Community composition, toxigenicity, and environmental conditions during a cyanobacterial bloom occurring along 1,100 kilometers of the Murray River

A cyanobacterial bloom impacted over 1,100 km of the Murray River, Australia, and its tributaries in 2009. Physicochemical conditions in the river were optimal to support a bloom at the time. The data suggest that at least three blooms occurred concurrently in different sections of the river, with each having a different community composition and associated cyanotoxin profile. Microscopic and genetic analyses suggested the presence of potentially toxic Anabaena circinalis, Microcystis flos-aquae, and Cylindrospermopsis raciborskii at many locations. Low concentrations of saxitoxins and cylindrospermopsin were detected in Anabaena and Cylindrospermopsis populations. A multiplex quantitative PCR was used, employing novel oligonucleotide primers and fluorescent TaqMan probes, to examine bloom toxigenicity. This single reaction method identified the presence of the major cyanotoxin-producing species present in these environmental samples and also quantified the various toxin biosynthesis genes. A large number of cells present throughout the bloom were not potential toxin producers or were present in numbers below the limit of detection of the assay and therefore not an immediate health risk. Potential toxin-producing cells, possessing the cylindrospermopsin biosynthesis gene (cyrA), predominated early in the bloom, while those possessing the saxitoxin biosynthesis gene (sxtA) were more common toward its decline. In this study, the concentrations of cyanotoxins measured via enzyme-linked immunosorbent assay (ELISA) correlated positively with the respective toxin gene copy numbers, indicating that the molecular method may be used as a proxy for bloom risk assessment.     

Optical changes associated with cyanobacterial bloom termination by viral lysis

Optical changes that accompanied a collapse of the populationof filamentous cyanobacteria from a shallow, eutrophic lakewere studied in laboratory-scale enclosures (LSEs). The experimentalconditions are known, from previous work on these systems, tocause a dramatic collapse of the dominant algal or cyanobacterialspecies, which in turn can be associated with viral activity.Within 2 weeks of continuous addition of nutrient-rich growthmedium, near-complete collapse of the dominant population occurredover the span of a few days. The collapse was repeatedly andreproducibly observed and was primarily characterized by a markedincrease in water transparency. Scattering of light decreasedby 80%, absorption decreased by 20–80%. There was highsimilarity in optical changes between several experiments, carriedout in different seasons. An increase of dissolved materialand submicron-sized particles (SMP) that showed chlorophylla (Chl a) absorption was observed during the collapse. The phycocyanin(PC): Chl a ratio and phaeopigment : Chl a ratio proved to begood indicators of the observed collapse. Reflectance spectrathat were modelled using a constant volume-scattering functionindicated that mass mortality of this magnitude can be detectedin natural systems using current remote sensors.     

The role of microcystins in heavy cyanobacterial bloom formation

Regrettably, Figure 1 (part B) on p.695 was incorrect. The correctfigure appears below:     

Patchiness, collapse and succession of a cyanobacterial bloom evaluated by synoptic sampling and remote sensing

Spatial patchiness and collapse of the cyanobacterium, Nodulariaspumigena, were evaluated in saline Pyramid Lake, Nevada, bytwo synoptic ground surveys and Landsat remote sensing. Horizontalvariation of surface water chlorophyll a during the first surveyranged from 3.6 to 9790 mg m^–3 and Nodularia biovolumewas between 416?10³ and 347?10⁶ µm³ ml^–1. Differencesin spatial and temporal resolution between synoptic ground surveys({small tilde}10m, hours) and Landsat imagery (80m, seconds)yielded a poor correlation when data were matched by commonground location. A regression model for estimating chlorophyllfrom Landsat radiance was developed by pairing equivalent frequenciesfrom cumulative relative frequency distributions of both variables.Wind driven advection precipitated bloom collapse shortly afterthe first synoptic sample and Nodularia mineralization producedhigh epilimnetic ammonium concentrations. Ammonium, silica fromfluvial sources, and density differences between river and lakewater masses stimulated succession to a Chaetoceros elmoreibloom within 8 days of the first synoptic survey. The 16 dayfly by interval of Landsat is too long to document such short-termbloom succession. Landsat imagery is most applicable for evaluatinginstantaneous, basin-scale horizontal patchiness and averagelakewide chlorophyll concentrations, while frequent synopticground surveys yield more accurate estimates of meso- and micro-scalephytoplankton patchiness and species succession.     

The role of microcystins in heavy cyanobacterial bloom formation

The presence of high microcystin concentrations in cyanobacterialblooms additionally affects species diversity. Blooms with hightoxin contents can reach higher cell densities, which is alsodemonstrated by microcystin cell contents. In vitro experimentsshow that microcystins influence phytoplankton proliferation.The action is strongly dependent on the phytoplankton speciestested and light conditions. We propose that the environmentalimpact of different microcystins depends on their enzymaticinhibition activity and thus could not be measured merely onthe basis of their toxicity to vertebrate species. Their rolein heavy cyanobacterial bloom and scum formation is discussed,as well as their impact on the massive proliferation of otherspecies following toxic cyanobacterial bloom degradation.     

