Dinoflagellates within the genus Karenia are well known for their potential to cause harmful algal blooms and induce detrimental ecological consequences. In this study, five Karenia species, Karenia longicanalis, Karenia papilionacea, Karenia mikimotoi, Karenia selliformis, and a new species, Karenia hui sp. nov., were isolated from Chinese coastal waters. The new species exhibits the typical characteristics of the genus Karenia, including a linear apical groove and butanoyl-oxyfucoxanthin as the major accessory pigment. It is distinguished from the other Karenia species by a wide-open sulcal intrusion onto the epicone, a conical epicone with an apical crest formed by the rim of the apical groove, and a hunchbacked hypocone. It is most closely related to Karenia cristata, with a genetic divergence of 3.16 % (22 bp out of 883 bp of LSU rDNA). Acute toxicity tests indicated that the five Karenia species from China are all toxic to marine medaka Oryzias melastigma. Karenia selliformis and K. hui were very toxic to O. melastigma, resulting in 100 % mortality within 4 h and 24 h, respectively. Further analysis by high-performance liquid chromatography revealed that four species, K. selliformis, K. longicanalis, K. papilionacea and K. mikimotoi were capable of producing Gymnodimine-A (GYM-A). The highest GYM-A content was in K. selliformis (strain HK-43), in which the value was 889 fg/cell. No GYM-A was detected in the new species K. hui, however and its toxin remains unknown. Below we provide a comprehensive report of the morphology, phylogeny, pigment composition, and toxicity profiles of Karenia species along the Chinese coast. These findings contribute new insights for monitoring of Karenia species, with important toxicological and ecological implications.

Dinoflagellates are responsible for most marine harmful algal blooms (HABs) and play vital roles in many ocean processes. More than 90% of dinoflagellates are vitamin B12 auxotrophs and that B12 availability can control dinoflagellate HABs, yet the genetic basis of B12 auxotrophy in dinoflagellates in the framework of the ecology of dinoflagellates and particularly HABs, which was the objective of this work. Here, we investigated the presence, phylogeny, and transcription of two methionine synthase genes (B12-dependent metH and B12-independent metE) via searching and assembling transcripts and genes from transcriptomic and genomic databases, cloning 38 cDNA isoforms of the two genes from 14 strains of dinoflagellates, measuring the expression at different scenarios of B12, and comprehensive phylogenetic analyses of more than 100 organisms. We found that 1) metH was present in all 58 dinoflagellates accessible and metE was present in 40 of 58 species, 2) all metE genes lacked N-terminal domains, 3) metE of dinoflagellates were phylogenetically distinct from other known metE genes, and 4) expression of metH in dinoflagellates was responsive to exogenous B12 levels while expression of metE was not responding as that of genuine metE genes. We conclude that most, hypothetically all, dinoflagellates have either non-functional metE genes lacking N-terminal domain for most species, or do not possess metE for other species, which provides the genetic basis for the widespread nature of B12 auxotrophy in dinoflagellates. The work elucidated a fundamental aspect of the nutritional ecology of dinoflagellates.

The escalating proliferation of cyanobacteria poses significant taste and odor (T/O) challenges, impacting freshwater ecosystems, public health, and water treatment costs. We examined monthly variations in four T/O compounds from September 2011 to August 2012 in Chaohu Lake\'s eastern drinking water source (DECL). More importantly, we compared the reported T/O occurrence and the related factors in freshwater bodies worldwide. The assessment of T/O issues indicated a severe and widespread problem, with many cases surpassing odor threshold values. Remarkably, China reported the highest frequency and severity of odor-related problems. A temporal analysis revealed variations in odor occurrences within the same water body across different years, emphasizing the need to consider high values in all seasons for water safety. Globally, T/O issues were widespread, demanding attention to variations within the same water body and across different layers. Algae were crucial contributors to odor compounds, necessitating targeted interventions due to diverse odorant sources and properties. A correlation analysis alone lacked definitive answers, emphasizing the essential role of further validation, such as algae isolation. Nutrients are likely to have influenced the T/O, as GSM and MIB correlated positively with nitrate and ammonia nitrogen in DECL, resulting in proposed control recommendations. This study offers recommendations for freshwater ecosystem management and serves as a foundation for future research and management strategies to address T/O challenges.

