The “harmful algae and algal toxins in coastal waters of China: investigation and database” project*

Tian YAN 1, 2, 3, **

1 CAS Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences,Qingdao 266071, China

2 Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266237, China

3 Center for Ocean Mega-Science, Chinese Academy of Sciences, Qingdao 266071, China

Abstract The “harmful algae and algal toxins in coastal waters of China: investigation and database”project (HAATC) is funded (US$2.3 million) by the Ministry of Science and Technology of China for 60 months starting on the 1 st Jan 2019. Since the 1990s, national projects and other related work have greatly improved harmful algal bloom (HAB) research in China. However, the existing research on HABs is often limited to specif ic sea areas or specif ic algal bloom disasters, which is insuffi cient for understanding the longterm evolution of HAB occurrence under the inf luence of intensif ied human activity and global change. The HAATC project intends to determine the overall distribution of marine harmful microalgae and macroalgae,and marine algal toxins (in phytoplankton and shellf ish) through a large-scale investigation in the coastal waters of China and continuous investigation in key sea areas. It also aims to observe biotic and abiotic processes during harmful red tides and green tides, and establish an open database and query platform of HABs in China. The research consortium, led by chief scientist Prof. Tian YAN, includes approximately 100 scientists and students. More than 2 years of work have shown the increased distribution of HAB species (including cysts) and algal toxins in China’s coastal waters, the increased occurrence of macroalgae blooms (mainly by Ulva prolifera and Sargassum horneri) in the Southern Yellow Sea, increased amount of causative species being identif ied due to the use of molecular technologies in taxonomy, and new records of algal toxins such as azaspiracid, cycloimines and neurotoxins β-N-methylamino-l-alanine have been detected, which complicates the research in algal toxins and toxicology. Eventually, the project will provide systematic basic data and techniques for the rapid and accurate identif ication of harmful algae species to support the research, monitoring, and control of harmful algal blooms, as well as seafood safety management in China.

Keyword: harmful algae; algal toxins; coastal waters of China; database

1 INTRODUCTION

Toxic and harmful algal blooms have frequently occurred and pose a serious problem in the marine environment. They were previously reported to have intensif ied in coastal regions globally in recent decades(Anderson et al., 2012). While recent statistical analysis has thrown doubt of previous assumptions of harmful algal bloom (HAB) increases at a global scale (Hallegraeff et al., 2021), rapid economic development and associated environmental impacts in Chinese waters, including benthos degradation (Xu et al., 2017) and increase in giant jellyf ish blooms(Sun et al., 2015) suggests that such doubt on HAB increase should not yet be expressed for Chinese waters. Off shore HABs mainly include red tides(referred to as microalgal blooms here on in), brown tides caused by picoalgaeAureococcussp., green tides caused by green macroalgae and golden tides caused by brown macroalgae (Cosper et al., 1990; Fletcher,1996; Anderson, 1997b; Smetacek and Zingone,2013). HABs are an algal bloom phenomenon that cause harm through massive proliferation or toxin contamination (Kudela et al., 2017). They have been increasing along the coast, and have had serious social and economic impacts in China over recent decades(Li et al., 2021). Moreover, algal toxins produced by toxic algae threaten food safety through the food chain, and increasing poisoning incidents such as paralytic shellf ish poisoning (PSP) and diarrhetic shellf ish poisoning (DSP) have been reported recently(Yan et al., 2002; Hallegraeff , 2021). Shellf ish exports to the EU were banned for 19 years until 2018 due to algal toxin contamination problems, resulting in serious economic losses. HAB disaster occurrence has extended along the coast of China from the south to the north, endangering human health, coastal tourism,off shore aquaculture, and nuclear power safety(Wang et al., 2021b; Gu et al., 2022). Furthermore,HABs have become a signif icant challenge for recent development strategies, such as the construction of“Healthy China” and “Beautiful China”, as well as the protection of “Blue Granary” and “Ecological Security”.

