offshore or upstream of coastal sites
(e.g. Ahn et al., 2005, 2006; Azanza et al., 2008; Kudela et al., 2008;
Shanmugam et al., 2008; Nagai et al., 2009; Onitsuka et al., 2010),
consistent with the concept of a ‘‘pelagic seed bank’’ (c.f. Kudela
et al., 2008). Remote sensing can be particularly useful for
identification of the initiation and transport sites, which are often
associated with distinct temperature and salinity properties (e.g.
B.-K. Lee et al., 2008), and provides an opportunity to identify,
track, and therefore forecast bloom impacts on coastal regions.
Progress has also been made on developing Cochlodiniumspecific satellite products. Palanisamy et al. (2005) and Ahn et al.
(2006) attempted to identify blooms by using a statistical
classification scheme comprising Forward Principal Component
Analysis and Minimum Spectral Distance (FPCA and MSD)
methods. Briefly, the remote sensing data are statistically classified
into various groups using known (training) data, and then
unknown data are mapped to the same classifiers. Similar
approaches have been used successfully for other HAB organisms
(e.g. Babin et al., 2005), and this is an active area of research within
the remote sensing community. The analysis of Palanisamy et al.
(2005) suggested that a statistical approach is feasible for detecting
Cochlodinium in specific regions, but is limited by the spatial
resolution (1 km) of most ocean color data. To address this
problem, Ahn et al. (2006) applied the same methods to Landsat-7
ETM+ data, which has greatly increased spatial resolution but
decreased spectral resolution and very poor temporal resolution.
The authors concluded that it is possible to identify Cochlodinium
blooms specifically, but that complex waters and patchy spatial
distributions are problematic.
More recently, Maldonado (2008) performed a thorough
characterization of the optical properties of C. polykrikoides using
both lab (pure culture) and field samples in southwestern Puerto
Rico. That study concluded that Cochlodinium is optically similar
to many co-occurring dinoflagellates, making it difficult or
impossible to identify C. polykrikoides specifically, consistent
with previous studies of other red tide organisms (Dierssen et al.,
2006). As many others have concluded, remote sensing in
combination with in situ verification of the causative organisms
and modeling shows great promise for detecting and tracking high
biomass algal blooms such as Cochlodinium. Several groups have
pursued this nested approach wherein satellite remote sensing is
used with in situ verification to detect the presence or transport of
high-biomass, dinoflagellate blooms (red tides). Red tides can be
identified by quantifying either the near-infrared (NIR) peak in
upwelling radiance caused by surface aggregation of high biomass
(Gower et al., 2005) or the shift towards the red in peak reflectance
(Dierssen et al., 2006; Ryan et al., 2009). While only one multispectral satellite sensor, MERIS, has been specifically designed to
measure NIR signal for computation of an extreme bloom index,
termed the Maximum Chlorophyll Index (MCI; Gower et al., 2005;
Gower and King, 2007), similar algorithms have been developed
specifically for application to Cochlodinium blooms using in situ
optical data (Sasaki et al., 2008), MODIS (Kim et al., 2009), and
SeaWiFS (Shanmugam et al., 2008). These specialized products
overcome limitations inherent to standard (‘‘global’’) products
which often mask red tides as ‘‘bad’’ data but are limited to high
biomass surface blooms, providing little or no information about
bloom initiation or high cell concentrations below the first optical
depth (approximately the upper third of the euphotic zone). Thus
while remote sensing methods are extremely useful for identifying and tracking blooms, an integrated ocean observing system
combining multiple detection and tracking methods is still
required for both monitoring and research purposes (Jochens
et al., 2010).
4. Distribution and expansion of Cochlodinium blooms
Many investigators have noted the global expansion of HABs
during the past several decades with regard to their impacts,
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
duration, intensity, and/or distribution (Anderson, 1989; Hallegraef, 1993; Smayda, 1997; Anderson et al., 2002; Glibert et al.,
2005). While Cochlodinium has been known to science for more
than a century (Schütt, 1895), there has been a remarkable
expansion in the reported distribution of the cells and blooms of
this genus and C. polykrikoides as a species during the past two
decades across Asia, North America, Australia, and even Europe
(Fig. 1). Prior to 1990, C. polykrikoides blooms had been reported as
isolated occurrences in Southeast Asia (Japan, Korea; Yamatogi and
Maruta, 2002; Yuki and Yoshimatsu, 1989) and the east coast of
North America (Margalef, 1961; Silva, 1967; Ho and Zubkoff, 1979;
Tomas and Smayda, 2008 reporting on a 1981 bloom). Since that
time, blooms along both of these continents have expanded. For
example, during the 1990s, annual, fish-killing blooms of C.
polykrikoides became persistent features within western Japanese
and South Korean coastal waters (Kim, 1997; Lee, 2008; Matsuoka
et al., 2010; Onitsuka et al., 2010). Oceanographic studies have
concluded that the Tsushima Warm Current is capable of
transporting populations from Japan to Korea (Matsuoka et al.,
2010; Onitsuka et al., 2010), a finding corroborated by Nagai et al.
(2009) who used microsatellites to demonstrate that these two
populations are genetically similar and are likely to undergo
frequent exchanges of genetic materials. This core Japanese–
Korean population appears to have expanded east of this region
into Chinese coastal waters within the East China Sea (Qi et al.,
1993) and later into the coastal South China Sea region (Huang and
Dong, 2000; Ou et al., 2010). Cochlodinium blooms also began
occurring on a regular basis during the end of the last decade south
of Japan and Korea in the Philippines (Azanza et al., 2008) and
Malaysia (Anton et al., 2008). Although blooms of Cochlodinium
have not been noted north of Japan, several studies have recovered
cysts of C. polykrikoides in Russian coastal waters including the Sea
of Japan (Orlova et al., 2004, 2009). On the Australia continent,
Hallegraeff (1992) described fish kills associated with blooms of
Cochlodinium cf. helix as an example of the expansion of new HABs
into this region.
C. polykrikoides was first described as a species in Puerto Rico
(Margalef, 1961), and since that time, blooms were noted in three
locations on the east coast of the US: NJ (Silva, 1967), RI (Tomas and
Smayda, 2008 reporting a bloom from 1981), and VA (Ho and
Zubkoff, 1979). Since these reports, there has been a significant
expansion of blooms reported along the US east coast often in wellmonitored regions where blooms had not been noted previously. On
eastern Long Island, this species had never been observed prior to
2004 despite robust HAB monitoring programs established in the
1980s. This species now forms annual, dense blooms in the Peconic
Estuary and Shinnecock Bay (2004–2011 Gobler et al., 2008; Tang
and Gobler, 2010; Gobler, pers. obs.) and has recently (2011)
expanded across Long Island into Great South Bay (Gobler, pers.
obs.). To the north of this region, blooms have been recently
observed in Point Judith Pond, RI (Hargraves and Maranda, 2002) as
well as in Narragansett Bay and Massachusetts coastal waters
(Gobler, pers. obs.), although scientific reports of the latter site have
yet to be published. To the south of this region, blooms of C.
polykrikoides had been sporadic events in the York River, MD (Ho and
Zubkoff, 1979) but have recently become regular, annual events
within multiple tributaries of Chesapeake Bay (Marshall et al., 2005;
Marshall, 2009; Mulholland et al., 2009). C. polykrikoides blooms
have also been newly reported in the southeast US specifically
within the Skidaway Estuary (GA, USA; Verity, 2010) and the Indian
River Lagoon (FL, USA; Phlips et al., 2011).
On the west coast of North America, Cochlodinium spp. cells had
been observed in southern California coastal waters during the
twentieth century (i.e. La Jolla Bay, California; Holmes et al., 1967;
Kofoid and Swezy, 1921). Since that time, the reported range of
Cochlodinium cells and blooms has expanded north into Canada and
77
south to Costa Rica. For example, Whyte et al. (2001) described
blooms that killed aquacultured fish near Vancouver Island (Canada)
while Curtiss et al. (2008) and Kudela et al. (2008) reported annual
Cochlodinum blooms in Monterey Bay (CA, USA), with recurrent
sightings throughout central and Southern California since 2004. In
Mexico, C. polykrikoides blooms have been reported within the Gulf
of California (Gárate-Lizárraga et al., 2004) as well as on the Pacific
coast, near Banderas Bay (Del Carmen Cortes Lara et al., 2004;
Gomez-Villarreal et al., 2008). Further south along the Pacific coast,
blooms have also been observed in the coastal waters of Costa Rica
(Vargas-Montero et al., 2004, 2006; Montero et al., 2008) and
Guatemala (Rosales-Loessener et al., 1996).
Beyond southeast Asia and North America, the most recent
decade has also seen several ‘first reports’ of Cochlodinium blooms
in the waters of southwestern Asia and Europe. For example, in the
Indian Ocean, Bhat and Matondkar (2004) noted the occurrence of
C. polykrikoides in the coastal waters of Goa, India, while Ramaiah
et al. (2005) reported blooms along the Malabar Coast of India.
Large blooms of C. polykrikoides have also been recently noted in
the Black Sea (Vershinin et al., 2004, 2005; Terenko, 2005),
Croatian coastal waters of the Adriatic Sea (Saracino and Rubino,
2006) and in the Mediterranean across the Gulf of Olbia (coastal
Italy; Sannio et al., 1997). One of the more significant HAB events of
the last decade occurred throughout the Arabian Gulf and Gulf of
Oman from 2008 through 2009 when a massive bloom of C.
polykrikoides caused fish kills, the death of coral reefs, and
disruption of desalination plants in Iran, Saudi Arabia, and the
United Arab Emirates (Richlen et al., 2010).
It is tempting to ascribe the expansion of Cochlodinum across
the globe during the past two decades to anthropogenic factors
such as enhanced nutrient loading (Heisler et al., 2008), ballast
water transport (Smayda, 2007), or climate change (Hallegraeff,
2010). While there may be some experimental and/or anecdotal
evidence to partly support each of these hypotheses in several
cases, the very recent occurrence of most of these events and
absence of research on the events beyond rudimentary description
of the blooms in most cases prohibits firm conclusions regarding
the causes of these blooms and their expansion from being drawn.
Given the striking visual attributes of Cochlodinum blooms (large,
dense, surface patches of red water), the occurrence of these events
in new regions is more likely to be a real phenomenon than a
function of improved methods of detection compared to other
HABs that form less distinguishable blooms.
5. Ecophysiology
A previous review of the ecophysiology of Cochlodinium (Kudela
et al., 2008) identified a scarcity of data on the physiological
response of Cochlodinium to such basic factors as temperature,
salinity, and light, with very little information about the nutritional
preferences or behavioral adaptations of this organism. In the past
few years the global expansion of C. polykrikoides blooms has
resulted in a substantial number of new studies. Based on the most
recent published data, it is now possible to provide temperature
and salinity ranges for naturally occurring blooms of the three
main ribotypes of C. polykrikoides and for C. fulvescens (Table 2).
5.1. Temperature and salinity
There is clear overlap in the temperature and salinity ranges for
the reported C. polykrikoides blooms. The data generally support
the description of this organism as eurythermal and euryhaline,
well adapted to warm (>208 C) moderate (30–33) salinities often
associated with offshore, possibly tropical or subtropical waters
(e.g. Kim et al., 2004; Kudela et al., 2008; Nagai et al., 2009).
However, some blooms, particularly in North America, have been
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R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
Table 2
Temperature and salinity ranges for naturally occurring Cochlodinium bloom events.