Beibu Gulf is an important semi-enclosed bay located in the northwestern South China Sea, and is famous for its high bio-productivity and rich bio-diversity. The fast development along the Beibu Gulf Economical Rim has brought pressure to the environment, and algal blooms occurred frequently in the gulf. In this study, surface water samples and micro-plankton samples (20-200 μm) were collected in the northern Beibu Gulf coast. Diversity and distribution of eukaryotic planktonic microalgae were analyzed by both metabarcoding and microscopic analyses. Metabarcoding revealed much higher diversity and species richness of microalgae than morphological observation, especially for dinoflagellates. Metabarcoding detected 144 microalgal genera in 8 phyla, while microscopy only detected 40 genera in 2 phyla. The two methods revealed different microalgal community structures. Dinoflagellates dominated in microalgal community based on metabarcoding due to their high copies of 18 s rRNA gene, and diatoms dominated under microscopy. Altogether 48 algal bloom and/or toxic species were detected in this study, 34 species by metabarcoding and 19 species by microscopy. Our result suggested a high potential risk of HABs in the Beibu Gulf. Microalgal community in the surface water samples demonstrated significantly higher OTU/species richness, alpha diversity, and abundance than those in the micro-plankton samples, although more HAB taxa were detected by microscopic observations in the micro-plankton samples. Furthermore, nano-sized taxa, such as those in chlorophytes, haptophytes, and chrysophyceans, occurred more abundantly in the surface water samples. This study provided a comprehensive morphological and molecular description of microalgal community in the northern Beibu Gulf.

Ships\' ballast water and sediments have long been linked to the global transport and expansion of invasive species and thus have become a hot research topic and administrative challenge in the past decades. The relevant concerns, however, have been mainly about the ocean-to-ocean invasion and sampling practices have been almost exclusively conducted onboard. We examined and compared the dinoflagellate cysts assemblages in 49 sediment samples collected from ballast tanks of international and domestic routes ships, washing basins associated with a ship-repair yard, Jiangyin Port (PS), and the nearby area of Yangtze River (YR) during 2017-2018. A total of 43 dinoflagellates were fully identified to species level by metabarcoding, single-cyst PCR-based sequencing, cyst germination and phylogenetic analyses, including 12 species never reported from waters of China, 14 HABs-causing, 9 toxic, and 10 not strictly marine species. Our metabarcoding and single-cyst sequencing also detected many OTUs and cysts of dinoflagellates that could not be fully identified, indicating ballast tank sediments being a risky repository of currently unrecognizable invasive species. Particularly important, 10 brackish and fresh water species of dinoflagellate cysts (such as Tyrannodinium edax) were detected from the transoceanic ships, indicating these species may function as alien species potentially invading the inland rivers and adjacent lakes if these ships conduct deballast and other practices in fresh waterbodies. Significantly higher numbers of reads and OTUs of dinoflagellates in the ballast tanks and washing basins than that in PS and YR indicate a risk of releasing cysts by ships and the associated ship-repair yards to the surrounding waters. Phylogenetic analyses revealed high intra-species genetic diversity for multiple cyst species from different ballast tanks. Our work provides novel insights into the risk of bio-invasion to fresh waters conveyed in ship\'s ballast tank sediments and washing basins of shipyards.