Chinese scientists started to study the problem of HABs during the 1970s. From the 1990s, China successively established several national HAB projects: two major National Natural Science Foundation of China projects in the 1990s led by Prof. Yuzao Qi, two 973 Plan projects led by Prof.Mingjiang Zhou since 2000, and two special Research and Development projects for red and green tides starting from 2017. Each of these national projects focused on the causes and control mechanisms of HABs in specif ic sea areas along the Chinese coast,such as Daya Bay, Jiaozhou Bay, Changjiang estuary,and the Beibu Gulf. For national red tide monitoring,19 zones, mainly in the estuaries and bays along the coast, were established by the State Oceanic Administration (SOA, now Ministry of Natural Resources) in China.

Through previous project implementation and other investigations, the causes, impacts, and mechanism of mitigation of major algal bloom disasters (such as blooms caused byProrocentrumdonghaiense,Kareniamikimotoi,Phaeocystisglobasa, andUlvaprolifera), as well as the regional distribution pattern of typical harmful algae and algal toxins in coastal waters, have been reviewed (Yan et al., 2022). However, general background data for the whole off shore region of China remains insuffi cient,and basic investigative project support is urgently required.Here, we summarize the main challenges related to harmful algae and algal toxins in China, and the“Harmful Algae and Algal Toxins in Coastal Waters of China: Investigation and Database” (HAATC)project, including its latest progress.

2 INCREASING MARINE DISASTERS RELATED TO HABS AND ALGAL TOXINS IN CHINA

Over the past 20 years, algal blooms caused by toxic and harmful algae in China’s coastal waters have been increasing (He et al., 2021). Toxic microalgae,such asAlexandriumsp.,Dinophysissp. andPseudonitzschiasp., threatening human life and health have attracted much attention in China, since they produce various algal toxins aff ecting seafood safety; Other harmful algae, such asK.mikimotoi,Aureococcusanophageff erens, andU.prolifera, although without any known human toxins, have impacted marine aquaculture, ecology, and environments in China through large-scale, high biomass blooms and the generation of other toxic substances (Xiao et al.,2021; Gu et al., 2022). To date, the algal blooms caused by toxic and harmful algae in China’s off shore waters have evolved, as described below:

Marine microalgae blooms in China are increasing in terms of both number of species, and toxic forms.Brown tides, which have previously only occurred in the United States and South Africa, have started to occur in the Bohai Sea of China almost annually, and have caused economic losses involving hundreds of millions of yuan in the scallop mariculture between 2007 and 2014 (Zhang et al., 2012).K.mikimotoiblooms, which had already caused a large number of f ish deaths worldwide, had a devastating impact on the abalone mariculture industry in Fujian in 2012,causing about US$ 300 million economic losses(MNR, 2012). The f ish-killingP.globosahas posed a major threat to the safety of the cold source system of the Guangxi nuclear power plant by blocking the f iltration facilities in recent years (Wang et al., 2021b).The emergence of newly-recorded algae species and new harmful forms in coastal waters increases the necessity of obtaining knowledge about background algal species, and techniques for identif ications and detections to support the early monitoring and warning of HABs.

Macroalgal bloom disasters in China’s coastal waters have shown an increasing trend. A massiveU.proliferagreen tide in the South Yellow Sea has occurred annually since 2007, resulting in more than US$ 15 million economic losses each year(Zhou et al., 2015). Moreover, aSargassumhornerigolden tide appeared off shore at the end of 2016,and impacted the area of laver culture in the shoal of northern Jiangsu for the f irst time (Xing et al.,2017). At that time, the laver industry in Yancheng and Nantong alone suff ered a direct loss of as much as US$ 75 million (MNR, 2017). Furthermore, a cooccurrence of green, golden, and red tides has been observed during the spring and summer 2017 in the Southern Yellow Sea, which is relatively rare along coasts globally (Kong et al., 2018). Compared with microalgal blooms, the social and economic losses associated with macroalgal blooms are greater due to the higher biomass and the large f loating algae,which can be transported with ocean currents causing disasters in diff erent places, making their prediction and prevention diffi cult (Xiao et al., 2020).