Location (year)
American/Malaysian ribotype
Phosphorescent Bay, Puerto Rico (1958)
Pettaquamscutt Cove, Rhode Island (1980–1981)
Manzanillo Bay, Mexico (1999–2000)
Gulf of California, Mexico (2000)
Peconic Estuary, New York (2002–2006)
Sabah, Malaysia (2005)
Sepanggar Bay, Sabah, Malaysia (2006)
Chesapeake Bay, USA (2007)
Kota Kinabalu, Sabah, Malaysia (2007)
Gulf of Oman (2008-2009)
Japanese–Korean ribotype
Yatsushiro Sea, Japan (1978)
Gamak Bay, Korea (1984–2006)
Quanshou Bay, China (1990)
South Coast, Korea (1995–1998)
South Coast, Korea (1995–2008)
Namhae Island, Korea (1996)
South Sea, Korea (1999)
Yatsushiro Sea, Japan (2000)
Inokushi Bay, Japan (2002)
Yeoja Bay, Korea (2003)
East Sea/Sea of Japan, Korea (2003)
South Sea, Korea (2004)
Usuka Bay, Japan (2005)
Philippines ribotype
Palawan, Philippines (2005)
C. fulvescens
Vancouver Island, Canada (1999)
Monterey Bay, California (2004, 2006)
Temperature (8C)
Salinity
Source
29.8–30.1
3.0–28.0
25.5
29.0–31.0
19.0–29.1
30.2–31.2
24.27–32.75
27.0–29.5
30.2 (1.1)c
27.0
35.3–35.8
25.0–31.0
34.5–34.7
Margalef (1961)a
Tomas and Smayda (2008)b
Morales-Blake et al. (2001)a
Gárate-Lizárraga et al. (2000)a
Gobler et al. (2008)
Anton et al. (2008)
Rashed-Un-Nabi et al. (2010)
Mulholland et al. (2009)
Adam et al. (2011)
Richlen et al. (2010)
23.0–29.9
20.2–27.0
22.4–26.7
22.5–27.0
23.1–27.1
24.0–25.2
22.8–26.5
24.5–26.6
18.9–20.3
24.0–26.0
12.0–21.6
18.10–19.99
12.3–27.6
31.7–34.2
16.34–35.53
31.3–33.8
30.0–33.4
28.0–34.13
28.94–31.78
32.0–33.0
32.6–34.6
27.8–30.3
30.2-32.8
32.81–33.66
32.52–34.40
Honda et al. (1980)a
M. Lee et al. (2009) and Y.S. Lee et al. (2009)
Du et al. (1993)a
NFRDI (1998) and Suh et al. (2000)a
Lee et al. (2010)
Park et al. (2001)
Lee et al. (2001)
Kim et al. (unpublished)a
Miyamura, pers. comm. to Kim et al. (2004)a
Kim et al. (2006)
Y.S. Kim et al. (2010)
Lee and Lee (2006)
Yamatogi et al. (2006), Table
31.0–36.0
33.0–36.0
Azanza et al. (2008)
11.4–13.5
14.0–18.0
29.6–31.4
32.8–33.6
Whyte et al. (2001)
Kudela et al. (2008)
23.7–26.5
32.4–32.8
22.55–34.26
18.9–27.9
29.8 (2.1)c
39.0
a
As reported in Kim et al. (2004).
Optimal ranges reported as 15–28 8C, 25–30 salinity.
c
Mean (SD) reported.
b
associated with the intrusion of cool water masses and more
brackish salinities. For example, within US east coast estuaries,
blooms occur over a broader and lower salinity range (19–30;
Gobler et al., 2008; Tomas and Smayda, 2008; Mulholland et al.,
2009; Morse et al., 2011) than most other locations (Table 2). This
suggests that, despite the genetic similarity the LSU gene of clones
within the American/Malaysian ribotype, there may be ecological
differences among these populations. Regarding temperature, in
Mexico, blooms were associated with anomalously cool (decrease
from 25.5 to 21.2 8C) waters relative to conditions prior to the
bloom event (Morales-Blake et al., 2001), while Tomas and Smayda
(2008) reported motile cells in waters as cold as 3.0 8C, and
Yamatogi et al. (2006) reported persistent populations in winter
(>12.58 C) from western Japan. Kim et al. (2004) suggested that
vegetative cells in Korean waters could overwinter at temperatures
>14–15 8C, and reported that morphological changes (including
shortening of chain length) were a stress response to low
temperature and salinity. Tomas and Smayda (2008) similarly
reported variable chain length correlated to temperature, with
single cells found predominantly in suboptimal conditions. Thus
based on field observations, C. polykrikoides exhibits a wide range
of temperatures over which it can survive in a vegetative state,
despite a relatively narrow ‘‘optimal’’ temperature and salinity
preference. Distinct from C. polykrikoides, C. fulvescens appears to
be associated with cool coastal upwelling conditions (Whyte et al.,
2001; Kudela et al., 2008; Trainer et al., 2010) although there is
very little field data for this organism and no reported laboratory
data for growth response to temperature or salinity.
Laboratory studies of Cochlodinium ecophysiology have largely
been restricted to the Japanese–Korean ribotype. Kim et al. (2001b)
and Lee et al. (2001) reported results similar to field observations:
maximal growth rates were 0.30–0.55 d 1, with wide tolerance
exhibited for both salinity (15–50, with an optimum between 25
and 40), and growth temperature (ca. 10–30 8C, optimal at 25 8C).
These results were corroborated by data from Kim et al. (2004),
who examined the temperature and salinity tolerance for a
Japanese strain of C. polykrikoides in culture. Optimal conditions
were again identified as 25 8C and a salinity of 34, which resulted in
maximal growth (0.41 d 1). Yamatogi et al. (Yamatogi and Maruta,
2002; Yamatogi et al., 2005, 2006) have similarly documented the
growth characteristics of multiple C. polykrikoides strains isolated
from Imari Bay, Usuka Bay, and Isahaya Bay Japan. The growth
response to temperature and salinity was similar, with a
temperature optimum of 27.5 8C for all strains and salinity optima
of 32–34.4. Maximal growth was considerably higher than
previously reported, however, ranging from 0.56 d 1 (Isahaya
Bay) to 0.90 d 1 (Imari Bay).
Based on these observations from the field and laboratory, C.
polykrikoides exhibits a reasonably high temperature and salinity
optimum (>258 C, 25–40). Cochlodinium is capable of surviving and
initiating new bloom events at much lower temperatures, albeit
with some evidence of stress (e.g. shorter chain length), but has not
been reported to grow below 10 8C (Kim et al., 2004; Yamatogi et al.,
2006; Tomas and Smayda, 2008). Growth rates are comparable to
other HAB dinoflagellates (Smayda, 1997), but the limited laboratory
data suggest there is a fairly broad (factor of 2) range for maximal
growth. Although comparisons by ribotype are limited to field
studies, there does not appear to be much differentiation in
ecophysiological space between ribotypes for C. polykrikoides, other
than the lower temperature and fresher salinities observed for
blooms on the US east coast; C. fulvescens does appear to be adapted
to lower temperatures than C. polykrikoides based on the extremely
limited data available (Table 2).
5.2. Irradiance
As with temperature and salinity, there are few studies
explicitly examining the effect of irradiance on Cochlodinium
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
growth. Several authors reported no sign of photoinhibition at
moderate irradiance levels (Kim et al., 2001b, 2004; Lee et al.,
2001; Oh et al., 2006; Kudela et al., 2008), a characteristic
consistent with the propensity of this species to form surface water
blooms. Half-saturation values (Ek) are consistent across studies,
ranging from 29.2 to 45 mmol photons m 2 s 1 (Kim et al., 2004;
Yamatogi et al., 2005; Oh et al., 2006) for C. polykrikoides. Oh et al.
(2006) reported optimal growth under blue (as opposed to yellow
or red) light, which the authors attributed to adaptation for open
ocean rather than coastal waters. Y.S. Lee et al. (2009) similarly
reported C. polykrikoides bloom conditions to be associated with
high insolation and warm, euryhaline conditions in Gamak Bay,
Korea. Park et al. (2001) correlated increasing chain length with
increased cell division rates, and suggested that shorter chains,
with higher area:volume ratios (per cell), could be an adaptive
strategy to maximize photosynthesis in surface waters. Cochlodinium thus appears to be well adapted to a high-light environment,
and does not exhibit photoinhibition at typical near-surface
irradiances.
5.3. Nutrient utilization
General aspects of Cochlodinium nutrient utilization have been
summarized by Kudela et al. (2008, 2010). As with other
ecophysiological characteristics, nutrient acquisition by Cochlodinium has been most extensively examined for the Japanese–
Korean ribotype. Lee et al. (2001) obtained similar growth
responses for nitrate and ammonium, with no difference in
maximal growth rates. Kim et al. (2001b) reported half-saturation
constant (Ks) values for growth of 2.10, 1.03, and 0.57 mM for
nitrate, ammonium, and phosphate, respectively, with a preference for ammonium versus nitrate, and concluded that, while the
relatively low Ks values suggest it is adapted to neritic waters
Cochlodinium is not a true ‘‘eutrophic’’ dinoflagellate. This is
supported by previous reviews (Kudela et al., 2008, 2010) which
noted that both C. polykrikoides and C. fulvescens are capable of
utilizing a variety of N compounds, but exhibit characteristics
more indicative of pelagic rather than neritic/eutrophic dinoflagellates. Mulholland et al. (2009) also found that C. polykrikoides
likely utilizes a variety of nutrients in the Chesapeake Bay,
including dissolved organic N, P, and C, to meet its nutritional
requirements, while Gobler et al. (submitted for publication) found
that bloom populations displayed the greatest N uptake rates for
the compounds in the greatest abundance (nitrate, ammonium,
urea, or glutamic acid).
In contrast to the view of Cochlodinium as more pelagic than
neritic, several recent studies have linked Cochlodinium blooms to
eutrophication or demonstrated that Cochlodinium readily accesses
nutrients typically associated with neritic or eutrophied environments. For example, both Imai et al. (2006) and Verity (2010)
reported an increase in C. polykrikoides as part of a long-term trend
of increasing HAB organisms in the Seto Inland Sea and a Georgia
(USA) estuary, respectively. Imai et al. (2006) further noted that the
low N and P requirements to reach a ‘‘significant’’ bloom event
(consistent with the pelagic nutrient strategy identified for the
genus) could result in coastal blooms of C. polykrikoides with only
minor nutrient enrichment.
B.-K. Lee et al. (2008) demonstrated that a Japanese–Korean
strain of C. polykrikoides grew well on multiple sources of organic
nitrogen and phosphorus, as well as with various forms of selenium,
but needed to be gradually acclimated to increasing ammonium
concentrations (up to 20 mM) to achieve optimal growth. Interestingly, growth on ammonium (as high as 80 mM) was significantly
enhanced by the presence of moderate (20 mM) concentrations of
nitrate. A North American isolate of C. polykrikoides was shown to
grow faster on glumatic acid (0.5 d 1) as a sole N source than
79
nitrate, ammonium, and urea all of which yielded similar growth
rates (0.4 d 1; (Gobler et al., submitted for publication). In the
southern sea of Korea, Lee et al. (2010) linked groundwater discharge
to the maintenance of red tides. Although the groundwater itself was
dominated by dissolved inorganic nitrogen and phosphorus (DIN
and DIP), the dominant nutrients available to the red tide organisms
were organic substrates, which the authors suggested favoured
dinoflagellates over diatoms. This selection for Cochlodinium versus
diatoms under low DIN and DIP conditions was also proposed by Kim
et al. (2006) for South Korean waters. Several authors (e.g. Viques
and Hargraves, 1995; Lee and Lee, 2006; Anton et al., 2008; M. Lee
et al., 2009; Mulholland et al., 2009; Morse et al., 2011) have also
associated increases in Cochlodinium with rainfall, attributed to
changes in both the physical environment and the influx of
discharge-associated nutrients. Consistent with these observations,
experimental enrichment with N concentrations has been shown to
significantly enhance net growth rates and photosynthesis of
Cochlodinium during blooms in NY coastal waters (Gobler et al.,
submitted for publication).
A recent review (Hansen, 2011) places Cochlodinium, along with
several other red tide organisms, in a group (‘‘Type 1’’) that generally
utilizes mixotrophy to moderately enhance growth at low irradiance
levels, while several reviews (Stoecker et al., 2006; Burkholder et al.,
2008) suggest phagotrophy and osmotrophy are common in HAB
organisms, particularly for those forming red tides. Research on the
Japanese–Korean ribotypes of C. polykrikoides has demonstrated
mixotrophic or phagotrophic feeding on multiple types of planktonic prey up to 11 mm in diameter (including cryptophytes,
Synechococcus sp., a diatom, a dinoflagellate, a raphidophyte, and a
prymnesiophyte); however, C. polykrikoides was unable to consume
larger (>12 mm) dinoflagellates (Larsen and Sournia, 1991; Jeong
et al., 2004, 2005; Yoo et al., 2009). Careful microscopy demonstrated C. polykrikoides specifically ingests cells through its sulcus (Jeong
et al., 2004). C. polykrikoides has also been shown to consume
bacteria during bloom events in Korean waters, although its rate of
consumption provided <2% its daily carbon requirements, a finding
that left the investigators to conclude C. polykrikoides was not a
significant bacterivore perhaps because marine bacteria are at the
lower range of its ideal prey size (Seong et al., 2006). Mulholland
et al. (2009) also speculated that organic carbon (from either
mixotrophy or osmotrophy) was likely subsidizing growth during
bloom conditions in the Chesapeake Bay.
While studies in Korea have clearly demonstrated the ability of
C. polykrikoides to phagotrophically consume planktonic prey,
many unanswered question persist regarding the ecological role of
mixotrophy within the Cochlodinium genus. Further studies are
needed to explore phagotrophy in Cochlodinium beyond Korea and
to better understand the extent to which Cochlodinium grazes
various planktonic prey during blooms, the relative contribution of
phagotrophy, osmotrophy, and inorganic nutrient acquisition, and
the circumstance under which the various ribotypes switch
between strategies during bloom events.