Paralytic shellfish toxins (PSTs) produced by marine dinoflagellates significantly impact shellfish industries worldwide. Early detection on-farm and with minimal training would allow additional time for management decisions to minimize economic losses. Here, we describe and test a standardized workflow based on the detection of sxtA4, an initial gene in the biosynthesis of PSTs. The workflow is simple and inexpensive and does not require a specialized laboratory. It consists of (1) water collection and filtration using a custom gravity sampler, (2) buffer selection for sample preservation and cell lysis for DNA, and (3) an assay based on a region of sxtA, DinoDtec lyophilized quantitative polymerase chain reaction (qPCR) assay. Water samples spiked with Alexandrium catenella showed a cell recovery of >90% when compared to light microscopy counts. The performance of the lysis method (90.3% efficient), Longmire\'s buffer, and the DinoDtec qPCR assay (tested across a range of Alexandrium species (90.7-106.9% efficiency; r2 > 0.99)) was found to be specific, sensitive, and efficient. We tested the application of this workflow weekly from May 2016 to 30th October 2017 to compare the relationship between sxtA4 copies L-1 in seawater and PSTs in mussel tissue (Mytilus galloprovincialis) on-farm and spatially (across multiple sites), effectively demonstrating an ∼2 week early warning of two A. catenella HABs (r = 0.95). Our tool provides an early, accurate, and efficient method for the identification of PST risk in shellfish aquaculture.

An unusual mass mortality event (MME) of harbour seals (Phoca vitulina) and harbour porpoises (Phocoena phocoena) occurred in Denmark and Sweden in June 2007. Prior to this incident, the region had experienced two MMEs in harbour seals caused by Phocine Distemper Virus (PDV) in 1988 and 2002. Although epidemiology and symptoms of the 2007 MME resembled PDV, none of the animals examined for PDV tested positive. Thus, it has been speculated that another - yet unknown - pathogen caused the June 2007 MME. To shed new light on the likely cause of death, we combine previously unpublished veterinary examinations of harbour seals with novel analyses of algal toxins and algal monitoring data. All harbour seals subject to pathological examination showed pneumonia, but were negative for PDV, influenza and coronavirus. Histological analyses revealed septicaemia in multiple animals, and six animals tested positive for Klebsiella pneumonia. Furthermore, we detected the algal Dinophysis toxin DTX-1b (1-115 ng g-1) in five seals subject to toxicology, representing the first time DTX-1b has been detected in marine vertebrates. However, no animals tested positive for both Klebsiella and toxins. Thus, while our relatively small sample size prevent firm conclusions on causative agents, we speculate that the unexplained MME may have been caused by a chance incidence of multiple pathogens acting in parallel in June 2007, including Dinophysis toxin and Klebsiella. Our study illustrates the complexity of wildlife MMEs and highlights the need for thorough sampling during and after MMEs, as well as additional research on and monitoring of DTX-1b and other algal toxins in the region.

The role of microorganisms in effectively terminating harmful algal blooms (HABs) is crucial for maintaining environmental stability. Recent studies have placed increased emphasis on bio-agents capable of inhibiting HABs. The bacterium Pseudoalteromonas sp. strain FDHY-MZ2 has exhibited impressive algicidal abilities against Karenia mikimotoi, a notorious global HAB-forming species. To augment this capability, cultures were progressively scaled from shake flask conditions to small-scale (5 L) and pilot-scale (50 L) fermentation. By employing a specifically tailored culture medium (2216E basal medium with 1.5% soluble starch and 0.5% peptone), under precise conditions (66 h, 20 °C, 450 rpm, 30 L/min ventilation, 3% seeding, and constant starch flow), a notable increase in algicidal bacterial biomass was observed; the bacterial dosage required to entirely wipe out K. mikimotoi within a day decreased from 1% to 0.025%. Compared to an unoptimized shake flask group, the optimized fermentation culture caused significant reductions in algal chlorophyll and protein levels (21.85% and 78.3%, respectively). Co-culturing induced increases in algal malondialdehyde and H2O2 by 5.98 and 5.38 times, respectively, leading to further disruption of algal photosynthesis. This study underscores the unexplored potential of systematically utilized microbial agents in mitigating HABs, providing a pathway for their wider application.

