Effect of Alexandrium tamarense on three bloom-forming algae

We investigated the allelopathic properties of Alexandrium tamarense (Laboar) Balech on the growth of Prorocentrum donghaiense Lu, Chattonella marina (Subrahmanyan) Hara et Chihara and Heterosigma akashiwo (Hada) Hada in a laboratory experiment. We examined the growth of A. tamarense, C. marina, P. donghaiense and H. Akashiwo in co-cultures and the effect of filtrates from A. tamarense cultures in various growth phases, on the three harmful algal bloom (HAB)-forming algae. In co-cultures with A. tamarense, both C. marina and H. akashiwo were dramatically suppressed at high cell densities; in contrast, the growth of P. donghaiense varied in different inoculative ratios of A. tamarense and P. donghaiense. When the ratio was 1:1 (P. donghaiense: A. tamarense), growth of P. donghaiense was inhibited considerably, while the growth of P. donghaiense was almost the same as that of the control when the ratio was 9:1. The growth difference of P. donghaiense, C. marina and H. akashiwo when co-cultured with A. tamarense indicated that the allelopathic effect may be one of the important factors in algal competition and phytoplankton succession involving A. tamarense. In addition, the filtrate from A. tamarense culture had negative impacts on these three HAB algae, and such inhibition varied with different growth phases of A. tamarense in parallel with reported values of PSP toxin content in Alexandrium cells. This implied that PSP toxin was possibly involved in allelopathy of A. tamarense. However, the rapid decomposition and inactivation of PSP toxin above pH 7 weakened this possibility. Further studies on the allelochemicals responsible for the allelopathy of A. tamarense need to be carried out in future.     

Lytic Characteristics and Identification of Two Alga-lysing Bacterial Strains

Lytic Characteristics and Identification of Two Alga-lysing Bacterial StrainsBeaulieu, S. E., M. R. Sengco, and D. M. Anderson, 2005. Using clay to control harmful algal blooms: deposition and resuspension of clay/algal flocs. Harmful Algae . (4): 123-138…     

Laboratory study on the ecological impact of sophorolipid used for harmful algae elimination

We studied the role of sophorolipid in inhibiting harmful algae bloom (HAB). Different sophorolipid concentrations were tested on marine microalgae, zooplankton, fish, and bivalve (Mytilus edulis) in laboratory. The result shows that sophorolipid could inhibit the growth of algal species selectively. Among three algae species selected, Platymonas helgolandica var. tsingtaoensis was promoted with increasing sophorolipid concentration; Isochrysis galbana was inhibited seven days later in sophorolipid concentration below 40 mg/L; and Nitzschia closterium f. minutissima was inhibited obviously in only a high sophorolipid concentration over 20 mg/L. Therefore, sophorolipid in a low concentration at <20 mg/L could remove certain harmful algae species effectively without harming other non-harmful microalgae. For other animals, sophorolipid could inhibit the growth of ciliate Strombidium sp. by 50% at 20 mg/L sophorolipid concentration after 96 h. The concentration in 96-h LC₅₀ for Calanus sinicus, Neomysis awatschensis, Lateolabrax japonicus, and Paralichthys olivaceus was 15, 150, 60, and 110 mg/L, respectively. The 24 h LC₅₀ value for Artemia salina was 600 mg/L. The relative clearance rate of mussel Mytilus edulis decreased to 80%, 40%, and 20% of the control group after being exposed to 20, 50, and 100 mg/L sophorolipid for 24 h. Therefore, the toxicity for mitigation of harmful algae bloom at previously recommended concentration of 5–20 mg/L sophorolipid is low for most tested organisms in this reaserch.     

Application of rotifer Brachionus plicatilis in detecting the toxicity of harmful algae

The toxicity of seven major HAB (harmful algal bloom) species/strains, Prorocentrum donghaiense, Phaeocystis globosa, Prorocentrum micans, Alexandrium tamarense (AT-6, non-PSP producer), Alexandrium lusitanicum, Alexandrum tamarense (ATHK) and Heterosigma akashiwo were studied against rotifer Brachionus plicatilis under laboratory conditions. The results show that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6), or A. lusitanicum could maintain the individual survival and reproduction, as well as the population increase of the rotifer, but the individual reproduction would decrease when exposed to these five algae at higher densities for nine days; H. akashiwo could decrease the individual survival and reproduction, as well as population increase of the rotifer, which is similar to that of the starvation group, indicating that starvation might be its one lethal factor except for the algal toxins; A. tamarense (ATHK) has strong lethal effect on the rotifer with 48h LC₅₀ at 800 cells/mL. The experiment on ingestion ability indicated by gut pigment change shows that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6) and A. lusitanicum can be taken by the rotifers as food, but A. tamarense (ATHK) or H. akashiwo can be ingested by the rotifers. The results indicate that all the indexes of individual survival and reproduction, population increase, gut pigment change of the rotifers are good and convenient to be used to reflect the toxicities of HAB species. Therefore, rotifer is suggested as one of the toxicity testing organisms in detecting the toxicity of harmful algae. Supported by the Knowledge Innovation Program of Chinese Academy of Sciences (No. KZCX2-YW-208), National Natural Science Foundation of China (No. 40821004, U0733006) and National Basic Research Program of China (973 Program, No. 2001CB409700)     