In summary, Cochlodinium exhibits wide flexibility in its
nutrient acquisition strategies. While its nutrient kinetics are
generally consistent with pelagic origin, this genus readily utilizes
organic nitrogen and phosphorus compounds, and appears to do
well in moderately eutrophied environments, particularly when
DIN and DIP are low. While its ability to phagotrophically consume
prey further extends its nutritional flexibility, the relative
importance of all of these nutrient acquisition strategies during
bloom events has not been well characterized.
5.4. Vertical migration
Chain-forming dinoflagellates have an adaptive advantage in
terms of swimming speed and ability to withstand vertical
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R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
velocities (Fraga et al., 1989; Anderson et al., 2005). Both C.
polykrikoides and C. fulvescens form chains of 2 or more cells,
generally decreasing chain length with sub-optimal growth
conditions (e.g. Whyte et al., 2001; Tomas and Smayda, 2008;
Jiang et al., 2010b), and increasing chain length during periods of
active vertical migration (Park et al., 2001). This vertical migratory
behavior increases the swimming speed of cells as eight cell chains
swim at twice the rate of solitary cells (Sohn et al., 2011). This
swimming may provide some photosynthetic advantages (Park
et al., 2001; Y.S. Kim et al., 2010), as well as access to the deep
nutricline or within sediments at night (e.g. Eppley and HolmHansen, 1968; Cullen and Horrigan, 1981; Smayda, 1997; Kudela
et al., 2008). Jiang et al. (2010b) further demonstrated that chain
length varied in response to grazing pressure, attributing increased
chain length to an adaptive strategy to minimize grazing (size
mismatch), or to selective grazing pressure on single cells. Those
authors also reported that chain length was positively correlated
with availability of B-vitamins (B1, B7, and B12), again linking chain
length to overall physiological status. Thus Cochlodinium appears
to employ chain formation as an adaptive strategy to multiple
environmental conditions including optimization of light and
nutrients, minimization of grazing and possibly shear-stress
(Sullivan et al., 2003), and as a general response to population
growth rates.
6. Life history
The life history of Cochlodinium has been a subject of recent
study, but is still not well characterized. The recurrence of annual
blooms following initial colonization of an area by Cochlodinium
strongly implicates seed populations in the establishment of this
organism in new habitats. The life cycle has been proposed to
include two different vegetative stages, a temporary (hyaline) cyst,
and as many as three different resting cyst morphotypes (RosalesLoessener et al., 1996; Matsuoka and Fukuyo, 2000, 2002; C.-H.
Kim et al., 2002; Kim et al., 2007). Kim et al. (2007) identified
multiple types of C. polykrikoides in the laboratory, including the
most commonly observed athecate vegetative cells, armored
vegetative cells (identified before and after blooms), temporary
(hyaline) cells, and cysts. Tomas and Smayda (2008) have also
reported hyaline cysts in field samples, while Park and Park (2010)
detected C. polykrikoides by PCR in sediment samples, and inferred
that resting stages or cysts were most likely present.
It should be noted that, despite the assumption of cysts or
resting stages as part of the life cycle of Cochlodinium, few
investigators have confirmed the importance of these life stages in
recurrent bloom regions and no data are currently available to
demonstrate unambiguously that the two most well-described
and notorious species, C. polykrikoides and C. fuvelscens, produce
resting cysts. Matsuoka et al. (2010) suggested that seed
populations in the East China Sea might be associated with
overwintering as vegetative cells at moderate (>10 8C) temperatures, and cast doubt on the identification of hyaline cysts or
resting stages reported by others for that region (c.f. Matsuoka
et al., 2010). Similarly, Nagai et al. (2009) inferred from
microsatellite analysis that local populations of C. polykrikoides
overwinter and survive as vegetative cells in local waters of Japan
and Korea. Seaborn and Marshall (2008), in contrast, claimed
identification of C. polykrikoides cysts from sediments in east
coast, USA estuaries, while Rubino et al. (2010) also reported a cyst
from the Mediterranean tentatively identified as C. polykrikoides,
which was successfully germinated to a gymnoid vegetative cell.
Neither identification, however, was confirmed with detailed
morphological and molecular examinations.
Cochlodinium is typically described as a pelagic organism that
can be transported shoreward and appear as a red tide (Park et al.,
2005; Vargas-Montero et al., 2006; Kim et al., 2007). Similar to
Korea, Cochlodinium has been observed, albeit infrequently, for
many years (e.g. Holmes et al., 1967) in coastal waters of California.
The recurrence of this organism in waters of central California is
reminiscent of the evolution from background level to multi-year
bloom events as exemplified in coastal waters of Korea. Similar
behavior has been observed in other ‘‘new’’ locations for
Cochlodinium blooms as well (see Section 4). The available evidence
suggests that Cochlodinium produces temporary and resting cysts
and this life history may be, in part, related to the recent expansion
of this genus globally. However, the production of resting cysts by
the two most noxious species, C. polykrikoides and C. fulvescens,
requires more convincing data, particularly reports of an armored
stage and the presence of cysts in sediments. Regional patterns are
also clearly related to pelagic seed banks. It is not clear at this time
what the relative importance of these proposed life stages play in
Cochlodinium dynamics, and there remain questions about the
specific morphologies (and molecular characterization) of the
multiple proposed life stages.
7. Food web interactions
7.1. Cochlodinium toxins
Cochlodinium has been implicated in kills of wild and
impounded fish around the globe (Onoue et al., 1985; Yuki and
Yoshimatsu, 1989; Guzmán et al., 1990; Qi et al., 1993; Whyte
et al., 2001; Gárate-Lizárraga et al., 2004) and has been the cause of
fisheries losses exceeding US$100 million annually in Korea (Kim,
1998; Kim et al., 1999). Studies have also demonstrated that this
alga causes rapid mortality in reef building coral (Bauman et al.,
2010; Richlen et al., 2010), copepods (Jiang et al., 2009, 2010a),
other phytoplankton (Tang and Gobler, 2010), shellfish larvae (Ho
and Zubkoff, 1979; Tang and Gobler, 2010), bivalves (Ho and
Zubkoff, 1979; Tang and Gobler, 2010), and cultured fish (Gobler
et al., 2008; Dorantes-Aranda et al., 2009; Tang and Gobler, 2009a).
Despite the severe and broad nature of the toxicity of Cochlodinium,
the precise compounds responsible for this toxicity have yet to be
confirmed.
Landsberg (2002) categorized Cochlodinium species as algae
with multiple toxins. Two early studies reported three toxic
fractions (i.e. neurotoxic, hemolytic, and hemagglutinative; Onoue
and Nozawa, 1989a) and zinc-bound paralytic shellfish poisoning
(PSP) toxins (Onoue and Nozawa, 1989b) from the red tide waters
of Cochlodinium type’ 78 Yatsushiro, which was considered to be
conspecific with C. polykrikoides (Matsuoka et al., 2008). However,
the PSP toxins and two of the non-PSP toxic fractions have not been
identified further or described since their initial report (Onoue and
Nozawa, 1989a,b), while the third fraction, hemolytic agents, has
been documented to be associated with fatty acids (Lee, 1996).
Because these toxic fractions were originally extracted from a
mixed biomass sample concentrated from a large volume of field
bloom water (1000 L; Onoue and Nozawa, 1989b), it is possible
that these toxins or toxic fractions came from other sources. Other
studies have indicated that reactive oxygen species (ROS; i.e.
superoxide anions, hydrogen peroxide, and many other compounds) are produced by C. polykrikoides cells and may be one of
the factors responsible for lipid peroxidation and subsequent fish
kills caused by this species (Kim et al., 1999). Interestingly, C.-H.
Kim et al. (2002) and D. Kim et al. (2002) found that O2 and H2O2
production by C. polykrikoides was much lower than that by
Chattonella marina, a species well known for ROS production.
Importantly, however, the D. Kim et al. (2002) study was
performed using a culture of C. polykrikoides with a cell density
more than 8000 cells ml 1 for the detection of ROS, which was
likely in near-stationary growth phase and thus may not have been
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
actively generating ROS, as seen in Kim et al. (1999). Given the
gradual accumulation of polysaccharides in C. polykrikoides culture
medium (D. Kim et al., 2002) and cells (Gobler et al., 2008), it has
been suggested that there may be multiple, biologically active
metabolites secreted by C. polykrikoides such as cytotoxic agents
and mucus substances such as polysaccharides that are responsible for fish kills caused by this species (D. Kim et al., 2002). This is
consistent with the observations that the polysaccharides produced by some dinoflagellates can induce apoptosis in human
lymphoid cells (Sogawa et al., 1998a,b). However, Cochlodinium
cultures are most toxic in the earliest stage of growth (Kim et al.,
1999; Tang and Gobler, 2009a,b) whereas polysaccharides tend to
accumulate as cultures age (Mague et al., 1980). Moreover,
Cochlodinium cultures can be toxic to fish across a fine (5 mm)
mesh on a short time scale (hours; Tang and Gobler, 2009b) and
mucus-like aggregates would be unlikely to quickly and passively
penetrate a fine mesh barrier, collectively suggesting polysaccharides may not be the primary toxic agent in C. polykrikoides.
While falling short of positively identifying the precise toxins
produced by Cochlodinium, recent research on North American
isolates and blooms of this species have shed further light on its
mode of toxicity. Histopathological examination of fish gills
exposed to Cochlodinium for <1 h have revealed acute degeneration of epithelial and chloride cells along with complete loss of the
structure and function (i.e. respiration, nitrogen excretion, ion
balance) of gill lamellae (Gobler et al., 2008; Tang and Gobler,
2009b). Such rapid degradation of the gill lamellae is characteristic
of an external insult to the gills such as exposure to a parasite
(Roberts, 2001) and is similar to the characteristics of fish gills
exposed to lethal levels of ROS (Tort et al., 2002). Shellfish exposed
to Cochlodinium for several days have displayed hyperplasia,
haemorrhaging, squamation, and apoptosis in both gill and
digestive tissues (Gobler et al., 2008), indicating that Cochlodinium
toxins target a broad range of organisms and tissue types.
Prior reports documented the production of hemolytic toxins by
Cochlodinium that cause lesions and alterations in epithelial
barriers of vertebrates and invertebrates (Landsberg, 2002;
Dorantes-Aranda et al., 2009) similar to those observed in gills
and digestive epithelia of fish and shellfish exposed to this alga
(Gobler et al., 2008; Tang and Gobler, 2009b). The ichthyotoxicity
of C. polykrikoides isolates from North America has been shown to
be highly labile (persisting for <1 h in filtered culture water),
mitigated by ROS-scavenging enzymes (e.g. peroxidase and
catalase), proportional of cellular growth rates, dependent on cell
viability (i.e. dead cells are non-toxic), not dependent of physical
contact, and can be greatly reduced by the presence of nonCochlodinium phytoplankton (Tang and Gobler, 2009a,b), and most
of these characteristics were observed with regard to the toxicity of
the same isolate to copepod (Jiang et al., 2009). Collectively, these
observations suggest that Cochlodnium toxins are extracellular,
highly reactive, and labile compounds that are produced continuously by actively growing C. polykrikoides cultures, similar to, but
not necessarily being, ROS.
Some harmful algae display a wide range of toxicity or noxious
effects among culture strains and field populations (Burkholder
and Glibert, 2009) and different species of Cochlodinium and even
‘ribotypes’ of C. polykrikoides (Matsuoka et al., 2008) may differ in
both their relative toxicity and their mode of toxicity. Indeed,
preliminary experiments comparing strains of Cochlodinium
isolated from the Atlantic and Pacific as well as North America
and Asia have shown strong differences in toxicity, from highly
lethal to non-toxic among strains of different origins (Tang and
Gobler, unpublished data). Clearly, much work is still needed to
resolve the precise mechanism of toxicity in Cochlodinium and to
explore the extent to which conclusions regarding individual
strains may be applicable to other populations. Regardless, given
81
that the smallest individuals and larvae of fish and shellfish as well
as plankton are most vulnerable to the toxicity of Cochlodinium,
the ecosystem-level impacts of these blooms almost certainly
extend well-beyond obvious fish kills observed during bloom
events.
