Relationship of cellular phosphorus in the cyanobacterium Nodularia to phosphorus availability in the Peel-Harvey Estuarine System

A laboratory-determined critical concentration of phosphorus in the cyanobacterium Nodularia spumigena was used to assess the phosphorus sufficiency of field populations in the Peel-Harvey Estuarine System, Western Australia. Nutrient addition bioassays indicated that phosphorus was the limiting nutrient. Since river flow, and thus phosphorus input, to the estuary ceases 1–2 months before the onset of Nodularia blooms, the phosphorus contained in these blooms was derived from sediment release and nutrient recycling. A progressive increase in the mean cellular phosphorus concentration of Nodularia from 1981 to 1983 indicated that sediments were releasing a greater amount of phophorus each year. Although evidence suggests that the biomass of recent Nodularia blooms is now limited by physical factors, the additional phosphorus contained in each bloom will increase the supply of phosphorus to other organisms. Cellular phosphorus concentrations in Nodularia appeared to be correlated with river phosphorus loadings in the Peel Inlet but not in the Harvey Estuary.     

Factors Affecting the Germination of Akinetes of Nodularia spumigena (Cyanobacteriaceae)

Nutritional and physical factors which influence the germination of akinetes of Nodularia spumigena (Cyanobacteriaceae) were examined. Low concentrations of phosphorus (<0.9 μM) were required for germination. Nitrate had no effect, but ammonia, at concentrations of >45 μM, inhibited germination. Salinities of >20‰ were inhibitory to germination. Optimum temperatures were 22°C or greater. Germination did not take place in the dark, but only very low light intensities (0.5 microeinstein m⁻² s⁻¹) were necessary to initiate germination. Red light (620 to 665 nm) was required. More than 24 h of continuous exposure to light was necessary for any significant germination to occur. The conditions for germination corresponded with conditions in the Peel-Harvey Estuary, Western Australia, 2 to 3 weeks before large summer Nodularia blooms.     

Quantitative Real-Time PCR Detection of Toxic Nodularia Cyanobacteria in the Baltic Sea

A specific quantitative real-time PCR (qPCR) method was developed for the quantification of hepatotoxin nodularin-producing Nodularia, one of the main bloom-forming cyanobacteria in the Baltic Sea. Specific PCR primers were designed for subunit F of the nodularin synthetase gene (ndaF), which encodes the NdaF subunit of the nodularin synthetase gene complex needed for nodularin production. The qPCR method was applied to water samples (a total of 120 samples) collected from the Baltic Sea in July 2004. As few as 30 ndaF gene copies ml⁻¹ of seawater could be detected, and thus, the method was very sensitive. The ndaF gene copy numbers and nodularin concentrations were shown to correlate in the Baltic seawater, indicating the constant production of nodularin by Nodularia. This qPCR method for the ndaF gene can be used for detailed studies of Nodularia blooms and their formation. ndaF gene copies and nodularin were detected mostly in the surface water but also in deeper water layers (down to 30 m). Toxic Nodularia blooms are not only horizontally but also vertically widely distributed, and thus, the Baltic fauna is extensively exposed to nodularin.     

Benthic fauna affects recruitment from sediments of the harmful cyanobacterium Nodularia spumigena

Physical disturbance and feeding by macrofauna in the sediment can potentially affect bloom initiation of phytoplankton species that have benthic stages in their life cycle. In this experimental study, we investigated how different species of macrozoobenthos can affect the recruitment of Nodularia spumigena from the sediment to the water column. N. spumigena is a toxic, nitrogen-fixing filamentous cyanobacterium, which forms large summer blooms in the Baltic Sea. Benthic recruitment from resting stages (akinetes) and vegetative cells deposited on the seafloor have long been suspected to initiate the blooms. We found that, depending on species-specific traits, deposit-feeding macrofauna (an amphipod, Monoporeia affinis, a bivalve, Macoma balthica and an invasive polychaete, Marenzelleria cf. arctia) has the potential to either reduce or facilitate recruitment of this cyanobacterium. Shorter filament length in treatments with fauna than in the treatment without indicates feeding on or mechanical destruction of N. spumigena by the animals. Our results show the importance of an often overlooked aspect of phytoplankton bloom initiation, the role of macrozoobenthos.     