Algicidal activities of secondary metabolites of marine macroalgal-derived endophytic fungi

Red tide occurrs frequently and causes signi?cant damage to the environment and human health. As a result, development of new effcient and environment friendly red-tide microalgae inhibitors has gained increasing attention in recent times. Algicolous endophytic fungi with unique habitats are promising sources for active agents owing to their abundant secondary metabolites and distinguished activities. In this study, the algicidal activities of 49 marine macroalgal-derived endophytic fungi against phytoplankton Alexandrium tamarense, Prorocentrum donghaiense, Heterosigma akashiwo, and Chattonella marina were examined using 96-well microplate. Four fungal strains, including Aspergillus wentii(pt-1), A. ustus(cf-42),and A. versicolor(dl-29, pt-20), exhibited potent algicidal activities. A total of 32 pure compounds isolated from these fungi were noted to possess dif ferent degrees of algicidal activities. Of those, 11 compounds comprising ?ve anthraquinones, two terpenoids, and four steroids showed high 24-h inhibition rates for the four red tide algae, with 24 h EC_(50) values ranging from 0.01 to 14.29 μg/mL. Among them, compound1(1,5-dihydroxy-3-methoxy-7-methylanthraquinone) presented the strongest activity against H. akashiwo,and could decrease its chlorophyll a(Chl a) and superoxide dismutase contents and increase the soluble protein, malondialdehyde, and peroxidase contents. These results suggested that the identi?ed anti-algal compound might inhibit the growth of red tide algae by weakening photosynthesis(reducing Chl a content),destroying cell membrane, and damaging the antioxidant system.     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

Allelopathic effects of macroalga <Emphasis Type="Italic">Corallina pilulifera</Emphasis> on the red-tide forming alga <Emphasis Type="Italic">Heterosigma akashiwo</Emphasis> under laboratory conditions

Over the past few years, harmful algal blooms (HABs), such as red tides, have been frequently observed in coastal zones worldwide. The natural allelopathic interactions among macroalgae and red tide microalgae can alter the structure and succession of aquatic ecosystems. We investigated the influence of four environmental factors (temperature, salinity, light, and pH) on the allelopathic effects of the macroalgae Corallina pilulifera on red-tide forming Heterosigma akashiwo under laboratory conditions. Each of the factors had four levels: temperature (15, 20, 25, and 30°C), salinity (10, 20, 30, and 40), light (20, 100, 200 and 400 μmol/(m²?s)), and pH (5.5, 7, 8.5, and 10. Two-factor experiments were designed for each two environmental factors, with six combination treatments (temperature-salinity, temperature-light, temperature-pH, salinity-light, salinity-pH, and light-pH). Results showed that the allelopathic effect was significantly influenced by temperature, salinity, light, and pH. As single factors, the low temperature (15°C), low salinity (10), high-intensity light (400 μmol/(m²?s)), and high pH (10) treatments substantially enhanced the allelopathic effect. The strongest allelopathic effect of C. pilulifera on H. akashiwo was observed under the following treatments: 15°C and salinity of 40, 25°C and pH 10, 25°C with medium- to high-intensity light at 200–400 μmol/(m 2 ?s), 400 μmol/(m²?s) and salinity of 10, 400 μmol/(m²?s) and pH 10, and pH 10 with a salinity of 40.     

Effect of nitrogen to phosphorus ratios on cell proliferation in marine micro algae

The ratio of nitrogen/phosphorus (N/P) is known to affect cell proliferation of some marine micro algae. We evaluated the effect of N/P ratios on the proliferation and succession of phytoplankton using five marine micro algae species. We used two sources of nitrogen, NH₄Cl (N₁) and urea (N₂), and a single source of phosphorous, NaH₂PO₄(P). The optimal N/P ratio that differed among the five species was affected by the source of nitrogen, being as follows (N₁/P, N₂/P in order): Thalassiosira sp. (30/1, 20/1), Heterosigma akashiwo (30/1, 30/1), Chroomonas salina (20/1, 30/1), Chaetoceros gracilis (40/1, 60/1), and Alexandrium sp. (10/1, 30/1). Thus, the source of nitrogen must be considered when analyzing the N/P ratio. Our results provide insight for predicting phytoplankton succession in coastal waters and may be used to forecast the potential risk of harmful algal blooms.     