7.2. Grazing on Cochlodinium
Although many HABs are thought to be caused by factors that
enhance cellular growth rates such as nutrients, light, and
temperature, an absence of strong top down control by grazing
populations may also contribute towards bloom formation (Gobler
et al., 2002; Sunda et al., 2006; Smayda, 2008). There are several
lines of evidence to indicate that Cochlodinium blooms are
promoted and sustained by weak grazing control. A series of
studies by Jiang et al. (2009, 2010a,b) demonstrated that C.
polykrikoides cultures isolated from North America can cause
depressed survival, grazing rates, egg production, and egg hatching
in the copepod Acartia tonsa and specifically demonstrated that C.
polykrikoides increases its chain length in the presence of this
species, an adaptation that further depresses grazing. These
findings are consistent with a parallel study on Korean isolates
that depressed egg production and viability in A. omorii (Shin et al.,
2003). Although the mixotrophic dinoflagellate Fragilidium cf.
mexicanum has been shown to graze on multiple toxic dinoflagellates including Lingulodinium polyedrum, Akashiwo sanguinea,
Prorocentrum micans, P. minimum, and Scrippsiella trochoidea, this
species was incapable of grazing C. polykrikoides (Jeong et al.,
1999). Bivalve larvae are known to be ephemerally significant
planktonic grazers (Carriker et al., 2001), but studies in North
America have demonstrated that bloom densities of C. polykrikoides cause rapid and near complete mortality in the larvae of
multiple species of North America bivalves (Ho and Zubkoff, 1979;
Tang and Gobler, 2009a). The adult stages of bivalves may also
exert significant grazing pressure on phytoplankton (Officer et al.,
1982), particularly in the shallow estuaries where C. polykrioides
blooms occur (Smayda, 2008). However, the ability of C.
polykrioides blooms and isolates to cause mortality in multiple
species of filter feeding bivalves (Gobler et al., 2008; Tang and
Gobler, 2009b) indicates these grazers are also not likely to exert
significant control over this species. While the large size of C.
polykrioides cells (40 mm) and chains (100s of mm) make it
vulnerable to grazing by planktivorous fish (Samson et al., 2008),
its ability to cause rapid mortality in these animals (Gobler et al.,
2008; Dorantes-Aranda et al., 2009; Tang and Gobler, 2009b)
prohibits this grazing control as well.
Despite the diverse array of grazing organisms that are killed by
elevated densities of C. polykrioides, there are two planktonic
organisms that can graze this alga: a large ciliate isolated from
Korean coastal waters (Strombidinopsis jeokjo; Jeong et al., 2008)
and larvae from the mussel Mytilus galloprovincialis (Jeong et al.,
2004). With the exception of these two Korean plankton, the ability
of Cochlodinium blooms to suppress survival and grazing by
copepods (Shin et al., 2003; Jiang et al., 2009, 2010a,b), protozoan
zooplankton (Jeong et al., 1999), bivalve larvae (Ho and Zubkoff,
1979; Tang and Gobler, 2009a), adult bivalves (Gobler et al., 2008;
Tang and Gobler, 2009b), and fish (Gobler et al., 2008; DorantesAranda et al., 2009; Tang and Gobler, 2009b) indicates that C.
polykrioides blooms are partly promoted by an absence of benthic
and pelagic grazing control. Since the ability of C. polykrioides to
inhibit many of these grazers is density dependent, with maximal
suppression occurring at densities exceeding 103 cells ml 1
(Gobler et al., 2008; Jiang et al., 2009; Tang and Gobler,
2009a,b; Jiang et al., 2010a,b), it is likely that grazing suppression
contributes to bloom maintenance but not necessarily bloom
initiation.
82
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
7.3. Allelopathy of Cochlodinium
In addition to suppressing grazers, C. polykrikoides blooms and
strains isolated from North America have exhibited strong
allelopathic effects on natural communities and ten species of
cultured phytoplankton, causing complete lysis of competing algae
in 24 h at C. polykrikoides cell densities exceeding 103 cells ml 1
(Tang and Gobler, 2010). These results are consistent with the
observations of Yamasaki et al. (2007) who reported growth
inhibition and formation of abnormal cells in Akashiwo sanguinea
co-cultured with C. polykrikoides isolated from Korean coastal
waters but explained with a mechanism necessitating direct
physical cell contact. The allelopathic effects of C. polykrikoides on
target microalgae have been shown to be dependent on the relative
and absolute cell abundance of C. polykrikoides and target algal
species as well as exposure time (Tang and Gobler, 2010). Since C.
polykrikoides is generally slow-growing (m = 0.4 d 1; Kim et al.,
2001a; Lee et al., 2001; Gobler et al., submitted for publication; see
also Section 5) compared to other diatoms and dinoflagellates
(Smayda, 1997), it must rely on other mechanisms, such as
allelopathy to form mono-specific blooms. Since C. polykrikoides
causes dramatic mortality in planktonic grazers (as discussed
above), allelopathic effects on competitors together with grazing
deterrence could promote C. polykrikoides blooms through positive
feedback (Sunda et al., 2006) whereby higher C. polykrikoides cell
densities yield fewer competitors and predators which in turn
facilitates higher cell densities. Since the allelopathic effects of C.
polykrikoides on co-occurring algae depend on absolute and
relative cell abundances, allelopathy is more likely to contribute
towards bloom maintenance when cell densities are high, than
bloom initiation when cell densities are low. The ability of C.
polykrikoides to form localized areas of high cell densities by
diurnal migration (Gobler et al., 2008; Kudela et al., 2008),
however, could facilitate a scenario whereby C. polykrikoides could
have allelopathic effects on competing algae strong enough to
enhance bloom initiation.
8. Prevention, control, and mitigation
Given the highly destructive nature of ichthyotoxic Cochlodinium blooms, there has been significant interest in, and progress
towards, developing techniques for the prevention, mitigation, and
control (PCM) of these events. In Korea, the spraying of fluidized
clays has been employed by aquaculture fish farmers to prevent
the rapid mortality of valuable, caged fish (Y.-J. Lee et al., 2008). Y.J. Lee et al. (2008) specifically reported on rapid removal of nearly
all (95%) Cochlodinium cells during blooms using a mixture of
sophorolipid and yellow clay. Subsequent work by Song et al.
(2010) demonstrated that dredged sediments of mixed compositions (slaked lime, quicklime, aluminium sludge bentonite, and
zeolite) effectively removed lower C. polykrikoides cell densities
within an experimental setting but less effectively removed higher
cell densities. Beyond density-dependent removal rates, a second
concern with regard to clay applications is the ability of C.
polykrikoides to reform blooms once removed by clay. For example,
Y.S. Lee et al. (2009) demonstrated that a fraction of C. polykrikoides
cells survive precipitation and sinking caused by yellow loess clays
and may reform substantial cell densities (>2500 cells ml 1)
within weeks of removal. Studies exploring the effectiveness of
clay as a mitigation strategy against Cochlodinium blooms have not
been explored beyond Korea.
In Asia, macroalgae are often grown in large stands (10s of km2)
in coastal waters (Neori et al., 2004; Yang et al., 2006). In some
cases, macroalgae are deployed as a nutrient mitigation strategy
(Neori et al., 2004; Yang et al., 2006; Zhou et al., 2006) whereas in
other scenarios, macroalgae are part of integrated multi-trophic
aquaculture systems (IMTA) whereby macroalgae are cultivated
with aquacultured fish and/or shellfish with the macroalgae often
being harvested for commercial use (Kraemer et al., 2004; Neori
et al., 2004; Abreu et al., 2009) or as feed for aquacultured animals
(Robertson-Andersson et al., 2008; Bolton et al., 2009). Beyond
these benefits, many studies have identified the ability of
macroalgae or macroalgal extracts to inhibit the growth of HABs,
including C. polykrikoides. Macroalgae capable to restricting the
growth of this HAB include Corallina pilulifera (Jeong et al., 2000;
Oh et al., 2010), Ecklonia kurome (Nagayama et al., 2003), Ishige
foliacea and Endarachne binghamiae (Jeong et al., 2000), Ulva
fasciata (Alamsjah et al., 2008), U. pertusa (Jeong et al., 2000;
Alamsjah et al., 2008), and U. lactuca (Tang and Gobler, 2011). In
several cases, the compounds specific for the algicidal effects,
including polyunsaturated fatty acids (from C. pilulifera, U. fasciata
and U. pertusa (Alamsjah et al., 2008; Oh et al., 2010), and
phlorotannins (from E. kurome; Nagayama et al., 2003), have been
shown to be the responsible for the inhibitory effects.
Multiple species of algicidal bacteria as well as bacterially
derived compounds have been proven capable of experimentally
reducing densities of C. polykrikoides. In some cases, live bacterial
cultures originally isolated from Korean coastal waters including
Alteromonas sp. A14 and Micrococcus luteus SY-13 have been shown
eliminate up to 90% of C. polykrikoides cells in culture (M.J. Kim
et al., 2008; B.-K. Lee et al., 2008). While the release of live bacteria
in an ecosystem setting may not be an ideal PCM approach due to
complex and unknown effects within the microbial food web,
several studies have advanced the research of C. polykrikoideskilling bacteria by identifying bacterially produced compounds
that have lethal effects on this alga. For example, the bioactive,
alkaloid compound, prodigiosin, has been isolated from multiple
species of marine bacteria originating from Korean coastal waters
including the gamma-Proteobacteria, Hahella chejuensis KCTC 2396
and this compound is lethal to C. polykrikoides at concentrations of
1 mg L 1 (D. Kim et al., 2008; S. Kim et al., 2008). L-2Azetidinecarboxylic acid and bacillamide have been isolated from
the bacteria Polygonatum odoratum var. pluriflorum and Bacillus sp.
SY-1, respectively, and are capable of lysing C. polykrikoides cells at
slightly higher concentrations (10 mg L 1; Jeong et al., 2003; Kim
et al., 2006). Importantly, however, not all algicidal bacteria are
active against C. polykrikoides. Imai and Kimura (2008) reported
that six isolates of bacteria that control many species of HABforming dinoflagellates were ineffective against C. polykrikoides.
Finally, several synthetic chemicals that actively lyse C.
polykrikoides have been identified including sophorolipids, cocamidopropyl betaine (Sun et al., 2004a,b), and the synthetic
algicidal drug TD53 (Han et al., 2010). Jeong et al. (2002) noted
that several HAB-forming dinoflagellates were more sensitive to
sodium hypochlorite produced by the electrolysis of seawater than
most other marine plankton and thus suggested this process may
also effectively mitigate C. polykrikoides blooms within smaller,
enclosed settings.
9. Open questions and conclusions
Cochlodinium appears to be well adapted to a wide variety of
ecophysiological conditions. Although its preferred niche is
primarily eurythermal and euryhaline, it can apparently overwinter at very low temperatures and there is evidence for distinct
ribotype or species-level adaptations to cooler and fresher
conditions in some habitats (e.g. the Japanese–Korean versus
the American–Malaysian ribotypes, and C. polykrikoides vs. C.
fulvescens). It is a strong vertical migrator capable of mixotrophy
and is flexible in its strategies for acquiring nutrients, readily
utilizing multiple forms of inorganic and organic substrates. It is
associated with both moderate, indirect nutrient loading and
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
eutrophied coastal waters. There is little evidence for photoinhibition of this species, which in combination with the strong
phototactic response and persistence in near surface waters
implicates high-light adaptation and the likely important role of
phototrophy of these populations.
Despite numerous studies carried out on multiple geographical
strains and ribotypes in the past few years, many questions about
this genus remain. First, this polyphyletic genus requires a refined
characterization both morphologically and genetically. While
there is good evidence for at least 3 ribotypes within C.
polykrikoides, a fourth has been proposed but not well characterized, and there is clear potential for multiple ribotypes in C.
fulvescens. Second, despite the new studies, the ecophysiology of
the Philippines ribotype (and to a lesser extent the other ribotypes)
has been poorly characterized. Third, the putative life stages of
Cochlodinium and how these relate to the apparent global
expansion of this genus require resolution. While the expansion
and persistence of established populations is consistent with the
formation of durable resting stages that could be transported by
both oceanic currents and ballast water, it is not possible to rule
out other mechanisms such as the persistence of difficult to detect
background pelagic populations. In a manner similar to other fishkilling algae, the precise mechanism(s) of toxicity exhibited by
Cochlodinium are still largely unknown, but its toxic effects likely
permit blooms to develop with minimal grazing losses and
competition. Finally, with regard to prevention, control and
mitigation, multiple promising approaches have been identified
but little is known regarding the long-term effectiveness of
physical controls such as clay flocculation, nor about the strainand ribotype-specific effectiveness of biological controls. To
summarize, Cochlodinium exhibits the ability to occupy new
ecological niches, allowing this formerly rare HAB genus to
proliferate in response to changes in the coastal environment.
Cochlodinium is well-positioned to take advantage of a wide range
of ecological conditions, and while much has been discovered
about this organism during the past two decades, many other
important questions regarding this alga need to be resolved.
Acknowledgements
We thank the members of the Kudela and Gobler labs for
feedback, specifically, Dr. Ying-Zhong Tang, Theresa K. HattenrathLehman, and Kendra Hayashi. Ms. Hayashi also prepared Figs. 3
and 4, while line drawings for Fig. 2 are provided by Corlis
Schneider. Partial funding was provided by NOAA MERHAB/
ECOHAB grants NA04NOS4780239 and NA-09N0S4780209NSF,
and NSF grant OCE-0421510 (RMK), as well as grants from the
Renaissance Charitable Foundation Incorporated and Suffolk
County, NY (CJG). This is MERHAB publication #155 and ECOHAB
publication #661.[SS]
References
Abreu, M.H., Varela, D.A., Henrı́quez, L., Villarroel, A., Yarish, C., Sousa-Pinto, I.,
Buschmann, A.H., 2009. Traditional vs. integrated multi-trophic aquaculture of
Gracilaria chilensis C. J. Bird, J. McLachlan & E. C. Oliveira: productivity and
physiological performance. Aquaculture 293, 211–220.