Photosynthesis and nitrogen fixation in a cyanobacterial bloom in the Baltic Sea

Daily integrals of photosynthesis by a cyanobacterial bloom in the Baltic Sea, during the summer of 1993, were calculated from the vertical distributions of light, temperature and the organisms in the water column and from photosynthesis/irradiance curves of picoplanktonic and diazotrophic cyanobacteria isolated from the community. The distribution of chlorophyll a in size-classes <20?µm and >20?µm was monitored over 9 days that included a deep mixing event followed by calm. Picocyanobacteria formed 70% of the cyanobacterial biomass and contributed 56% of the total primary production. Of the filamentous diazotrophs that formed the other 30%, Aphanizomenon contributed 28% and a Nodularia-containing fraction 16% of the primary production. For the whole population there was little change in standardized photosynthetic O₂ production, which remained at about 31?mmol?m^?2 before and after the mixing event. There were differences, however, between the classes of cyanobacteria: in picocyanobacteria primary production hardly changed, while in Aphanizomenon it increased by 2.6 and in Nodularia it fell below zero. Total phytoplankton photosynthesis was strongly dependent on total daily insolation with the compensation point at a photon insolation of 22.7?mol?m^?2?d^?1. Similar analyses of N₂ fixation showed much less dependence on depth distribution of light and biomass: Aphanizomenon fixed about twice as much N₂ as Nodularia their; their fixation exceeded their own N demand by about 12%. Together, these species contributed 49% of the total N demand of the phytoplankton population. Computer models based on the measured light attenuation and photosynthetic coefficients indicate that growth of the cyanobacterial population could occur only in the summer months when the critical depth of the cyanobacteria exceeds the depth of mixing.     

Dynamics of a cyanobacterial bloom in a hypereutrophic reservoir,Lake Chivero,Zimbabwe

Blooming and non-blooming periods between 2004 and 2006 in a hypereutrophic reservoir, where cyanobacterial blooms have previously been reported to be permanent, presented an opportunity to characterise factors that may favour cyanobacterial dominance. As a bloom developed in May 2004, a shift to dominance by Microcystis aeruginosa, similar to competitive exclusion, was observed. The period of M. aeruginosa dominance was characterised by the lowest Secchi depth and euphotic zone depth readings, and a decline of non-buoyant species because of competitive exclusion by M. aeruginosa, which reduced light availability in the water column. After the bloom collapsed, the euphotic zone depth increased, followed by the establishment of a Cryptomonas–Cyclotella phytoplankton assemblage. Cyanobacterial dominance within the phytoplankton assemblage was favoured by an extended stratification and was limited by nitrogen (mainly ammonium) availability. Other taxa were limited by light availability, as shown by their decline when M. aeruginosa dominated. The period of extended stratification, an increase in ammonium concentration and a decrease in nitrate concentration promoted dominance by M. aeruginosa.     

Alternative alert system for cyanobacterial bloom, using phycocyanin as a level determinant

Chlorophyll a concentration and cyanobacterial cell density are regularly employed as dual criteria for determinations of the alert level for cyanobacterial bloom. However, chlorophyll a is not confined only to the cyanobacteria, but is found universally in eukaryotic algae. Furthermore, the determination of cyanobacterial cell counts is notoriously difficult, and is unduly dependent on individual variation and trained skill. A cyanobacteria-specific parameter other than the cell count or chlorophyll a concentration is, accordingly, required in order to improve the present cyanobacterial bloom alert system. Phycocyanin has been shown to exhibit a strong correlation with a variety of bloom-related factors. This may allow for the current alert system criteria to be replaced by a three-stage alert system based on phycocyanin concentrations of 0.1, 30, and 700 microg/L. This would also be advantageous in that it would become far more simple to conduct measurements without the need for expensive equipment, thereby enabling the monitoring of entire lakes more precisely and frequently. Thus, an alert system with superior predictive ability based on high-throughput phycocyanin measurements appears feasible.     

Microbial community changes during a toxic cyanobacterial bloom in an alkaline Hungarian lake

The Carpathian Basin is a lowland plain located mainly in Hungary. Due to the nature of the bedrock, alluvial deposits, and a bowl shape, many lakes and ponds of the area are characterized by high alkalinity. In this study, we characterized temporal changes in eukaryal and bacterial community dynamics with high throughput sequencing and relate the changes to environmental conditions in Lake Velence located in Fejér county, Hungary. The sampled Lake Velence microbial populations (algal and bacterial) were analyzed to identify potential correlations with other community members and environmental parameters at six timepoints over 6 weeks in the Spring of 2012. Correlations between community members suggest a positive relationship between certain algal and bacterial populations (e.g. Chlamydomondaceae with Actinobacteria and Acidobacteria), while other correlations allude to changes in these relationships over time. During the study, high nitrogen availability may have favored non-nitrogen fixing cyanobacteria, such as the toxin-producing Microcystis aeruginosa, and the eutrophic effect may have been exacerbated by high phosphorus availability as well as the high calcium and magnesium content of the Carpathian Basin bedrock, potentially fostering exopolymer production and cell aggregation. Cyanobacterial bloom formation could have a negative environmental impact on other community members and potentially affect overall water quality as well as recreational activities. To our knowledge, this is the first prediction for relationships between photoautotrophic eukaryotes and bacteria from an alkaline, Hungarian lake.     