Effects of salinity and source of inocula on germination of Anabaena akinetes from a tidally influenced lake

1. Sedimentary akinetes (resting stages) may represent significant potential inocula for nuisance blooms of cyanobacteria. We studied the effects of salinity and sediment source on the germination and subsequent growth of Anabaena flos‐aquae akinetes from a shallow, tidally influenced lake. 2. Surface sediments collected from littoral and open‐water sites were used as inocula to culture A. flos‐aquae akinetes in four salinities (0.1, 2.2, 4.4 and 6.5) over 22 days. Akinete germination and development was followed by counting developmental stages every second day. 3. Filament growth, but not akinete germination, was inhibited by salinity and there were significantly fewer filaments at 6.5 than at 0.1 and 2.2. Cultures inoculated with littoral sediment had more akinetes, germlings and filaments than those inoculated with open‐water sediment. 4. Sediment is a potential source of inocula for Anabaena blooms in the lake, which potentially could develop solely from this source because germination and subsequent filament growth do not depend on the existence of an initial pelagic Anabaena population.     

Phosphatase activities in relation to phosphorus nutrition in Nodularia spumigena (Cyanobacteriaceae)

Alkaline phosphatase activities were examined in blooms of Nodularia spumigena Mertens in the Peel-Harvey estuarine system, Western Australia. Variations in phosphatase activity occurred geographically and diurnally, as well as over the duration of a bloom. These variations were well correlated with differences in the phosphorus nutritional status of the Nodularia. In the laboratory, Nodularia phosphatase production was inversely proportional to cellular phosphorus, but not to external soluble reactive phosphorus.     

Tracing the origin of green macroalgal blooms based on the large scale spatio-temporal distribution of Ulva microscopic propagules and settled mature Ulva vegetative thalli in coastal regions of the Yellow Sea,China

From 2008 to 2016, massive floating green macroalgal blooms occurred annually during the summer months in the Yellow Sea. The original source of these blooms was traced based on the spatio-temporal distribution and species composition of Ulva microscopic propagules and settled Ulva vegetative thalli monthly from December 2012 to May 2013 in the Yellow Sea. High quantities of Ulva microscopic propagules in both the water column and sediments were found in the Pyropia aquaculture area along the Jiangsu coast before a green macroalgal bloom appeared in the Yellow Sea. The abundance of Ulva microscopic propagules was significantly lower in outer areas compared to in Pyropia aquaculture areas. A molecular phylogenetic analysis suggested that Ulva prolifera microscopic propagules were the dominant microscopic propagules present during the study period. The extremely low biomass of settled Ulva vegetative thalli along the coast indicated that somatic cells of settled Ulva vegetative thalli did not provide a propagule bank for the green macroalgal blooms in the Yellow Sea. The results of this study provide further supporting evidence that the floating green macroalgal blooms originate from green macroalgae attached to Pyropia aquaculture rafts along the Jiangsu coastline of the southern Yellow Sea.     