Adam, A., Mohammad-Noor, N., Anton, A., Saleh, E., Saad, S., Raehanah, S., Shaleh, M.,
2011. Temporal and spatial distribution of harmful algal bloom (HAB) species in
coastal waters of Kota Kinabalu, Sabah, Malaysia. Harmful Algae 10, 495–502.
Ahn, Y.-H., Shanmugam, P., Chang, K.-I., Moon, J.-E., Ryu, J.-H., 2005. Spatial and
temporal aspects of phytoplankton blooms in complex ecosystems off the
Korean coast from satellite ocean color observations. Ocean Sci. J. 40, 67–78.
Ahn, Y.-H., Shanmugam, P., Ryu, J.-H., Jeong, J.-C., 2006. Satellite detection of
harmful algal bloom occurrences in Korean waters. Harmful Algae 5, 213–231.
Alamsjah, M., Hirao, S., Ishibashi, F., Oda, T., Fujita, Y., 2008. Algicidal activity of
polyunsaturated fatty acids derived from Ulva fasciata and U. pertusa (Ulvaceae,
Chlorophyta) on phytoplankton. J. Appl. Phycol. 20, 713–720.
83
Anderson, D.M., 1989. Toxic algal blooms and red tides: a global perspective. In:
Okaichi, T., Anderson, D.M., Nemoto, T. (Eds.), Red Tides: Biology, Environmental Science, and Toxicology. Elsevier Science, New York, pp. 11–16.
Anderson, D.M., Glibert, P.M., Burkholder, J.M., 2002. Harmful algal blooms and
eutrophication: nutrient sources, composition, and consequences. Estuaries 25,
704–726.
Anderson, D.M., Pitcher, G.C., Estrada, M., 2005. The comparative systems approach
to HAB research. Oceanography 18, 148–157.
Anton, A., Teoh, P.L., Mohd-Shaleh, S.R., Mohammad-Noor, N., 2008. First occurrence
of Cochlodinium blooms in Sabah, Malaysia. Harmful Algae 7, 331–336.
Azanza, R.V., David, L.T., Borja, R.T., Baula, I.U., Fukuyo, Y., 2008. An extensive
Cochlodinium bloom along the western coast of Palawan, Philippines. Harmful
Algae 7, 324–330.
Babin, M., Cullen, J., Roesler, C., Donaghay, P., Doucette, G., Kahru, M., Mr, L., Scholin,
C., Sieracki, M., Sosik, H., 2005. New approaches and technologies for observing
harmful algal blooms. Oceanography 18, 210–227.
Bauman, A.G., Burt, J.A., Feary, D.A., Marquis, E., Usseglio, P., 2010. Tropical harmful
algal blooms: an emerging threat to coral reef communities? Mar. Pollut. Bull.
60, 2117–2122.
Bhat, S.R., Matondkar, S.B.P., 2004. Algal blooms in the seas around India –
networking for research and outreach. Curr. Sci. 87, 1079–1083.
Bok, T.-H., Paeng, D.-G., Kim, E., Na, J., Kang, D., 2010. Ultrasound backscattered
power from Cochlodinium polykrikoides, the main red tide species in the
Southern Sea of Korea. J. Plankton Res. 32, 503–514.
Bolton, J., Robertson-Andersson, D., Shuuluka, D., Kandjengo, L., 2009. Growing Ulva
(Chlorophyta) in integrated systems as a commercial crop for abalone feed in
South Africa: a SWOT analysis. J. Appl. Phycol. 21, 575–583.
Burkholder, J.A.M., Glibert, P.M., Skelton, H.M., 2008. Mixotrophy, a major mode
of nutrition for harmful algal species in eutrophic waters. Harmful Algae 8,
77–93.
Burkholder, J.M., Glibert, P.M., 2009. The importance of intraspecific variability in
harmful algae – preface to a collection of topical papers. Harmful Algae 8,
744–745.
Carriker, M.R., John, N.K., Michael, C., 2001. Functional morphology and behavior of
shelled veligers and early juveniles. Developments in Aquaculture and Fisheries Science, vol. 31. Elsevier, pp. 283–303 (Chapter 7).
Cho, E., Costas, E., 2004. Rapid monitoring for the potentially ichthyotoxic dinoflagellate Cochlodinium polykrikoides in Korean coastal waters using fluorescent
probe tools. J. Plankton Res. 26, 175–180.
Cho, E.S., 2002. Cluster analysis on the lectin binding patterns of marine microalgae.
J. Plankton Res. 25, 309–315.
Cho, E.S., Seo, G.M., Lee, S.G., Kim, H.G., Lee, S.J., Rhodes, L.L., Hong, Y.K., 1998.
Application of FITC-conjugated lectin probes for the recognition and differentiation of some Korean coastal red tide microalgae. J. Fish. Sci. Technol. 1,
250–254.
Costas, E., Rodas, V.L., 1994. Identification of marine dinoflagellates using fluorescent lectins. J. Phycol. 30, 987–990.
Cullen, J.J., Horrigan, S.G., 1981. Effects of nitrate on the diurnal vertical migration,
carbon to nitrogen ratio, and the photosynthetic capacity of the dinoflagellate
Gymnodinium splendens. Mar. Biol. 62, 81–89.
Curtiss, C.C., Langlois, G.W., Busse, L.B., Mazzillo, F., Silver, M.W., 2008. The
emergence of Cochlodinium along the California Coast (USA). Harmful Algae
7, 337–346.
Del Carmen Cortes Lara, M., Altamarino, R.C., Sierra-Beltran, A.P., 2004. Presence of
Cochlodinium catenatum (Gymnodiniales: Gymnodiniaceae) in red tides of
Bahia de Banderas, Mexican Pacific. Rev. Biol. Trop. 52 (Suppl. 1), 35–50.
Dierssen, H., Kudela, R., Ryan, J., Zimmerman, R., 2006. Red and black tides:
quantitative analysis of water-leaving radiance and perceived color for phytoplankton, colored dissolved organic matter, and suspended sediments. Limnol.
Oceanogr. 51, 2646–2659.
Dorantes-Aranda, J.J., Garcia-De La Parra, L.M., Alonso-Rodriguez, R., Morquecho, L.,
2009. Hemolytic activity and fatty acids composition in the ichthyotoxic
dinoflagellate Cochlodinium polykrikoides isolated from Bahia de La Paz, Gulf
of California. Mar. Pollut. Bull. 58, 1401–1405.
Du, Q., Huang, Y., Wang, X., 1993. Toxic dinoflagellate red tide by a Cochlodinium sp.
along the coast of Fujian, China. In: Smayda, T.J., Shimizu, Y. (Eds.), Toxic
Phytoplankton Blooms in the Sea. Elsevier, New York, pp. 235–238.
Eppley, R.W., Holm-Hansen, O., 1968. Some observations on the vertical migration
of dinoflagellates. J. Phycol. 4, 333–340.
Fraga, S., Gallager, S.M., Anderson, D.M., 1989. Chain-forming dinoflagellates: an
adaptation to red tides. In: Okaichi, T., Anderson, D., Nemoto, T. (Eds.), Red
Tides: Biology, Environmental Science and Toxicology. Elsevier, New York, pp.
281–284.
Gárate-Lizárraga, I., Bustillos-Guzmán, J., Morquecho, J., Lechuga-Déveze, C.,
2000. First outbreak of Cochlodinium polykrikoides in the Gulf of California.
Harmful Algae News, Intergovernmental Oceanographic Commission of
UNESCO, 21, 7 pp.
Gárate-Lizárraga, I., López-Corets, J.J., Bustillos-Guzmán, J.J., Hernández-Sandoval,
F., 2004. Blooms of Cochlodinium polykrikoides (Gymnodiniaceae) in the Gulf of
California, Mexico. Rev. Biol. Trop. 52.
Glibert, P., Anderson, D., Gentien, P., Granéli, E., Sellner, K., 2005. The global,
complex phenomena of harmful algal blooms. Oceanography 18, 136–147.
Gobler, C.J., Berry, D.L., Anderson, O.R., Burson, A., Koch, F., Rodgers, B.S., Moore, L.K.,
Goleski, J.A., Allam, B., Bowser, P., Tang, Y.Z., Nuzzi, R., 2008. Characterization,
dynamics, and ecological impacts of harmful Cochlodinium polykrikoides blooms
on eastern Long Island, NY, USA. Harmful Algae 7, 293–307.
84
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
Gobler, C.J., Burson, A.M., Tang, Y.Z., Mulholland, M.R. The role of nitrogenous
nutrients in the occurrence of the harmful dinoflagellate blooms caused by
Cochlodinium polykrikoides in New York estuaries. Harmful Algae, submitted for
publication.
Gobler, C.J., Renaghan, M.J., Buck, N.J., 2002. Impacts of nutrients and grazing
mortality on the abundance of Aureococcus anophagefferens during a New York
brown tide bloom. Limnol. Oceanogr. 47, 129–141.
Gomez-Villarreal, M.C., Martinez-Gaxiola, M.D., Peña-Manjarrez, J.L., 2008. Algal
blooms at Banderas, Bay, Mexico (2000–2001), from SeaWiFS-sensor-data. Rev.
Biol. Trop. 56, 1653–1664.
Gower, J., King, S., 2007. An Antarctic ice-related ‘‘superbloom’’ observed with the
MERIS satellite imager. Geophys. Res. Lett. 34, L15501 doi:15510.11029/
12007GL029638.
Gower, J., King, S., Borstad, G., Brown, L., 2005. Detection of intense plankton blooms
using the 709 nm band of the MERIS imaging spectrometer. Int. J. Remote Sens.
26, 2005–2012.
Guiry, M.D., Guiry, G.M., 2011. AlgaeBase. World-wide Electronic Publication.
National University of Ireland, Galway. , http://www.algaebase.org.
Guzmán, H.M., Cortes, J., Glynn, P.W., Richmond, R.H., 1990. Coral mortality
associated with dinoflagellate blooms in the eastern Pacific (Costa Rica and
Panama). Mar. Ecol. Prog. Ser. 60, 299–303.
Hallegraef, G.M., 1993. A review of harmful algal blooms and their apparent global
increase. Phycologia 32, 79–99.
Hallegraeff, G.M., 1992. Harmful algal blooms in the Australian region. Mar. Pollut.
Bull. 25, 186–190.
Hallegraeff, G.M., 2010. Ocean climate change, phytoplankton community
responses, and harmful algal blooms: a formidable predictive challenge. J. Phycol.
46, 220–235.
Han, H.-K., Kim, Y.-M., Lim, S.-J., Hong, S.-S., Jung, S.-G., Cho, H., Lee, W., Jin, E., 2010.
Enhanced efficacy of TD53, a novel algicidal agent, against the harmful algae via
the liposomal delivery system. Int. J. Pharm. 405, 137–141.
Hansen, P.J., 2011. The role of photosynthesis and food uptake for the growth of
marine mixotrophic dinoflagellates. J. Eukaryot. Microbiol. 58 (3), 203–214.
Hargraves, P.E., Maranda, L., 2002. Potentially toxic or harmful microalgae from the
northeast coast. Northeastern Nat. 9, 81–120.
Heisler, J., Glibert, P., Burkholder, J., Anderson, D., Cochlan, W., Dennison, W., Dortch,
Q., Gobler, C., Heil, C., Humphries, E., Lewitus, A., Magnien, R., Marshall, H.,
Sellner, K., Stockwell, D., Stoecker, D., Suddleson, M., 2008. Eutrophication and
harmful algal blooms: a scientific consensus. Harmful Algae 8, 3–13.
Ho, M.S., Zubkoff, P.L., 1979. The effects of a Cochlodinium heterolobactum bloom on
the survival and calcium uptake by larvae of the American oyster Crassostrea
virginica. In: Taylor, F., Seliger, H. (Eds.), Toxic Dinoflagellate Blooms. Elsevier,
North Holland, New York.
Holliday, D.V., Pieper, R.E., Kleppel, G.S., 1989. Determination of zooplankton size
and distribution with multifrequency acoustic technology. J. Cons. Int. Explor.
Mer. 46, 52–61.
Holmes, R., William, P., Eppley, R., 1967. Red water in La Jolla Bay, 1964–1966.
Limnol. Oceanogr. 12, 503–512.
Honda, A., Ishida, K., Miyamura, S., 1980. III. Yatsushiro Kaiiki, Yatsushiro Kai-1. In:
Fukuokaken, S.S., Sagaken, S.S., Nakgasaki, S.S., Kumamoto, S.S., Kagoshima,
S.S. (Eds.), Fisheries Agency, Suisan Shikenjhou. Kyushu Seiganiki Akashiwo
Yosatsu Chosa Houkokusho, (in Japanese), pp. 108–123.