This investigation was undertaken to characterize health effects associated with a major bloom of blue-green algae due to the proliferation Microcystis aeruginosa that occurred in Florida in 2018. Cyanobacteria produce multiple toxins, including the potent hepatotoxic microcystins (MCs), that have been reported to cause illness in exposed persons worldwide. Widespread exposure to toxins released by blue-green algae during the 2018 bloom was shown by the presence of MCs in the nasal passages of 95 percent of the individuals studied previously in south Florida (Schaefer et al., 2020). The current analyses were conducted to determine whether self-reported symptoms were associated with activity patterns, direct contact with water, residential, recreational, and occupational exposure. The 125 persons who participated in the initial study reported an average of 4.94 (± 4.87) symptoms. Those reported most commonly included rhinorrhea, sneezing, headache, sore throat and dry cough. Respiratory symptoms were reported by 74%, ocular symptoms by 62%, and gastrointestinal symptoms by 35% of respondents. Residential and recreational exposures were associated with increased risks of respiratory, gastrointestinal, or ocular symptoms in univariate and adjusted multivariable analyses. Residential exposure was significantly associated with increased reporting of dry cough (p = 0.03), dyspnea (p < 0.01) and wheezy respirations (p = 0.04). Among persons reporting gastrointestinal symptoms, nausea (p = 0.02) and abdominal pain (p < 0.01) were significantly associated with residential exposure. Recreational exposure was significantly associated with sore throat and eye irritation. The findings add to the evidence that exposure to cyanobacteria at concentrations encountered during an algal bloom is associated with a diverse array of symptoms and that inhalation of aerosols constitutes an important exposure pathway.

During the spring and summer of 2019, an unprecedented cyanobacterial harmful algal bloom (cyanoHAB) was responsible for beach advisories on 25 beaches along the Mississippi Sound for over 3 months. Due to the preceding heavy rainfall and flooding within the Mississippi River watershed, for the first time in history, the Bonnet Carré Spillway (BCS) opened twice in one year during 2019. The coastal cyanoHAB coincided with the second BCS opening. The main objectives of this study were: (1) to investigate the potential for using the National Aeronautics and Space Administration (NASA) ocean color standard Cyanobacteria Index (CIcyano) algorithm to characterize the spatial and temporal extent of the 2019 cyanoHAB; (2) to couple the CIcyano data with river discharge, salinity, and modeled-wind data to study the conditions leading to the cyanoHAB and factors aiding the advection and persistence of the bloom within the Mississippi Sound, including a possible relationship to the BCS; (3) to further investigate the relationship with the BCS by repeating the methods using data from 2018, which was a year when the BCS was opened but no evidence of cyanoHABs was reported along the Mississippi coast. Weekly means and monthly frequency CIcyano images, river discharge, salinity, and modeled-wind data from February to September of 2018 and 2019 were analyzed, which coincide with three BCS openings. In March 2018, a cyanobacteria bloom was observed within Lake Pontchartrain coinciding with the BCS opening; however, the month-long bloom was contained to the lake. Two distinct cyanoHABs were observed in 2019 and both blooms were advected into the Mississippi Sound, and likely contributed to the 3-month-long beach water advisories of 2019 along the Mississippi coastline. From March to mid-July 2019, salinity at stations within the Mississippi Sound was consistently near zero indicating high levels of freshwater. During that time, winds were predominantly northwestward, preventing the BCS waters from flushing into the Mississippi Shelf and resulting in BCS waters remaining longer within the estuarine lakes and Mississippi Sound. Although the BCS had an undeniable impact on the presence of the coastal cyanoHAB of 2019, other variables including wind direction, water flow, mixing, and persistence of freshwater within the Sound can determine the intensity and extent of the cyanoHABs. Coupling in situ phytoplankton information from freshwater water bodies to the marine continuum along with water flow, wind data, and satellite imagery could help identify cyanoHABs at early stages and forecast their trajectory and potential impacts on coastal areas.