富营养化山仔水库沉积物微囊藻复苏的受控因子

山仔水库作为福建省福州市重要的饮用水水源地之一,从2000年起每年都周期性爆发蓝藻门微囊藻属(Microcystis)水华现象,特别是在温暖的季节。对于这个富营养化水库,是否在沉积物中存在蓝藻门微囊藻的"种源"？假设山仔水库底泥中存在蓝藻门微囊藻休眠体,一定的环境条件能够促进蓝藻门微囊藻的复苏。研究于2009年12月采集水库大坝断面5根柱状沉积物,采用正交试验的方法,模拟了温度、光照、pH值、营养盐、物理扰动和浮游动物(膨大肾形虫)等环境因子对山仔水库沉积物中蓝藻门微囊藻的复苏响应。结果表明,底泥中存在着一定数量的底栖动物和硅藻、蓝藻和绿藻等微藻,从实验结束后沉积物中微囊藻数量的减少和上覆水体中微囊藻数量的增加,可以判断在适宜的环境条件下,蓝藻门微囊藻能够复苏并上浮到上覆水体中。正交实验显著性分析表明,温度是沉积物蓝藻门微囊藻复苏的重要影响因子,光照次之,上覆水体的pH值、营养盐、物理扰动和浮游动物干扰对沉积物蓝藻门微囊藻的复苏影响作用不显著,升温有利于沉积物中微囊藻的复苏。     

Proteomic profiling of the Baltic Sea cyanobacterium Nodularia spumigena strain AV1 during ammonium supplementation

The cyanobacterium Nodularia spumigena dominates the annual, toxic summer blooms in the Baltic Sea. Although Nodularia has been receiving attention due to its production of the hepatotoxin nodularin, molecular data regarding the regulation of nitrogen fixation is lacking. We have previously reported that N. spumigena strain AV1, unlike model filamentous cyanobacteria, differentiates heterocysts in the absence of detectable nitrogen fixation activity. To further analyze the uncoupling between these two linked processes, we assessed the impact of ammonium ions on the N. spumigena metabolism using a proteomic approach. Proteomic profiling was performed at three different times during ammonium supplementation using quantitative 2-dimensional gel electrophoresis followed by MS/MS analysis. Using this approach, we identified 34 proteins, 28 of which were unique proteins that changed successively in abundance during growth on ammonium. Our results indicate that N. spumigena generally exhibits lower energy production and carbon fixation in the presence of ammonium and seems to be inefficient in utilizing ammonium as an external nitrogen source. The possibility of ammonium toxicity due to PSII damage was investigated and the results are discussed. Our findings have implications in regard to the strategies considered to manage the cyanobacterial blooms in the Baltic Sea.     

Characterization of akinetes from cyanobacterial strains and lake sediment: A study of their resistance and toxic potential

Nostocalean cyanobacteria are known to proliferate abundantly in eutrophic aquatic ecosystems, and to produce several cyanotoxins, including anatoxin-a. In this study, we investigated both the resistance and toxic potential of the akinetes (resistant cells), using cyanobacterial cultures and akinetes extracted from the sediment of Lake Aydat (France) sampled in the winter and spring. Intact and lysed akinetes were differentiated using a double control based on the autofluorescence of akinetes and SYTOX-green staining. The percentage of resistant akinetes found in several different abiotic stress conditions was highly variable, depending on the species and also on the sampling season. Thus, the resistance of akinetes and their ability to germinate seems to follow a species-specific process, and akinetes can undergo physiologic changes during the sedimentary phase of the Nostocale life cycle. This study also revealed the first evidence of anatoxin-a genes in akinetes, with anaC and anaF genes detected in akinetes from all cyanobacterial producer cultures. The low number of anaC genes, almost exclusively detected using nested PCR, in the sediment at Lake Aydat suggests a limited but existent past population of toxic Nostocales in this lake. Given the key role of akinetes in the annual cycle and subsequent summer proliferation, it can be interesting to integrate the surveillance of akinetes in the management of lakes exposed to recurrent cyanobacterial blooms.     