Marine algal toxins in China threaten food safety and human health. Recently, there has been a signif icant increase in seafood poisoning incidents caused by algal toxins (Yan et al., 2022). For example,PSP incidents were reported in Fujian Province in June 2017; more than 30 poisoned patients developed symptoms, such as dizziness, vomiting and numbness of limbs, after ingestion of contaminated mussels in Shishi (Quanzhou) and Longhai and Zhangpu(Zhangzhou) (Fan et al., 2021). Another PSP incident was caused by the consumption of purple mussels in Qinhuangdao City at the end of April 2016, where nine people were hospitalized (Yu and Luo, 2016).At the end of May 2011, more than 200 people were hospitalized for DSP caused by purple mussels in Ningde (Fujian) and Ningbo (Zhejiang) (Li et al.,2012). Since the poisoning symptoms of algal toxins are easily confused with other diseases, a considerable number of algal toxin poisoning events have not been identif ied or counted. Toxin events can cause market panic, a large amount of seafood has been destroyed or banned for sale, and earnings of foreign exchange has been severely and repeatedly impacted (Yan et al., 2022). In recent years, some new records of algal toxins, such as azaspiracid, cycloimines and neurotoxins β-N-methylamino-l-alanine have often been detected in shellf ish in China (Gu et al., 2013;Liu et al., 2019; Wang et al., 2021a).

In summary, the issues of HAB and algal toxins in China’s off shore waters are becoming increasingly complex and diverse. There is therefore an urgent need to further investigate harmful algae and algal toxins in coastal China.

3 INTERNATIONAL HAB RESEARCH

HABs have long been an important marine ecological environment problem and frontier issue in international marine scientif ic research. The Intergovernmental Oceanographic Commission(IOC) of the United Nations Educational, Scientif ic and Cultural Organization (UNESCO) and the Scientif ic Committee on Oceanic Research (SCOR)have jointly supported two international plans for HABs, the “Global Study of Harmful Algal Bloom Ecology and Oceanography” from 1998 to 2013 and the “Global Change and Harmful Algal Bloom Plan”launched in 2016 (GEOHAB, 2001; Kudela et al.,2017). The new plan advocates focusing attention on the occurrence of HABs, and the evolution of algal disasters under the accelerated impacts of global climate change and human activities. Harmful algae and algal toxins have always been one of the core issues of international marine research.

3.1 Progress in research and technology

In the study of harmful algae in off shore waters,the identif ication and classif ication of algae species forms the basis of all research and management.Researchers have performed extensive research and sorting relating to their taxonomy and geographic distribution. Since the 1990s, based on the morphological identif ication of traditional light and electron microscopy, molecular markers have been gradually used for algae species identif ication(Scholin and Anderson, 1994). Some algal species have been screened for genetic information in order to distinguish between subspecies and geographical populations, which has been convenient and has led to new understanding for the classif ication of algae species (Costas, 1997). Therefore, the classif ication of some microalgae and macroalgae has changed.For example, theKareniafamily was established in 2006 based on morphological characteristics,ribosomal large subunit DNA sequence, and pigment composition. Its species previously were assigned to GymnodinialesGymnodinium, and a few species belonged toGyrodiniumandKarenia(Bergholtz et al., 2006; Wang et al., 2017). The phylogenetic tree of theAlexandriumcomplex species was established based also on the sequence of large subunits. The species complex was divided into f ive distinct groups.Group I and group Ⅳ are toxic strains, while groupsⅡ, Ⅲ, and V are nontoxic strains (Wang et al., 2014).Enteromorphawas classif ied intoUlvabased on nuclear-encoded and chloroplast-encoded ribulosebisphosphate carboxylase gene sequences (Hayden et al., 2003).

In recent years, DNA barcoding technology has become an important technical support in taxonomy and biodiversity research (Saunders, 2005). In contrast to animals, it is more diffi cult to determine a single bar code gene in plants (Fazekas et al., 2009). Similarly, a unif ied bar code gene suitable for algae has not been found at present. However, for diatoms, red algae,brown algae, and green algae, DNA bar code gene combinations suitable for this category have been determined through extensive comparison (Saunders and McDevit, 2012). In addition, in the application of the amplicon high-throughput sequencing method for the investigation of genetic diversity of marine phytoplankton, the species of microalgae identif ied include common toxic and harmful species, as well as many species that are diffi cult to identify through traditional investigation methods, highlighting the technical advantages (Xiao et al., 2014). There remains much diversity in harmful microalgae to be discovered.