Application of rotifer Brachionus plicatilis in detecting the toxicity of harmful algae

The toxicity of seven major HAB (harmful algal bloom) species/strains, Prorocentrum donghaiense, Phaeocystis globosa, Prorocentrum micans, Alexandrium tamarense (AT-6, non-PSP producer), Alexandrium lusitanicum, Alexandrum tamarense (ATHK) and Heterosigma akashiwo were studied against rotifer Brachionus plicatilis under laboratory conditions. The results show that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6), or A. lusitanicum could maintain the individual survival and reproduction, as well as the population increase of the rotifer, but the individual reproduction would decrease when exposed to these five algae at higher densities for nine days; H. akashiwo could decrease the individual survival and reproduction, as well as population increase of the rotifer, which is similar to that of the starvation group, indicating that starvation might be its one lethal factor except for the algal toxins; A. tamarense (ATHK) has strong lethal effect on the rotifer with 48h LC50 at 800 cells/mL. The experiment on ingestion ability indicated by gut pigment change shows that P.donghaiense, P. globosa, P. micans, A. tamarense (AT-6) and A. lusitanicum can be taken by the rotifers as food, but A. tamarense (ATHK) or H. akashiwo can be ingested by the rotifers. The results indicate that all the indexes of individual survival and reproduction, population increase, gut pigment change of the rotifers are good and convenient to be used to reflect the toxicities of HAB species. Therefore, rotifer is suggested as one of the toxicity testing organisms in detecting the toxicity of harmful algae.     

Experimental study on the interspecific interactions between the two bloom-forming algal species and the rotifer Brachionus plicatilis

The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms (HAB) species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×10⁴ cells mL⁻¹ of P. donghaiense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities (5, 10, 15 and 20 ×10⁴ cells mL⁻¹), and the increase rate of B. plicatilis population (r) at this algal density was 0.104 ± 0.015 rd⁻¹. Cell densities of P. donghaiense decreased due to the grazing of B. plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of the control.     

Identification of two Skeletonema costatum-like diatoms by fluorescence in situ hybridization

A harmful algae bloom (HAB) is a dense aggregation of algae in a marine or aquatic environment that can result in significant environmental problems. To forecast the occurrence of HAB, development of a rapid and precise detection method is urgently required. In this study, two Skeletonema costatum-like diatoms (SK-1 and SK-2), were identified morphologically under a light microscope, and detected using fluorescent in situ hybridization (FISH). Strain SK-1 was isolated from a frequently HAB affected area of the East China Sea, and strain SK-2 from an aquatic farm in Qingdao, China. Fluorescent DNA probes were designed that were complementary to the ITS sequence (including 5.8S rDNA) of strain SK-1. After hybridization, strong green fluorescence was observed in cells of strain SK-1 under an epifluorescence microscope; however, no such fluorescence was observed with strain SK-2, which indicates that probes hybridized only the DNA of the target strain, SK-1, in species-specific manner, and that the two strains do not belong to a same species. This finding was confirmed by ITS sequence analysis. The FISH technique used in this study was sensitive, simple, and rapid, and is a promising tool for detecting target HAB species in natural environments.     

Seasonal variation in functional phytoplankton groups in Xiangxi Bay, Three Gorges Reservoir

We describe the phytoplankton dynamics and structure in Xiangxi Bay, Three Gorges Reservoir. Samples were collected monthly in the surface waters between August 2007 and July 2008. We identified 10 principle functional groups. C-strategists and S/R-strategists with a wide range of tolerance dominated the phytoplankton assemblage. Seasonal variation was related to water column stability because of changes in hydraulic operation in October, January, and May. Functional group C (Asterionella formosa) and P (Aulacoseria granulata) dominated in August and September, whereas group Lo (Peridiniopsis niei) was the most abundant between February and April, forming a dinoflagellate bloom. Group B (Stephanodiscus hantzschii), X2 (Komma acudata), and Y (Cryptomonas erosa) were present throughout most of the year but were most abundant in late spring. A cyanobacterial bloom occurred from June to July, during which group M (Microcystis aeruginosa, M. wesenbergii) and H1 (Anabaena flos-aquae) were dominant. Green algae, characterized by group G (Eudorina sp., Pandorina sp., Pyramidomonas sp.) and J (Pediastrum spp., Coelastrum spp., Scenedesums spp.), were abundant after the bloom degraded. This sequence was corroborated by canonical correspondence analysis (CCA). The summary sequence of functional groups resulting from CCA was: C/P → Lo → H1/M/J/G. The dynamics of the phytoplankton community may be explained by the stability of water column, irradiance, water temperature, and nutrient structure.     