Age, distribution and abundance of viable resting eggs of Acartia pacifica (Copepoda: Calanoida) in Xiamen Bay, China

The distribution and abundance of viable resting eggs of copepod Acartia pacifica in Xiamen Bay, China, were determined in the laboratory by the presence of nauplii hatched from the sediments. Sediment cores to a depth of 30 cm, sliced at 1.0 cm intervals, showed that most viable resting eggs of A. pacifica occurred near the sediment surface (0-5 cm), and the number of viable eggs sharply decreased with depth of the sediment, although resting eggs remained viable as deep as 23 cm. ²¹⁰Pb analyses of the sediments indicated that the maximum age of viable eggs of A. pacifica was 20.5 years and the mean egg age was 4.3 years. The egg mortality of A. pacifica in the sediment was 0.1408 year⁻¹, or 85.92% annual egg survival, calculated by regressing ln(egg density) on the age of the sediment. The horizontal distribution of viable resting eggs ranged from 2.27×10³ to 3.85×10⁵ m⁻², with a mean value of 9.49×10⁴ m⁻². Regressions between viable eggs of A. pacifica and all fine-fraction particle size classes (at 2 μm intervals) were not significant. The accumulation of viable resting eggs that can persist for an extended period of time provided evidence for the existence of an egg bank of A. pacifica in the seabed of Xiamen Bay.     

Unravelling the systematics of Nodularia (Bivalvia,Unionidae) species from eastern Russia

Conservation efforts have been hindered by data deficient conservation status assessments, especially due to taxonomic problems. This is especially true for many eastern Russian species of freshwater mussels, where distinct classification systems have complicated their delimitation and identification. Nodularia is a widespread eastern Asian freshwater mussel genus, present from Vietnam in the south to the Magadan region in eastern Russia in the north. The number of recognized species in the genus Nodularia in eastern Russia has been inflated over the last several decades due to the use of a typological species concept, the so-called 'Comparatory Method'. This method uses a single diagnostic character for species delimitation, i.e., the arc of maximal convexity of the shell's outline. Using this classification system, 10 species were recognized for far eastern Russia under the genus Nodularia Conrad, 1853, divided into three subgenera: Nodularia s. str., Amurunio and Magadaninaia. Since it is not supported by any other classification methods, the current comparatory classification is rejected by many Russian and international scientists, who only recognize a single species for that region, i.e., Nodularia douglasiae. Therefore, the aim of the present study is to clarify the taxonomy and systematics of the Nodularia genus in far eastern Russia and adjacent territories, using a multiple dataset approach that combines distribution with detailed conchological and anatomical analyses with morphometry and COI barcoding molecular techniques. All analyses performed in this study support the existence of a single Nodularia species in eastern Russia, i.e., N. douglasiae.     

Feeding interactions of the copepods Eurytemora affinis and Acartia bifilosa with the cyanobacteria Nodularia sp.

We measured ingestion and clearance rates of two Baltic Seacalanoid copepods, Eurytemora affinis and Acartia bifilosa,on toxic and non-toxic cyanobacteria Nodularia sp. using theisotope technique. Eurytemora affinis fed actively on the non-toxicstrain and moderately actively on the toxic strain, whereasA.bifilosa totally avoided feeding on both strains. This suggeststhat A.bifilosa rejected cyanobacterial filaments due to theirnutritional inadequacy or difficult manageability. The differentresponse of E.affinis to the non-toxic and toxic strains, inturn, shows that this copepod species was able to sense thepresence of the toxin in cyanobacterial filaments and thereforefed less on the toxic strain. The interaction between A.bifilosaand Nodularia sp. was further examined (with the particle countingmethod) by measuring the clearance rates of A.bifilosa on ediblegreen flagellates in the presence of cyanobacteria. The presenceor concentration of toxic Nodularia sp. did not affect grazingrates of A.bifilosa on Brachiomonas submarina. Since earlierstudies have shown that ingestion of Nodularia sp. decreasesegg production and increases mortality in E.affinis, we suggestthat the occurrence of Nodularia sp. blooms in the Baltic Seamay favour individuals of copepod species capable of selectivefeeding, such as A.bifilosa.     