Howard, M.D.A., Jones, A.C., Schnetzer, A., Countway, P.D., Tomas, C.R., Kudela, R.M.,
Hayashi, K., Chia, P., Caron, D.A. Quantitative real-time PCR for Cochlodinium
fulvescens (Dinophyceae), a potentially harmful dinoflagellate from California
coastal waters. J. Phycol., in press.
Huang, C., Dong, Q., 2000. Taxonomic and biological studies on causative organisms
from a large-scale red tide occurring in Pearl River Estuary in the spring, 1998.
Oceanol. Limnol. Sin./Haiyang Yu Huzhao 31, 233–238.
Humbert, J.F., Quiblier, C., Gugger, M., 2010. Molecular approaches for monitoring
potentially toxic marine and freshwater phytoplankton species. Anal. Bioanal.
Chem. 397, 1723–1732.
Imai, I., Kimura, S., 2008. Resistance of the fish-killing dinoflagellate Cochlodinium
polykrikoides against algicidal bacteria isolated from the coastal sea of Japan.
Harmful Algae 7, 360–367.
Imai, I., Yamaguchi, M., Hori, Y., 2006. Eutrophication and occurrences of harmful
algal blooms in the Seto Inland Sea, Japan. Plankton Benthos Res. 1, 71–84.
Iwataki, M., Hansen, G., Moestrup, O., Matsuoka, K., 2010. Ultrastructure of the
harmful unarmored dinoflagellate Cochlodinium polykrikoides (Dinophyceae)
with reference to the apical groove and flagellar apparatus. J. Eukaryot. Microbiol. 57, 308–321.
Iwataki, M., Kawami, H., Matsuoka, K., 2007. Cochlodinium fulvescens sp. nov.
(Gymnodiniales, Dinophyceae), a new chain-forming unarmored dinoflagellate
from Asian coasts. Phycol. Res. 55, 231–239.
Iwataki, M., Kawami, H., Mizushima, K., Mikulski, C.M., Doucette, G.J., Relox, J.R.,
Anton, A., Fukuyo, Y., Matsuoka, K., 2008. Phylogenetic relationships in the
harmful dinoflagellate Cochlodinium polykrikoides (Gymnodiniales, Dinophyceae) inferred from LSU rDNA sequences. Harmful Algae 7, 271–277.
Jeong, H.J., Kim, H.R., Kim, K.I., Kim, K.Y., Park, K.H., Kim, S.T., Yoo, Y.D., Song, J.Y.,
Kim, J.S., Seong, K.A., Yih, W.H., Pae, S.J., Lee, C.H., Huh, M.D., Lee, S.H., 2002.
NaOCl produced by electrolysis of natural seawater as a potential method to
control marine red-tide dinoflagellates. Phycologia 41, 643–656.
Jeong, H.J., Kim, J.S., Yoo, Y.D., Kim, S.T., Song, J.Y., Kim, T.H., Seong, K.A., Kang, N.S.,
Kim, M.S., Kim, J.H., Kim, S., Ryu, J., Lee, H.M., Yih, W.H., 2008. Control of the
harmful alga Cochlodinium polykrikoides by the naked ciliate Strombidinopsis
jeokjo in mesocosm enclosures. Harmful Algae 7, 368–377.
Jeong, H.J., Shim, J.H., Kim, J.S., Lee, Y.P.J., Lee Y., C.W., 1999. Feeding by the
mixotrophic thecate dinoflagellate Fragilidium cf. mexicanum on red-tide and
toxic dinoflagellates. Mar. Ecol. Prog. Ser. 176, 263–277.
Jeong, H.J., Yoo, Y.D., Kim, J.S., Kim, T.H., Kim, J.H., Kang, N.S., Yih, W., 2004.
Mixotrophy in the phototrophic harmful alga Cochlodinium polykrikoides (Dinophyceae): prey species, the effects of prey concentration, and grazing impact. J.
Eukaryot. Microbiol. 51, 563–569.
Jeong, H.J., Yoo, Y.D., Park, J.Y., Song, J.Y., Kim, S.T., Lee, S.H., Kim, K.Y., Yih, W.H.,
2005. Feeding by phototrophic red-tide dinoflagellates: five species newly
revealed and six species previously known to be mixotrophic. Aquat. Microb.
Ecol. 40, 133–150.
Jeong, J.-H., Jin, H.-J., Sohn, C., Suh, K.-H., Hong, Y.-K., 2000. Algicidal activity of
the seaweed Corallina pilulifera against red tide microalgae. J. Appl. Phycol. 12,
37–43.
Jeong, S.-Y., Ishida, K., Ito, Y., Okada, S., Murakami, M., 2003. Bacillamide, a novel
algicide from the marine bacterium, Bacillus sp. SY-1, against the harmful
dinoflagellate, Cochlodinium polykrikoides. Tetrahedron Lett. 44, 8005–8007.
Jiang, X., Tang, Y., Lonsdale, D.J., Gobler, C.J., 2009. Deleterious consequences of a red
tide dinoflagellate Cochlodinium polykrikoides for the calanoid copepod Acartia
tonsa. Mar. Ecol. Prog. Ser. 390, 105–116.
Jiang, X.D., Lonsdale, D.J., Gobler, C.J., 2010a. Density-dependent nutritional value of
the dinoflagellate Cochlodinium polykrikoides to the copepod Acartia tonsa.
Limnol. Oceanogr. 55, 1643–1652.
Jiang, X.D., Lonsdale, D.J., Gobler, C.J., 2010b. Grazers and vitamins shape chain
formation in a bloom-forming dinoflagellate, Cochlodinium polykrikoides. Oecologia 164, 455–464.
Jochens, A.E., Malone, T.C., Stumpf, R.P., Hickey, B.M., Carter, M., Morrison, R., Dyble,
J., Jones, B., Trainer, V.L., 2010. Integrated ocean observing system in support of
forecasting harmful algal blooms. Mar. Technol. Soc. J. 44, 99–121.
Ki, J.S., Han, M.S., 2008. Implications of complete nuclear large subunit ribosomal
RNA molecules from the harmful unarmored dinoflagellate Cochlodinium polykrikoides (Dinophyceae) and relatives. Biochem. Syst. Ecol. 36, 573–583.
Kim, C.-H., Cho, H.-J., Shin, J.-B., Moon, C.-H., Matsuoka, K., 2002. Regeneration from
hyaline cysts of Cochlodinium polykrikoides (Gymnodiniales, Dinophyceae), a
red tide organism along the Korean coast. Phycologia 41, 667–669.
Kim, C.-J., Kim, H.-G., Kim, C.-H., Oh, H.-M., 2007. Life cycle of the ichthyotoxic
dinoflagellate Cochlodinium polykrikoides in Korean coastal waters. Harmful
Algae 6, 104–111.
Kim, C.S., Lee, S.G., Lee, C.K., Kim, H.G., Jung, J., 1999. Reactive oxygen species as
causative agents in the ichthyotoxicity of the red tide dinoflagellate Cochlodinium polykrikoides. J. Plankton Res. 21, 2105–2115.
Kim, D., Kim, J.F., Yim, J.H., Kwon, S.K., Lee, C.H., Lee, C.K., 2008. Red to red – the
marine bacterium Hahella chejuensis and its product prodigiosin for mitigation
of harmful algal blooms. J. Microbiol. Biotechnol. 18, 1621–1629.
Kim, D., Oda, T., Muramatsu, T., Kim, D., Matsuyama, Y., Honjo, T., 2002. Possible
factors responsible for the toxicity of Cochlodinium polykrikoides, a red tide
phytoplankton. Comp. Biochem. Physiol. C 132, 415–423.
Kim, D.-I., Matsuyama, Y., Nagasoe, S., Yamaguchi, M., Yoon, Y.-H., Oshima, Y.,
Imada, N., Honjo, T., 2004. Effects of temperature, salinity, and irradiance on the
growth of the harmful red tide dinoflagellate Cochlodinium polykrikoides Margalef (Dinophyceae). J. Plankton Res. 26, 61–66.
Kim, E., Lee, H., Na, J.W.C., Kang, D., 2010. 5-MHz acoustic-backscatter measurements of Cochlodinium polykrikoides blooms in Korean coastal waters. ICES J.
Mar. Sci. 67, 1759–1765.
Kim, H., Kim, D., Lee, D., Park, C., Kim, H., 2001a. Limiting nutrients of Cochlodinium
polykrikoides red tide in Saryang Island coast by algal growth potential (AGP)
assay. J. Korean Fish. Soc. 34, 457–464.
Kim, H., Lee, C., Lee, S., Kim, H., Park, C., 2001b. Physico-chemical factors on the
growth of Cochlodinium polykrikoides and nutrient utilization. J. Korean Fish.
Soc. 34, 445–456.
Kim, H.G., 1998. Harmful algal blooms in Korean coastal waters focused on three
fish killing dinoflagellates. In: Lee, H., Lee, S., Lee, C. (Eds.), Harmful Algal
Blooms in Korea and China. NFRDI, Pusan, Korea.
Kim, H.G., 1997. Recent harmful algal blooms and mitigation strategies in Korea.
Ocean Res. (Seoul) 19, 185–192.
Kim, J.-S., Kim, J.-C., Lee, S., Lee, B.-H., Cho, K.Y., 2006. Biological activity of l-2azetidinecarboxylic acid, isolated from Polygonatum odoratum var. pluriflorum,
against several algae. Aquat. Bot. 85, 1–6.
Kim, M.J., Jeong, S.Y., Lee, S.J., 2008. Isolation, identification, and algicidal activity
of marine bacteria against Cochlodinium polykrikoides. J. Appl. Phycol. 20,
1069–1078.
Kim, S., Lee, H., Lee, Y., Yim, J., 2008. Mutant selection of Hahella chejuensis KCTC
2396 and statistical optimization of medium components for prodigiosin yieldup. J. Microbiol. 46, 183–188.
Kim, Y., Byun, Y., Kim, Y., Eo, Y., 2009. Detection of Cochlodinium polykrikoides
red tide based on two-stage filtering using MODIS data. Desalination 249,
1171–1179.
Kim, Y.S., Jeong, C.S., Seong, G.T., Han, I.S., Lee, Y.S., 2010. Diurnal vertical migration
Cochlodinium polykrikoides during the red tide in Korean coastal sea waters. J.
Environ. Biol. 31, 687–693.
Kofoid, C.A., Swezy, O., 1921. The Free Living Unarmoured Dinoflagellata. Memoirs
of the Univeristy of California, Berkeley.
Kraemer, G.P., Carmona, R., Chopin, T., Neefus, C., Tang, X., Yarish, C., 2004.
Evaluation of the bioremediatory potential of several species of the red alga
Porphyra using short-term measurements of nitrogen uptake as a rapid bioassay. J. Appl. Phycol. 16, 489–497.
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
Kudela, R., Ryan, J., Blakely, M., Lane, J., Peterson, T., 2008. Linking the physiology
and ecology of Cochlodinium to better understand harmful algal bloom events: a
comparative approach. Harmful Algae 7, 278–292.
Kudela, R.M., Seeyave, S., Cochlan, W.P., 2010. The role of nutrients in regulation and
promotion of harmful algal blooms in upwelling systems. Prog. Oceanogr. 85,
122–135.
Landsberg, J.H., 2002. The effects of harmful algal blooms on aquatic organisms. Rev.
Fish. Sci. 10, 113–390.
Larsen, J., Sournia, A., 1991. Diversity of heterotrophic dinoflagellates. In: Patterson,
D.J., Larsen, J. (Eds.), The Biology of Free-living Heterotrophic Dinoflagellates.
Clarendon Press, Oxford, pp. 313–332.
Lee, B.-K., Katano, T., Kitamura, S.-I., Oh, M.-J., Han, M.-S., 2008. Monitoring of
algicidal bacterium, Alteromonas sp. Strain A14 in its application to natural
Cochlodinium polykrikoides blooming seawater using fluorescence in situ hybridization. J. Microbiol. 46, 274–282.
Lee, C.K., Kim, H.C., Lee, S.-G., Jung, C.S., Kim, H.G., Lim, W.A., 2001. Abundance of
harmful algae, Cochlodinium polykrikoides, Gyrodinium impudicum and Gymnodinium catenatum in the coastal area of South Sea of Korea and their effects of
temperature, salinity, irradiance and nutrient on the growth in culture. J.
Korean Fish. Soc. 34, 536–544.
Lee, D.-K., 2008. Cochlodinium polykrikoides blooms and eco-physiological conditions in the South Sea of Korea. Harmful Algae 7, 318–323.
Lee, J.S., 1996. Bioactive components from red tide plankton Cochlodinium polykrikoides. J. Korean Fish. Soc. 29, 165–173.