Scientists from various countries have performed a large amount of research into the distributions of harmful algae and algal toxins in diff erent sea areas(Meyer et al., 1928; Sournia, 1995; Yan et al., 2022).The United States and some European countries have determined relatively clear distribution patterns of the main toxic and harmful microalgae and algal toxins in diff erent sea areas, such asAlexandriumand its paralytic shellf ish toxins (PSTs) in the Northeast sea area of the United States (Anderson, 1997a),Kareniabrevisand its neurotoxic shellf ish toxins (NSTs)in the southeast sea area of the United States (Soto et al., 2018), andPseudo-nitzschiasp. and its amnesic shellf ish toxins (ASTs) along the West coast of the US(McCabe et al., 2016), providing a basic background for in-depth research and management. International research on dormant algal cells, especially dinof lagellate cysts, began in the 1960s (Wall and Dale,1966). The United States, Europe, Japan, and other countries have carried out surveys of dinof lagellate cysts in sediments (Wall and Dale, 1968; Matsuoka,1999; Dale, 2001). Recently, scientists from Canada,the United Kingdom, Belgium, Germany, and France have jointly investigated dinof lagellate cysts in the sediments of the Black Sea corridor (Mudie et al.,2017).

There are approximately 300 algal species that can cause HABs in the coastal sea, of which about 80 kinds are toxic (Hallegraeff , 2003). According to the intermediate vectors and toxic symptoms of algal toxins, the main algal toxins can be divided into NSTs, PSTs, ASTs, diarrhetic shellf ish toxins (DSTs)and ciguatera f ish toxins (CFTs) (Hallegraeff , 2003).Among them, PSTs and DSTs are the two most widely distributed and harmful algal toxins. They have been listed as routine detection items in the United Kingdom, the United States, Japan, South Korea, and China. According to statistics, more than 200 types of algal toxins and their derivatives have been found(Gerssen et al., 2011). Algal toxins can accumulate in f ilter feeding organisms, resulting in potential risks to the food safety of aquatic products (Hallegraeff ,1993). Worldwide, human poisonings and even death,are occasionally caused by algal toxins. There are about 60 000 poisoning incidents in the world every year related to marine algal toxins, mainly PSP and DSP (Gill et al., 2003). At present, the international detection and analysis methods of algal toxins mainly include biological test methods, chemical analysis methods and other methods (Stewart and McLeod,2014). Among these, the mouse method is the most commonly used biological method. Although its operation is simple, its sensitivity and accuracy are poor. Chemical analysis methods can provide accurate qualitative and quantitative analysis of algal toxins, but need to be calibrated against standard toxins. The main chemical analysis methods include high performance liquid chromatography, thin layer chromatography,capillary electrophoresis, high performance liquid chromatography-mass spectrometry and nuclear magnetic resonance. In addition, the detection methods of algal toxins also include protein phosphatase inhibition test, cytotoxicity test, immunological detection technology, and the biosensor method,amongst others (Carmichael and An, 1999; Heresztyn and Nicholson, 2001; Pierce and Kirkpatrick, 2001).

3.2 HAB guidelines and databases

The international community attaches great importance to the basic research methods on harmful algae. The manual on harmful marine microalgae(IOC manuals and guide No. 33) and its revision were published in 1995 and 2004, respectively. The manual, illustrating sampling, identif ication, toxin analysis, and monitoring management, was prepared by 46 well-known scientists with the support of the IOC of UNESCO.

To improve basic research and better protect the ecological environment, long-term preservation of toxic and harmful algae species resources has been carried out. The National Center for Marine Algae and Microbiota (formerly National Center for Culture of Marine Phytoplankton) of the United States has preserved more than 2 000 species of microalgae.Countries such as Australia and Canada have built algae banks involving thousands of strains. Japan has isolated more than 1 000 algal species through the project of “Earth Research Renewal Technology Plan”, and established the algal species bank of the National Institute of Environment of Japan.