Newly isolated Nodularia phage influences cyanobacterial community dynamics

Cyanophages, that is, viruses infecting cyanobacteria, are a key component driving cyanobacterial community dynamics both ecologically and evolutionarily. In addition to reducing biomass and influencing the genetic diversity of their host populations, they can also have a wider community‐level impact due to the release of nutrients by phage‐induced cell lysis. In this study, we isolated and characterized a new cyanophage, a siphophage designated as vB_NpeS‐2AV2, capable of infecting the filamentous nitrogen fixing cyanobacterium Nodularia sp. AV2 with a lytic cycle between 12 and 18 hours. The role of the phage in the ecology of its host Nodularia and competitor Synechococcus was investigated in a set of microcosm experiments. Initially, phage‐induced cell lysis decreased the number of Nodularia cells in the cultures. However, around 18%–27% of the population was resistant against the phage infection. Nitrogen was released from the Nodularia cells as a consequence of phage activity, resulting in a seven‐fold increase in Synechococcus cell density. In conclusion, the presence of the cyanophage vB_NpeS‐2AV2 altered the ecological dynamics in the cyanobacterial community and induced evolutionary changes in the Nodularia population, causing the evolution from a population dominated by susceptible cells to a population dominated by resistant ones.     

Factors regulating recruitment from the sediment to the water column in the bloom-forming cyanobacterium Gloeotrichia echinulata

1. The influence of light, temperature, sediment mixing and sediment origin (water depth) on the recruitment of the cyanobacterium Gloeotrichia echinulata was examined in the laboratory. 2. Light and temperature were the most important factors initiating germination in G. echinulata. 3. The extent of germination (recruited biovolume) was mainly regulated by temperature and sediment mixing. Furthermore, sediment mixing significantly enhanced the frequency of observed heterocysts and colonies. 4. Despite the fact that the deep and shallow sediments contained a similar number of akinete colonies, the highest recruitment occurred from shallow sediments, indicating that akinetes from shallow sediments have a higher viability than those from deeper parts of the lake. 5. Our results support the hypothesis that shallow sediments are more important than profundal sediments for the recruitment of G. echinulata to the pelagic zone.     

The dynamics of the N/P ratio and stratification in a large nitrogen-limited estuary as a result of upwelling: a tendency for offshore Nodularia blooms

The effect of upwelling on the preconditions for noxious cyanobacterial blooms in a nitrogen-limited estuary is studied with Nodularia blooms in the Gulf of Finland especially in mind. An idealised physical–chemical model is derived and integrated. The physical development in the model agrees with a classical picture of upwelling for the scales of interest. The chemical component indicates a transient minimum of dissolved inorganic nitrogen to phosphorus ratio in the middle of the gulf, coinciding with the stratification maximum resulting from physical dynamics. A general tendency for offshore cyanobacterial blooms in similar physical and biological conditions is deduced from these results.     

Nitrogen fixation by Nodularia spumigena Mertens (Cyanobacteriaceae). 1: Field studies and the contribution of blooms to the nitrogen budget of the Peel-Harvey Estuary,Western Australia

Variations in nitrogen fixation (acetylene reduction) by Nodularia spumigena blooms in the Peel-Harvey estuarine system were examined with respect to spatial (sampling station location, and depth) and temporal (seasonal and diurnal) distribution. The annual contributions of nitrogen fixation by the blooms to the nitrogen budget of the estuary were estimated to range from 309 to 713t. Contributions by nitrogen fixation were similar to the riverine inputs in the Harvey Estuary, but lower in the Peel Inlet.The Harvey Estuary had higher biomass and total fixation rates (to 0.4 nmol C₂H₂ · ml^–1 h^–1), but the heterocyst nitrogen fixation rates were greater in the Peel Inlet (to 9 × 10^–1 nmol C₂H₂ · heterocyst^–1 · h^–1). Nitrogen fixation decreased with depth in response to light, though other factors also appeared to be involved. The rates of fixation decreased concurrently with increasing bloom age, total soluble inorganic nitrogen and salinities. Maximum daily fixation rates occurred in the early morning.     