Lee, M., Kim, B., Kwon, Y., Kim, J., 2009. Characteristics of the marine environment
and algal blooms in Gamak Bay. Fish. Sci. 75, 401–411.
Lee, M.O., Choi, J.H., Park, I.H., 2010. Outbreak conditions for Cochlodinium polykrikoides blooms in the southern coastal waters of Korea. Mar. Environ. Res. 70,
227–238.
Lee, Y.-J., Choi, J.-K., Kim, E.-K., Youn, S.-H., Yang, E.-J., 2008. Field experiments on
mitigation of harmful algal blooms using a Sophorolipid – yellow clay mixture
and effects on marine plankton. Harmful Algae 7, 154–162.
Lee, Y.S., Kim, J.D., Lim, W.A., Lee, S.G., 2009. Survival and growth of Cochlodinium
polykrikoides red tide after addition of yellow loess. J. Environ. Biol. 30, 929–932.
Lee, Y.S., Lee, S.Y., 2006. Factors affecting outbreaks of Cochlodinium polykrikoides
blooms in coastal areas of Korea. Mar. Pollut. Bull. 52, 626–634.
Mague, T.H., Friberg, E., Hughes, D.J., Morris, I., 1980. Extracellular release of carbon
by marine phytoplankton: a physiological approach. Limnol. Oceanogr. 25,
262–279.
Maldonado, D.J.C., 2008. Spectral properties and population dynamics of the
harmful dinoflagellate Cochlodinium polykrikoides (Margalef) in southwestern
Puerto Rico. Ph.D. Thesis. University of Puerto Rico, Mayagüez, Puerto Rico,
166 pp.
Margalef, R., 1961. Hidrografia y fitoplancton de un area marina de la costa
meridional de Puerto Rico. Inv. Pesq. 18, 76–78.
Marshall, H.G., 2009. Phytoplankton of the York River. J. Coast. Res. 57, 59–65.
Marshall, H.G., Egerton, T.A., Burchardt, L., Cerbin, S., Kokocinski, M., 2005. Long
term monitoring results of harmful algal populations in Chesapeake Bay and its
major tributaries in Virginia, U.S.A. Ocean. Hydrobiol. Stud. 34 (3), 35–41.
Matsuoka, K., Fukuyo, Y., 2000. Technical Guide for Modern Dinoflagellate Cyst
Study. IOC/WESTPAC-HAB, the University of Tokyo, Tokyo, 29 pp., 22 pls.
Matsuoka, K., Fukuyo, Y., 2002. Technical guide for modern dinoflagellate cyst
study. In: WESTPAC-HAB/WEATPAC/IOC, Japanese Society for the Promotion of
Science, p. 29.
Matsuoka, K., Iwataki, M., Kawami, H., 2008. Morphology and taxonomy of chainforming species of the genus Cochlodinium (Dinophyceae). Harmful Algae 7,
261–270.
Matsuoka, K., Mizuno, A., Iwataki, M., Takano, Y., Toshifumi, Y., Yoon, Y.H., Lee, J.B.,
2010. Seed populations of a harmful unarmored dinoflagellate Cochlodinium
polykrikoides Margalef in the East China Sea. Harmful Algae 9, 548–556.
Mikulski, C., Park, Y., Jones, K., Lee, C., Lim, W., Lee, Y., Scholin, C., Doucette, G., 2008.
Development and field application of rRNA-targeted probes for the detection of
Cochlodinium polykrikoides Margalef in Korean coastal waters using whole cell
and sandwich hybridiization formats. Harmful Algae 7, 347–359.
Montero, M.V., Bustamante, E.F., Guzman, J.C., Vargas, J.C., 2008. Harmful blooms by
noxious dinoflagellates in the Pacific coast of Costa Rica. Hydrobiologica 18, 15–23.
Morales-Blake, A., Cavazos-Guerra, C., Hernandez-Becerril, D., 2001. Unusual HABs
in Manzanillo Bay, Colima, Mexico. Harmful Algae News 22, 6.
Morse, R.E., Shen, J., Blanco-Garcia, J.L., Hunley, W.S., Fentress, S., Wiggins, M.,
Mulholland, M.R., 2011. Environmental and physical controls on the formation
and transport of blooms of the dinoflagellate Cochlodinium polykrikoides Margalef in the lower Chesapeake Bay and its tributaries. Estuar. Coast.
Mulholland, M.R., Morse, R.E., Boneillo, G.E., Bernhardt, P.W., Filippino, K.C., Procise,
L.A., Blanco-Garcia, J.L., Marshall, H.G., Egerton, T.A., Hunley, W.S., Moore, K.A.,
Berry, D.L., Gobler, C.J., 2009. Understanding causes and impacts of the dinoflagellate, Cochlodinium polykrikoides, blooms in the Chesapeake Bay. Estuar. Coast
32, 734–747.
Nagai, S., Nishitani, G., Sakamoto, S., Sugaya, T., Lee, C.K., Kim, C.H., Itakura, S.,
Yamaguchi, M., 2009. Genetic structuring and transfer of marine dinoflagellate
Cochlodinium polykrikoides in Japanese and Korean coastal waters revealed by
microsatellites. Mol. Ecol. 18, 2337–2352.
Nagayama, K., Shibata, T., Fujimoto, K., Honjo, T., Nakamura, T., 2003. Algicidal effect
of phlorotannins from the brown alga Ecklonia kurome on red tide microalgae.
Aquaculture 218, 601–611.
Neori, A., Chopin, T., Troell, M., Buschmann, A.H., Kraemer, G.P., Halling, C., Shpigel,
M., Yarish, C., 2004. Integrated aquaculture: rationale, evolution and state of the
85
art emphasizing seaweed biofiltration in modern mariculture. Aquaculture 231,
361–391.
National Fisheries Research Development Institute (NFRDI), 1998. Recent Red Tides
in Korea Coastal Waters, 2nd Ed. National Fisheries Research and Development
Institute, Pusan, Republic of Korea, 292 pp. (in Korean).
Officer, C.B., Smayda, T.J., Mann, R., 1982. Benthic filter feeding: a natural eutrophication control. Mar. Ecol. Prog. Ser. 9, 203–210.
Oh, M.Y., Lee, S.B., Jin, D.H., Hong, Y.K., Jin, H.J., 2010. Isolation of algicidal compounds from the red alga Corallina pilulifera against red tide microalgae. J. Appl.
Phycol. 22, 453–458.
Oh, S.J., Yoon, Y.H., Kim, D.-I., Shimasaki, Y., Oshima, Y., Honjo, T., 2006. Effect of light
quantity and quality on the growth of the harmful dinoflagellate Cochlodinium
polykrikoides Margalef (Dinophyceae). Algae 21, 311–316.
Okamura, K., 1916. Akashio ni Tsuite.([[nl]On red tides). Suisan Koushu Sikenjo
Kenkyu Hokoku 12, 26–41.
Onitsuka, G., Miyahara, K., Hirose, N., Watanabe, S., Semura, H., Hori, R., Nishikawa,
T., Miyaji, K., Yamaguchi, M., 2010. Large-scale transport of Cochlodinium
polykrikoides blooms by the Tsushima Warm Current in the southwest Sea of
Japan. Harmful Algae 9, 390–397.
Onoue, Y., Nozawa, K., 1989a. Separation of toxins from harmful red tides occurring
along the coast of Kagoshima Prefecture. In: Okaichi, T., Anderson, D.M., Nemoto, T. (Eds.), Red Tides, Biology, Environmental Science and Toxicology.
Elsevier, New York, pp. 371–374.
Onoue, Y., Nozawa, K., 1989b. Zinc-bound PSP toxins separated from Cochlodinium
red tide. In: Natori, S., Hashimoto, K., Ueno, Y. (Eds.), Mycotoxins and Phycotoxins’88. Elsevier, Amsterdam, pp. 359–366.
Onoue, Y., Nozawa, K., Kumanda, K., Takeda, K., Aramaki, T., 1985. Toxicity of
Cochlodinium’78 Yatsushiro occurring in Yatsushiro Sea. Nippon Suisan Gakkaishi 51, 147–151.
Orlova, T.Y., Morozova, T.V., Gribble, K.E., Kulis, D.M., Anderson, D.M., 2004. Dinoflagellate cysts in recent marine sediments from the east coast of Russia. Bot.
Mar. 47, 184–201.
Orlova, T.Y., Stonik, I.V., Shevchenko, O.G., 2009. Flora of planktonic microalgae of
Amursky Bay, Sea of Japan. Planktonology 35, 60–78.
Ou, L.-J., Zhang, Y.-Y., Li, Y., Wang, H.-J., Xie, X.-D., Rong, Z.-M., Lu, S.-H., Qi, Y.-Z.,
2010. The outbreak of Cochlodinium geminatum bloom in Zhuhai, Guangdong. J.
Trop. Oceanogr. 29, 57–61.
Palanisamy, S., Ahn, Y.-H., Ryu, J.-H., Moon, J.-E., 2005. Application of optical remote
sensing imagery for detection of red tide algal blooms in Korean waters. In: IEEE
International, IGARSS’05, vol. 3. pp. 1912–1915.
Park, G.-H., Lee, K., Koo, C.-M., Lee, H.-W., Lee, C.-K., Koo, J.-S., Lee, T., Ahn, S.-H., Kim,
H.-G., Park, B.-K., 2005. A sulfur hexafluoride-based Lagrangian study on
initiation and accumulation of the red tide Cochlodinium polykrikoides in
southern coastal waters of Korea. Limnol. Oceanogr. 50, 578–586.
Park, J.G., Jeong, M.K., Lee, J.A., Cho, K.-J., Kwon, O.-S., 2001. Diurnal vertical
migration of a harmful dinoflagellate, Cochlodinium polykrikoides (Dinophyceae) during a red tide in coastal waters of Namhae Island, Korea. Phycologia
40, 292–297.
Park, T.-G., Park, G.-H., Park, Y.-T., Kang, Y.-S., Bae, H.-M., Kim, C.-H., Jeong, H.-J., Lee,
Y., 2009. Identification of the dinoflagellate community during Cochlodinium
polykrikoides (Dinophyceae) blooms using amplified rDNA melting curve analysis and real-time PCR probes. Harmful Algae 8, 430–440.
Park, T.-G., Park, Y.-T., 2010. Detection of Cochlodinium polykrikoides and Gymnodinium impudicum (Dinophyceae) in sediment samples. Harmful Algae 9, 59–65.
Phlips, E.J., Badylak, S., Christman, M., Wolny, J., Brame, J., Garland, J., Hall, L., Hart, J.,
Landsberg, J., Lasi, M., Lockwood, J., Paperno, R., Scheidt, D., Staples, A., Steidinger, K., 2011. Scales of temporal and spatial variability in the distribution of
harmful algae species in the Indian River Lagoon, Florida, USA. Harmful Algae
10, 277–290.
Qi, D., Huang, Y., Wang, X., 1993. Toxic dinoflagellate red tide by a Cochlodinium sp.
along the coast of Fujian, China. In: Smayda, T., Shimizu, Y. (Eds.), Toxic
Phytoplankton Blooms in the Sea. Elsevier, Amsterdam.
Ramaiah, N., Paul, J.T., Fernandes, V., Raveendran, T., Raveendran, O., Sundar, D.,
Revichandran, C., Shenoy, D.M., Gauns, M., Kurian, S., Gerson, V.J., 2005. The
September 2004 stench off the southern Malabar coast – a consequence of
holococcolithophore bloom. Curr. Sci. 88, 551–554.
Rashed-Un-Nabi, M., Ee, L.S., Hoque, M.A., Sidik, M.J., Saleh, E., Anton A, 2010.
Effects of red tide on physico-chemical properties of water and phytoplankton assemblage in Sepanggar Bay, Sabah, Malaysia. Int. J. Ecol. Environ. Sci. 36,
245–251.
Richlen, M.L., Morton, S.L., Jamali, E.A., Rajan, A., Anderson, D.M., 2010. The catastrophic 2008–2009 red tide in the Arabian gulf region, with observations on
the identification and phylogeny of the fish-killing dinoflagellate Cochlodinium
polykrikoides. Harmful Algae 9, 163–172.
Roberts, R.J., 2001. The parasitology of teleosts. In: Roberts, R.J. (Ed.), Fish Pathology.
Elsevier, New York, pp. 254–296.
Robertson-Andersson, D., Potgieter, M., Hansen, J., Bolton, J., Troell, M., Anderson, R.,
Halling, C., Probyn, T., 2008. Integrated seaweed cultivation on an abalone farm
in South Africa. J. Appl. Phycol. 20, 579–595.
Rosales-Loessener, F., Matsuoka, K., Fukuyo, Y., Sanchez, E.H., 1996. Cysts of
harmful dinoflagellates found from Pacific coastal waters of Guatemala. In:
Yasumoto, T., Oshima, Y., Fukuyo, Y. (Eds.), Harmful and Toxic Algal Blooms.