During this century, international marine science organizations have begun to pay attention to the collection of harmful algal blooms and algal toxin pollution data. The 15thWorking Group of the North Pacif ic Marine Science Organization (PICES)summarized HABs in Japan, China, South Korea,Russia, Canada, the United States, and Mexico, and published a scientif ic report (Taylor and Trainer,2002) on HABs in the North Pacif ic in 2002.

Recently, databases and internet information platforms have been used to collect and release HAB information. In the action plan for the environmental protection, management and development of the Northwest Pacif ic Marine and coastal area of the United Nations Environment Program (Northwest Pacif ic Action Plan), the third working group of the Special Monitoring and Coastal Environmental Assessment Regional Activity Centre developed an integrated website related to HABs of Northwest Pacif ic Action Plan members in China, Japan, South Korea, and Russia (http://www.cearac-project.org/HAB_Integrated_Website/). It publishes information and data on the distribution of major harmful algae,algal bloom events, and toxin distribution in the aforementioned countries.

In 2009, nine internationally renowned microalgae classif ication experts established a toxic and harmful microalgae classif ication information platform (http://www.marinespecies.org/hab/index.php). At present,176 toxic and harmful species have been collected and updated, and species morphological information,photographs, classif ication, and simple toxin information have been listed. This website is mainly used for the species identif ication of toxic and harmful microalgae, clarif ication of classif ication information,change of name, and accurate classif ication.

The “Toxic and Harmful Microalgae of the World Ocean” published in 2016, updated the list of toxic and harmful phytoplankton species in the international database (Lassus et al., 2016). This included location,cell density, toxin level in HAB outbreak areas,extraction and analysis protocols of harmful algal active substances and toxins, and the geographical distribution of typical harmful algae and their possible distribution trend in the past 30 years. HAB outbreaks in European countries have shown a downward trend due to intensive monitoring and management, as well as global change and environmental restoration programs (https://ec.europa.eu/info/research-andinnovation/research-area/environment/oceans-andseas/eu-marine-strategy-framework-directive_en).

4 NATIONAL PROGRAMS OF HARMFUL ALGAE AND ALGAL TOXIN RESEARCH IN CHINA

Since the 1990s, the National Natural Science Foundation of China has established two major funds successively, “Research on the occurrence mechanism of red tides in the southeast coast of China” and“Research on the dynamics and control mechanism of harmful red tides in typical coastal aquaculture areas of China”. Since 2002, the Ministry of Science and Technology has successively organized and implemented two National Key Basic Research Program of China (973 Program); “Ecology and Oceanology, prediction and control of harmful red tides in China’s off shore” and “Evolution mechanism of algal bloom disaster and ecological safety in China’s off shore”, and two recently launched research and development projects of harmful algal blooms;“U.proliferagreen tide formation mechanism and comprehensive prevention and control technology”and “formation mechanism, monitoring, prediction and evaluation prevention and control technology of disaster causing red tide in China’s off shore”. In 2004,the SOA established 19 red tide monitoring areas nationwide. These projects and other related work have eff ectively improved HAB research in China.

Chinese researchers have obtained experience in the classif ication of phytoplankton and macroalgae;therefore, the list of diatoms and dinof lagellates has been continuously updated in the books of“Chinese Algae Records”, “Marine Species and Their Distributions in China’s Seas”, and “Marine Organisms in China” (Huang, 2008). Light and electron microscope photographs of algae are included in the books of “Dinof lagellates in the South China Sea”(Lin and Zhou, 1993) and “Dinof lagellates in China’s Sea” (Yang et al., 2014). The species and geographic distribution information of some disaster causative macroalgae in China’s coastal areas are included in “China’s Economic Algae Chronicle” (IOCAS,1962), “China’s Yellow and Bohai Sea Algae” (Zeng,2009), and “Volume IV of China’s Algae Chronicle”(Ding, 2013). However, these catalogues and maps were not complied from the perspective of toxic and harmful algae.