Morphology,phylogeny, growth rate and nodularin production of Nodularia spumigena from Brazil

Blooms of the toxic cyanobacterium Nodularia spumigena occur in various locations worldwide, but have not been observed in Brazil until recently. Three Nodularia strains were isolated from summer blooms in experimental shrimp production ponds of Penaeus vannamei in Rio Grande, in southern Brazil; these strains were characterized by morphology, phylogeny, growth rate and toxicity. The strains were identified as N. spumigena based on the size of vegetative cells, heterocytes and akinetes under a light microscope and based on the number of gas vesicles per μm² under a transmission electron microscope. The 16S rRNA gene sequences of the three strains showed high identity (> 99%) with N. spumigena sequences available on the NCBI database but were grouped closer in the phylogenetic tree with N. spumigena strains from Australia and USA than those from the Baltic Sea. The growth rate in batch culture varied between 0.2 and 0.6 μ?d^?1 based on cell density, optical density and chlorophyll-a content. The three strains produced the hepatotoxin nodularin (ELISA plate kit) with similar toxicity values (4.8–4.9?µg?l^?1). We conclude that the three isolated strains are N. spumigena with similar rates of growth and nodularin production. The presence of N. spumigena now represents a potential problem in aquaculture and estuarine environments in Brazil.     

The effect of salinity on the growth,toxin production,and morphology of <Emphasis Type="BoldItalic">Nodularia spumigena</Emphasis> isolated from the Gulf of Gdańsk,southern Baltic Sea

Culture experiments on the toxic Nodularia spumigena strain NSGG-1 isolated from the Gulf of Gdańsk showed a significant effect of salinity on growth and nodularin production. Growth of the NSGG-1 strain, was optimal between 7 and 18 psu, lower at 3 and 24 psu and was significantly inhibited at the extreme salinities of 0 and 35 psu. Nodularin (NOD) content of N. spumigena, estimated by the NOD/Chla ratios, correlated positively with salinity and increased from 0 to 35 psu. The NOD/Chla ratio on day 10 of growth was high, and, reached the maximum at day 30. A sudden increase in salinity from 7 to 18 and 35 psu resulted in plasmolysis of Nodularia cells. Salinity was also observed to have other effects on NSGG-1; the filaments were longest at 7 psu, while an increased number of akinetes were formed at 35 psu. The number of heterocytes was markedly reduced at the extreme salinities. This latter finding might explain why Nodularia blooms do not occur outside a certain salinity range in nitrogen-deficient waters.     

Long term nutrient loads and chlorophyll dynamics in a large temperate Australian lagoon system affected by recurring blooms of cyanobacteria

While rare globally, blooms of the toxic cyanobacteria Nodularia spumigena are a recurring problem in a few estuaries, such as the Baltic Sea and several southern Australian estuaries. Here, we document recurring Nodularia spumigena Mertens blooms in the Gippsland Lakes, S.E. Australia; a temperate lagoon system with episodic, winter-spring dominated catchment inflows. Physico-chemical conditions exerted a strong influence over bloom development, with blooms consistently occurring at surface water salinities between 9 and 20 (average?=?15), inorganic nitrogen concentrations <0.4?μM, and inorganic nitrogen to reactive phosphorus ratios <5. There was a positive correlation between average annual chlorophyll a and total phosphorus (TP) load in years when there was no Nodularia bloom, but this relationship broke down in Nodularia bloom years, even though there was a strong correlation between in-lake TP and chlorophyll a during these years; this highlights the importance of internal sources of phosphorus to bloom development. Large catchment derived nitrate and nitrite (NO_x) inputs following wildfires and floods in 2007, led to high concentrations of NO_x within the surface waters of the Gippsland Lakes through the second half of 2007 and the start of 2008. We hypothesise that these high NO_x concentrations were a key factor leading to an unprecedented Synechococcus sp. bloom that developed in the austral summer of 2007–2008, despite conditions that would otherwise favour a Nodularia bloom.