Temporal population dynamics of dinoflagellate Prorocentrum minimum in a semi-enclosed mariculture pond and its relationship to environmental factors and protozoan grazers

The ecological processes and interrelationships between protists, either autotrophic or heterotrophic, and environmental factors in mariculture ponds are largely unknown. This study investigated the temporal dynamics of potentially harmful dinoflagellate, Prorocentrum minimum (Pavillard) Schiller, and its relationship to physico-chemical factors and protozoan grazers over a complete cycle in a semi-enclosed shrimp-farming pond near Qingdao, Northern China. P. minimum occurred frequently in low numbers from June to August, followed by a sharp increase from the middle of August, reaching a single maximum peak value of 2.2×10⁵ cells L⁻¹ in October. Temporal variation in abundance was positively correlated with dissolved nitrogen, but showed a significant inverse relationship to abundance of the dominant ciliates, Tintinnopsis lohmanni and Askenasia stellaris. The results provide statistical evidence that the number of P. minimum increased with increasing nitrogen, and the suppression or shortening of algal bloom may be associated with protozoan grazers, such as Tintinnopsis lohmanni, in mariculture ponds.     

Antibacterial activity of extracts from <Emphasis Type="Italic">Zostera marina</Emphasis> against pathogens of <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> skin ulceration disease

The purpose of this study was to investigate the antibacterial activity of extracts from Zostera marina against the pathogens of Apostichopus japonicus skin ulceration disease. When 95% ethanol (v/v) solvent was used to extract Zostera marina at 50°C, aqueous extract (ZA) showed obvious bacteriostatic effects on the tested bacterial strains (inhibition halo diameters between 8.23 mm and 13.62 mm), whereas the ethyl acetate extract (ZE) was almost inactive. The minimal inhibitory concentration (MIC) of ZA against four pathogens were homogeneous at 12.8 g L⁻¹. ZA components were analyzed by thin layer chromatography (TLC) assay and six fractions were obtained. In another study, the six fractions showed inhibitory effects against the tested bacteria while their functions seemed to counteract the ZA activity.     

Morphology and molecular identification of Ulva forming green tides in Qingdao, China

Green tides are caused by the proliferation of chlorophytes under suitable hydrographic conditions.These blooms lead to environmental degradation and negatively impact the waters and seagrass beds,as well as fishing and other recreational activities in the bay.A comprehensive ecological understanding of the bloom dynamics,including the origin and persistence,is needed to foster management decisions.The algae in the great majority of green tide blooms usually belong to two genera of Ulvophyceae,Ulva and Enteromorpha.Ulva has been observed more often in recent years.In China,green tides occurred for the first time in the middle area of the Yellow Sea in 2007,and a large-scale algae blooming broke out in the middle and southern areas of the Yellow Sea in late May 2008.We identified them as Ulva prolifera by comparative analysis of the rDNA internal transcribed spacer 1 (ITS1),5.8S and ITS2 sequences in combination with microscopic observation.Morphological differences were found between the free-floating algae and the attached thalli.Various reproduction patterns of the free-floating algae include sexual,asexual and vegetative propagations,which played important roles in the long-term green tide persistence in China.The ITS sequences of the blooming algae were identical to those of the samples from the Lianyungang sea area but were different from the attached samples from the Qingdao sea area.The results infer that the blooms are originated from other sea areas rather than from the local attached populations.     

Salinity tolerance and genetic diversity of the dinoflagellate <Emphasis Type="Italic">Oxyrrhis marina</Emphasis>

To evaluate the relationship between salinity tolerance and genetic diversity of plankton, we collected a wild species of plankton from Taipingjiao, Qingdao. The fragment of ITS1-5.8S rDNA-ITS2 was extracted and sequenced. The results showed that the plankton belongs to Oxyrrhis marina. The salinity tolerance of O. marina ranges from 4 to 60. Seven selected groups were built up to evaluate salinity tolerance and to assess genetic diversity by RAPD. The salinity tolerance comparison revealed considerable differences among groups: the strains of O. marina in group 4 could survive under salinity from 4 to 32, while the strains selected for salinity 60 died under the salinity lower than 16. Analysis of genetic diversity of the seven groups showed that the mean genetic diversity index value was 0.28, but it was only 0.16 in selected group of 4 and was 0.24 for group 60. The result of AMOVA suggested a significantly positive relationship between the salinity tolerance and genetic diversity of O. marina (P<0.01). This study indicates that consideration of intraspecific genetic divergence in O. marina might be indispensable when using it as a model in the study of salinity tolerance of wild plankton.     