International Oceanographic Commission of UNESCO, Paris.
Rubino, F., Belmonte, M., Caroppo, C., Giacobbe, M., 2010. Dinoflagellate cysts from
surface sediments of Syracuse Bay (Western Ionian Sea, Mediterranean). DeepSea Res. II 57, 243–247.
86
R.M. Kudela, C.J. Gobler / Harmful Algae 14 (2012) 71–86
Ryan, J., Fischer, A., Kudela, R., Gower, J., King, S., Marin Iii, R., Chavez, F., 2009.
Influences of upwelling and downwelling winds on red tide bloom dynamics in
Monterey Bay, California. Cont. Shelf Res. 29, 785–795.
Samson, J.C., Shumway, S.E., Weis, J.S., 2008. Effects of the toxic dinoflagellate,
Alexandrium fundyense on three species of larval fish: a food-chain approach. J.
Fish Biol. 72, 168–188.
Sannio, A., Luglie, A., Sechi, N., 1997. Potentially toxic dinoflagellates in Sardinia.
Plant Biosyst. 131, 73–78.
Saracino, O.D., Rubino, F., 2006. Phytoplankton composition and distribution along
the Albanian coast, South Adriatic Sea. Nova Hedwigia 83, 253–266.
Sasaki, H., Tanaka, A., Iwataki, M., Touke, Y., Siswanto, E., Tan, C.K., Ishizaka, J., 2008.
Optical properties of the red tide in Isahaya Bay, south-western Japan: influence
of chlorophyll a concentration. J. Oceanogr. 64, 511–523.
Saunders, G.W., Hill, D.A., Sexton, J.P., Andersen, R.A., 1997. Small-subunit ribosomal RNA sequences from selected dinoflagellates: testing classical evolutionary hypotheses with molecular systematic methods. Plant Syst. Evol. Suppl. 11,
237–260.
Schütt, F., 1895. Peridineen der Plankton-Expedition. Ergebn. Plankton–Expedition
der Humboldt-Stiftung 4 (M, a, A) 1–170, 127 pls.
Seaborn, D.W., Marshall, H.G., 2008. Dinoflagellate cysts within sediment collections from the southern Chesapeake Bay and tidal regions of the James, York,
and Rappahannock rivers. J. Sci. 59, 135–141.
Seong, K.A., Jeong, H.J., Kim, S., Kim, G.H., Kang, J.H., 2006. Bacterivory by cooccurring red-tide algae, heterotrophic nanoflagellates, and ciliates. Mar. Ecol.
Prog. Ser. 322, 85–97.
Shanmugam, P., Ahn, S.-H., Ram, P.S., 2008. SeaWiFS sensing of hazardous algal
blooms and their underlying mechanisms in shelf-slope waters of the Northwest Pacific during summer. Remote Sens. Environ. 112, 3248–3270.
Shao, P., Chen, Y.Q., Zhou, H., Yuan, J., Qu, L.H., Zhao, D., Lin, Y.S., 2004. Genetic
variability in Gymnodiniaceae ITS regions: implications for species identification and phylogenetic analysis. Mar. Biol. 144, 215–224.
Shin, K., Jang, M.-C., Jang, P.-K., Ju, S.-J., Lee, T.-K., Chang, M., 2003. Influence of food
quality on egg production and viability of the marine planktonic copepod
Acartia omorii. Prog. Oceanogr. 57, 265–277.
Silva, E.S., 1967. Cochlodinium heterolobactum n. sp.: structure and some cytophysiological aspects. J. Protozool. 14, 745–754.
Smayda, T.J., 1997. Harmful algal blooms: their ecophysiology and general relevance to phytoplankton blooms in the sea. Limnol. Oceanogr. 42, 1137–1153.
Smayda, T.J., 2007. Reflections on the ballast water dispersal – harmful algal bloom
paradigm. Harmful Algae 6, 601–622.
Smayda, T.J., 2008. Complexity in the eutrophication-harmful algal bloom relationship, with comment on the importance of grazing. Harmful Algae 8, 140–151.
Sogawa, K., Matsuda, M., Okutani, K., 1998a. Induction of apoptosis by a marine
microalgal polysaccharide in a human leukemic cell line. J. Mar. Biotechnol. 6,
241–243.
Sogawa, K., Sumida, T., Hamakawa, H., Yamada, T., Matsumoto, K., Matsuda, M., Oda,
H., Miyake, H., Tashiro, S., Okutani, K., 1998b. Inhibitory effect of a marine
microalgal polysaccharide on the telomerase activity in K562 cells. Res. Commun. Mol. Pathol. Pharmacol. 99, 259–265.
Sohn, M.H., Seo, K.W., Choi, Y.S., Lee, S.J., Kang, Y.S., Kang, Y.S., 2011. Determination
of the swimming trajectory and speed of chain-forming dinoflagellate Cochlodinium polykrikoides with digital holographic particle tracking velocimetry.
Mar. Biol. 158, 561–570.
Song, Y.-C., Sivakumar, S., Woo, J.-H., Ko, S.-J., Hwang, E.-J., Jo, Q., 2010. Removal of
Cochlodinium polykrikoides by dredged sediment: a field study. Harmful Algae 9,
227–232.
Stoecker, D., Tillmann, U., Granéli, E., 2006. Phagotrophy in harmful algae. In:
Granéli, E., Turner, J.T. (Eds.), Ecology of Harmful Algae, vol. 189. SpringerVerlag, Berlin, pp. 177–187.
Stumpf, R.P., Tomlinson, M.C., 2005. Remote sensing of harmful algal blooms.
Remote Sens. Coast. Aquat. Environ. 7, 277–296.
Suh, Y.S., Kim, J.H., Kim, H.G., 2000. Relationship between sea surface temperature
derived from NOAA satellites and Cochlodinium polykrikoides red tide occurrence in Korea coastal waters. J. Korean Environ. Sci. 9, 215–221.
Sullivan, J.M., Swift, E., Donaghay, P., Rines, J.E.B., 2003. Small-scale turbulence
affects the division rate and morphology of two red-tide dinoflagellates. Harmful Algae 2, 183–199.
Sun, X.-X., Choi, J.-K., Kim, E.-K., 2004a. A preliminary study on the mechanism of
harmful algal bloom mitigation by use of sophorolipid treatment. J. Exp. Mar.
Biol. Ecol. 304, 35–49.
Sun, X.-X., Lee, Y.-J., Choi, J.-K., Kim, E.-K., 2004b. Synergistic effect of sophorolipid
and loess combination in harmful algal blooms mitigation. Mar. Pollut. Bull. 48,
863–872.
Sunda, W.G., Graneli, E., Gobler, C.J., 2006. Positive feedback and the development
and persistence of ecosystem disruptive algal blooms. J. Phycol. 42, 963–974.
Tang, Y.Z., Gobler, C., 2009a. Cochlodinium polykrikoides blooms and clonal isolates
from the northwest Atlantic coast cause rapid mortality in larvae of multiple
bivalve species. Mar. Biol. 156, 2601–2611.
Tang, Y.Z., Gobler, C.J., 2009b. Characterization of the toxicity of Cochlodinium
polykrikoides isolates from Northeast US estuaries to finfish and shellfish.
Harmful Algae 8, 454–462.
Tang, Y.Z., Gobler, C.J., 2010. Allelopathic effects of Cochlodinium polykrikoides
isolates and blooms from the estuaries of Long Island, New York, on cooccurring phytoplankton. Mar. Ecol. Prog. Ser. 406, 19–31.
Tang, Y.Z., Gobler, C.J., 2011. The green macroalga, Ulva lactuca, inhibits the growth
of seven common harmful algal bloom species via allelopathy. Harmful Algae
10, 480–488.
Terenko, L., 2005. New dinoflagellate (Dinoflagellata) species from the Odessa Bay of
the Black Sea. Ocean. Hydrobiol. Stud. 34, 205–216.
Tomas, C.R., Smayda, T.J., 2008. Red tide blooms of Cochlodinium polykrikoides in a
coastal cove. Harmful Algae 7, 308–317.
Tort, M.J., Jennings-Bashore, C., Wilson, D., Wooster, G.A., Bowser, P.R., 2002.
Assessing the effects of increasing hydrogen peroxide dosage on rainbow
trout gills utilizing a digitized scoring methodology. J. Aquat. Anim. Health 14,
95–103.
Trainer, V.L., Pitcher, G.C., Reguera, B., Smayda, T.J., 2010. The distribution and
impacts of harmful algal bloom species in eastern boundary upwelling systems.
Prog. Oceanogr. 85, 33–52.
Vargas-Montero, M., Freer, E., Jimenez-Montealegre, R., Guzman, J., 2004. Extensive
blooms due to Cochlodinium polykrikoides: new to Costa Rica. Harmful Algae
News 26, 7.
Vargas-Montero, M., Freer, E., Jimenez-Montealegre, R., Guzman, J.C., 2006. Occurrence and predominance of the fish killer Cochlodinium polykrikoides on the
Pacific coast of Costa Rica. Afr. J. Mar. Sci. 28, 215–217.
Verity, P.G., 2010. Expansion of potentially harmful algal taxa in a Georgia Estuary
(USA). Harmful Algae 9, 144–152.
Vershinin, A.O., Moruchkov, A.A., Leighfield, T., Sukanova, I., Kamnev, A.N., Pankov,
S., Morton, S.L., Ramsdell, J.S., 2005. Potential toxic algae in northeast Black Sea
coastal phytoplankton in 2000–2002. Okeanologiya 45, 240–248.
Vershinin, A.O., Moruchkov, A.A., Sukanova, I., Kamnev, A.N., Pankov, S., Morton, S.L.,
Ramsdell, J.S., 2004. Seasonal changes in phytoplankton in the area of Cape
Bolshoi Utrish off northern Caucasian coast in the Black Sea 2001–2002. Russ.
Acad. Sci. Oceanol. 44, 379–387.
Viques, R., Hargraves, P.E., 1995. Annual cycle of potentially harmful dinoflagellates
in the Gulfo de Nicoya. Costa Rica Bull. Mar. Sci. 57, 467–475.
Whyte, J.N.C., Haigh, N., Ginther, N.G., Keddy, L.J., 2001. First record of blooms of
Cochlodinium sp. (Gymnodiniales, Dinophyceae) causing mortality to aquacultured salmon on the west coast of Canada. Phycologia 40, 298–304.
Yamasaki, Y., Nagasoe, S., Matsubara, T., Shikata, T., Shimasaki, Y., Oshima, Y., Honjo,
T., 2007. Growth inhibition and formation of morphologically abnormal cells of
Akashiwo sanguinea (Hirasaka) G. Hansen et Moestrup by cell contact with
Cochlodinium polykrikoides Margalef. Mar. Biol. 152, 157–163.
Yamatogi, T., Maruta, H., 2002. Occurrence of Cochlodinium polykrikoides red tide
and its growth characteristics in Imari Bay in 1999. Bull. Nagasaki Prefect. Inst.
Fish. 28, 21–26.
Yamatogi, T., Sakaguchi, M., Iwataki, M., Matsuoka, K., 2005. Seasonal occurrence
and growth characteristics of a harmful dinoflagellate Cochlodinium polykrikoides in Usuka Bay. Jpn. J. Phycol. 53, 229–235.
Yamatogi, T., Sakaguchi, M., Iwataki, M., Matsuoka, K., 2006. Effects of temperature
and salinity on the growth of four harmful red tide flagellates occurring in
Isahaya Bay in Ariake Sound, Japan. Nippon Suisan Gakkaishi 72, 160–168.
Yang, Y.-F., Fei, X.-G., Song, J.-M., Hu, H.-Y., Wang, G.-C., Chung, I.K., 2006. Growth
of Gracilaria lemaneiformis under different cultivation conditions and its
effects on nutrient removal in Chinese coastal waters. Aquaculture 254,
248–255.
Yoo, D.Y., Jeong, H.J., Kim, M.S., Kang, N.S., Song, J.Y., Shin, W., Kim, K.Y., Lee, K., 2009.
Feeding by phototrophic red-tide dinoflagellates on the ubiquitous marine
diatom Skeletonema costatum. J. Eukaryot. Microbiol. 56, 413–420.
Yuki, K., Yoshimatsu, S., 1989. Two fish-killing species of Cochlodinium from
Harima-Nada, Seto Inland Sea Japan. In: Okaichi, T., Anderson, D., Nemoto, T.
(Eds.), Red Tides: Biology, Environmental Science, and Toxicology. Elsevier,
New York, pp. 451–454.
Zhou, Y., Yang, H., Hu, H., Liu, Y., Mao, Y., Zhou, H., Xu, X., Zhang, F., 2006.
Bioremediation potential of the macroalga Gracilaria lemaneiformis (Rhodophyta) integrated into fed fish culture in coastal waters of north China. Aquaculture 252, 264–276.