The “Atlas of red tide organisms in China’s coastal waters” published in 2004, and “The investigation and evaluation of red tide disasters in China (1933–2009)” reported the status of red tide organisms, red tide events, environmental factors, and algal toxins in some sea areas in China, and included pictures of red tide organisms. The SOA reports on the red tides and green tides in the bulletin of China’s Marine Environmental Quality and the Bulletin of China’s Marine Disasters annually. This includes the occurrence times and cumulative area of red tides in each sea area every year, the total monthly area of red tides, the dominant species and timings of red tide outbreaks, a brief description of the dominant species and maximum area, start and end times of some largescale red tide outbreaks, and the distribution and coverage area of green tides (MNR, 2009–2020).Chinese researchers have isolated toxic and harmful algae species from China’s coastal waters,and maintained a collection of algal cultures including microalgae and macroalgae in their respective laboratories for HAB research.

5 OVERALL DATA ON HARMFUL ALGAE AND ALGAL TOXINS IN CHINA’S OFFSHORE WATERS ARE URGENTLY NEEDED

There has been a considerable amount of research into the biology, ecology, and oceanography of HABs to date; however, it is often limited to specif ic sea areas or specif ic algal bloom disasters (Qi et al., 2017;Zhang et al., 2019; Li et al., 2021). Data on toxic and harmful algae, and algal toxins in China’s off shore are insuffi cient for understanding the distribution pattern, seafood safety risk, and pattern shift under the intensif ied inf luence of human activity and global climate change.

The HAATC project aims to obtain basic data on HAB species (microalgae and macroalgae, and cysts) and algal toxin distribution in China’s off shore waters, and the temporal and spatial dynamic changes of HAB species, algal toxins, and environmental factors in typical HAB zones. It intends to build a multi-information open platform of basic scientif ic data to support further HAB research, monitoring,and seafood safety management in China’s off shore waters. It will also be useful for understanding the long-term evolution of HAB occurrence under the inf luence of intensif ied human activity and global change.

6 HAATC IMPLEMENTATION PLAN

The HAATC research plan objectives are to:(1) investigate the large-scale distribution of toxic and harmful microalgae, and cysts in China’s off shore waters, and their temporal and spatial dynamic changes in key aquaculture areas and red tide areas;(2) investigate the overall distribution of algal toxins in shellf ish in China’s coastal waters, and their temporal and spatial distribution in key aquaculture areas and red tide areas; (3) investigate the distribution of disaster causative macroalgae along the coastline and in green tide areas of China; (4) collect environmental data from typical HAB zones; (5) collect typical harmful algae isolates, and study the morphological,genetic, pigment, and toxicity data of each species; and(6) establish an open database and multi information query platform of HABs.

The overall distribution of more than 100 HAB species and their cysts (if any), and 20 algal toxins will be studied in more than 50% of the waters offthe coast of China, as well as that of more than 20 macroalgae species along the coast. The temporal and spatial variation of harmful algae and algal toxins(both in phytoplankton and shellf ish), and biotic and abiotic factors will be collected under bloom dynamics in eight typical aquaculture areas and HAB zones. More than 60 typical harmful algae species will be isolated and their biological features including morphology, molecular, pigments, and toxicity will be recorded.

Figure 1 shows the HAATC sampling sites along the coast of China. The f ield cruise, site sampling,laboratory analysis, and data processing are combined in the research. Research vessels are used for overall distribution surveys of microalgae, cysts, and algal toxins in off shore water. Fishing boats are used in eight key zones for red tides and aquaculture, and in one key zone for green tides, where sampling will be carried out in three successive years. The annual plan of the 5-year project is to standardize the investigation protocols and calibrate the methods in the f irst year,conduct sampling and analysis from the second year to the fourth year, and then focus on the atlas and database construction in the f inal year.