Effect of enclosure and nutrient enrichment on Microcystis blooms in Donghu Lake

1 INTRODUCTION Surface blooms of nuisance cyanobacteria are often a signal of eutrophication (Jacoby et al., 2000). They are generally favored by conditions of low nitrogen to phosphorus ratios, high total phosphorus content, high water temperature, high …     

Interspecific Competition Between Levanderina fissa(Dinophyceae) and the Other Three Microalgal Species in Co-Cultures

Growth interactions between the dinoflagellate Levanderina fissa(=Gyrodinium instriatum) and the other three algal bloom species Skeletonema costatum(Bacillariophyceae), Prorocentrum micans(Dinophyceae), and Chattonella marina(Raphidophyceae) were studied using bi-or tri-algal co-cultures under laboratory conditions. The Lotka-Volterra model was applied to estimate the interspecific competition between them. Results showed that the growth of L. fissa was significantly suppressed in both bi-algal and tri-algal cultures, and the maximum cell densities in co-cultures were only between 21% and 69% of that in mono-culture. L. fissa outcompeted P. micans; however, its effects on S. costatum and C. marina were generally less than the effects of their intraspecific competitions based on the competition coefficients. Contrarily, the inhibitory effects of S. costatum and C. marina on L. fissa were much greater. S. costatum outcompeted the other three species in all co-cultures, and its competition coefficients on the other species were more than 10000 times higher than the opposite. The growth of C. marina was suppressed by S. costatum; however, it outcompeted L. fissa and P. micans. The growth of P. micans was most strongly inhibited in co-cultures, and the maximum cell densities in co-cultures were less than 15% of that in mono-species culture. The results suggested that L. fissa had few advantages in the interspecific competition. The adaptation to low salinity and the utilization of organic phosphorus might be the important reasons for L. fissa to take the competitive predominance in brackish estuarine environments.     

Free polyamine content during algal bloom succession in the East China Sea in spring 2010

We measured the concentrations and distribution of major polyamines(spermine, putrescine and spermidine) in seawater during successive spring algal blooms in an area of frequent harmful blooms in the East China Sea. Spermine, putrescine, and spermidine concentrations were analyzed by high performance liquid chromatography and ranged from 1–64, 7–81, and 0–19 nmol/L. Spermine was present at the highest concentrations, followed by putrescine and spermidine. In late April, when a diatom bloom dominated by Skeletonema costatum dispersed, polyamine concentrations increased, presumably as a result of diatom decomposition. In early May, when a dinoflagellate bloom dominated by Prorocentrum donghaiense occurred, the polyamine concentration decreased from the level seen in late April. The abundant polyamines that decomposed and were released during the diatom bloom in late April may have promoted the growth of P. donghaiense, resulting in its dominance.     

Relationship between toxic and harmful microalgae and environmental factors in typical mariculture areas of East China Sea

Toxic and harmful algal blooms are usually more frequent in mariculture areas due to the abundant trophic conditions. To investigate the relationship between toxic and harmful microalgae and environmental factors, we set up 12 stations near three mariculture regions (Gouqi Island, Sandu Bay, and Dongshan Bay) in the East China Sea. We collected samples from all four seasons starting from May 2020 to March 2021. We identified 199 species belonging to 70 genera, of which 38 species were toxic and harmful, including 24 species of Dinophyceae, 13 species of Bacillariophyceae, and 1 species of Raphidophyceae. The species composition of toxic and harmful microalgae showed a predominance of diatoms in the summer (August), and dinoflagellates in the spring (May), autumn (November), and winter (March). The cell densities of toxic and harmful microalgae were higher in summer (with an average value of 15.34×10³ cells/L) than in other seasons, 3.53×10³ cells/L in spring, 1.82×10³ cells/L in winter, and 1.0×10³ cells/L in autumn. Pseudonitzschia pungens, Prorocentrum minimum, Paralia sulcata, and Prorocentrum micans were the dominant species and were available at all 12 stations in the three mariculture areas. We selected 10 toxic and harmful microalgal species with frequency >6 at the survey stations for the redundancy analysis (RDA), and the results show that NO ^?₃ , water temperature (WT), pH, DO, and NO ^?₂ were the main factors on distribution of toxic and harmful microalgae. We concluded that the rich nutrient conditions in the East China Sea mariculture areas increased the potential for the risk of toxic and harmful microalgal bloom outbreaks.