Fig.1 Large-scale sampling sites (a) and key zones for successive sampling (b) along the coast of China

7 HAATC RESEARCH TEAM

Nearly 100 scientists and PhD students from the Institute of Oceanology, Chinese Academy of Sciences, Wuhan Document and Information Center, Jinan University, Xiamen University, Ocean University of China, South China Normal University,Ningbo University, Yellow Sea Fisheries Research Institute, East China Sea Fisheries Research Institute(Chinese Academy of Fishery Sciences), and the First Institute of Oceanography (Ministry of Natural Resources) participate in seven tasks:

I. Investigation of the distribution of toxic and harmful microalgae, and algal toxins in the South China Sea.

II. Investigation of the distribution of toxic and harmful microalgae, and algal toxins in the East China Sea.

III. Investigation of the distribution of toxic and harmful microalgae, and algal toxins in the Yellow Sea and Bohai Sea in China.

IV. Investigation of the distribution of toxic and harmful microalgae dormant cells in the off shore sediments of China.

V. Investigation of the species and distribution of marine harmful macroalgae in China’s coastal waters.

VI. Investigation of the morphological, genetic,pigment, and toxicity features of harmful algae species.

VII. Construction of a HAB database and information platform.A Consultation Committee is led by Prof. Yiyu CHEN (Academician), with Profs. Yuzao QI,Mingjiang ZHOU, Songhui LÜ, Rencheng YU,Song SUN, Ping XIE, Wenjun DING, Zhongjie YOU, Xiaoyong SHI, and Tian YAN from the Chinese Academy of Sciences, universities, and other institutes, as well as the Administration of Fishery and Marine Disasters.

8 SUMMARY OF LATEST HAATC PROGRESS

A list of 104 HAB species (shown in Table 1) has been proposed after several discussion meetings,determined by both morphology and genetic parameters measured in the study. Protocols for the sampling and analysis of algae (microalgae and macroalgae, and cysts), toxins (PSTs, lipid-soluble shellf ish toxins and domoic acid in phytoplankton and shellf ish samples), toxicity test methods (biology and hemolytic toxicity) for harmful algal species,and environmental data were recommended to the research teams, and generally followed national or international guidelines or references. Toxin analysiscalibration among six research groups has been carried out using phytoplankton and shellf ish samples.

Table 1 Toxic and harmful microalgae in Chinese waters (Yang LI and Songhui LÜ, unpublished data)

Table 1 Continued

Table 1 Continued

After more than 2 years of work, in November 2021,4 066 samples were obtained, and 10 169 algae data,14 630 toxin data, and 3 055 environmental data were analyzed. Furthermore, 7 731 microalgae operational taxonomic units have been obtained by molecular sequence analysis, and 24 440 toxic and harmful algae species have been annotated. The results have revealed that the distribution of HAB algae and algal toxins in China’s coastal waters has increased, of which 20 newly recorded species of harmful microalgae, seven species ofUlva, and 17 species of dinof lagellate cysts,as well as three new dinof lagellate species, have been found. Domoic acid toxins have started to become widely distributed. The investigation results of toxic algae and algal toxins causing mussel poisoning in Qinhuangdao sea areas have provided an important basis for seafood safety management, the reports of which have been submitted, and applied by national and local government departments. The f ield data of the key processes ofU.proliferagreen tides in the South Yellow Sea in 2019, 2020, and 2021 have provided a basis for the scientif ic evaluation of the national green tide mitigation, and the scientif ic and technological consultation report to the national and local governments. Over this timeframe, 67 papers have been published, including 56 papers indexed by Science Citation Index, 34 of which acknowledged HAATC as the f irst funder, as well as one monograph,nine patents, and one software copyright. A special issue of “Harmful Algae and Algal toxins in the Coastal Waters of China” is being organized for publication to the Journal of Oceanology and Limnology in November, 2022.

9 DATA AVAILABILITY STATEMENT

All data generated and/or analyzed during this study are included in this published article.

10 ACKNOWLEDGMENT

Special appreciation to Profs. Yuzao QI (Jinan University), Mingjiang ZHOU (IOCAS), Songhui LÜ(Jinan University), and Rencheng YU (IOCAS) for valuable discussion, suggestion, and other comments during the project application and implementation.Thanks also to Mr. Minjie SONG (IOCAS) for his help with editing and the f igures.