The possibility of using cyanobacterial bloom materials as a medium for white rot fungi

Aims: The present study was conducted to evaluate the possibility of using cyanobacterial bloom materials as a medium for white rot fungi and the capability of white rot fungi, Trichaptum abietinum 1302BG and Lopharia spadicea to biodegrade dried cyanobacterial bloom material taken from Taihu Lake. Methods and Results: The results showed T. abietinum 1302BG and L. spadicea could use the cyanobacterial bloom materials taken from Taihu Lake for growth to measure the mycelial plaque and dry‐weight mycelial pellicles of fungi. The removal rate of dried cyanobacterial bloom materials incubated with white rot fungi is approximately 100%. Conclusions: The cyanobacterial bloom material can be used as a glucose substitute in white rot fungi medium. The white rot fungi, T. abietinum 1302BG and L. spadicea, can also directly decrease the biomass of cyanobacterial bloom material taken from Taihu Lake. Significance and Impact of the Study: Cyanobacterial bloom thrives in eutrophic fresh waters all over the world. Micro‐organisms, particularly fungi, have attracted attention as possible agents for the degradation of phytoplankton species. Dealing with cyanobacterial bloom material as a medium for fungi instead of directly discharging them as organic fertilizers is a new, safe and environmentally friendly approach.     

Use of electrospray tandem mass spectrometry for identification of microcystins during a cyanobacterial bloom event

Drastic environmental conditions such as elevated temperature, abrupt pH variation, low turbulence, and high nutrient inputs can enhance the development of toxic cyanobacterial blooms in lakes and reservoirs. This study describes the occurrence of four microcystin variants (MC) in a bloom in the eutrophic reservoir Billings, in S?o Paulo City. The bloom sample was collected in October 2003, and Microcystis were the main genus found. The MC were separated and purified by reverse phase high performance liquid chromatography (RP-HPLC). Their structures were elucidated by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and four MC variants were determined: MC-RR, MC-LR, MC-YR, and MC-hRhR. MC-hRhR is described for the first time as a new variant of MC with two homoarginines at positions 2 and 4 in its structure. ESI-MS/MS analysis thus provides a powerful and convenient tool for the determination of variants of MC. These results represent an important contribution to the knowledge of the biochemistry of toxic cyanobacteria and their toxins, specifically in S?o Paulo State.     

卵孢金孢藻(Chrysosporum ovalisporum)——中国水华蓝藻新记录

卵孢金孢藻(Chrysosporum ovalisporum(Forti)Zapomelováet al.)是一种在欧洲地中海地区和澳大利亚较常见的水华蓝藻,但我国目前还未有该物种分布的报道。本文对采自上海市滴水湖中的藻种进行分离培养,获得了一株纯化藻株CFWA01007,经光学显微镜观察,初步判定其为卵孢金孢藻,对其形态特征进行了详细描述并测定了其16S rRNA基因序列,结果与来自欧洲和北美的Chrysosporum ovalisporum(Forti)Zapomelováet al.相似度极高,故判断其即为卵孢金孢藻,这是该种在中国的首次发现。对该种的分类学和生态学特征进行了讨论。     

Monitoring the colonization of monuments by bacteria: cultivation versus molecular methods

Building materials commonly used in wall paintings and monuments (mortar, limestone and sandstone) were inoculated with an artificial consortium composed of 14 microorganisms and incubated for 6 months at 28 degrees C. The colonization of the different materials by the consortium was investigated. Culture-independent techniques revealed the presence of a diversity of bacteria, whereas culture-dependent techniques yielded mainly spore-forming bacteria. The data suggest that plating leads to an overestimation of the number of spore-forming bacteria with respect to quiescent vegetative forms; the latter are less easily cultured, but are readily detected by culture-independent techniques.     

Seasonal dynamic of phototrophic epibionts on crustacean zooplankton in a eutrophic reservoir with cyanobacterial bloom

Seasonality of burden and prevalence of phototrophic (microalgal) epibionts Characidiopsis ellipsoidea, Colacium vesiculosum and Colacium sp. on dominating crustacean zooplankton (Daphnia longispina, Cyclops vicinus and Mesocyclops leuckarti) were studied in a small reservoir Bugach with cyanobacterial bloom. The correlations between the seasonal dynamics of prevalence and the dynamics of others biotic and abiotic factors were calculated. The conclusions were as follows. The substrate species, that determined the development of the epibionts on the three studied crustacean zooplankton, was Daphnia longispina (Cladocera). Despite intensive epibiotic infestation of crustacean zooplankton, epibionts did not appear to have caused non-consumptive mortality of the crustacean zooplankton. But they could have contributed to the Daphnia summer decline by increasing mortality due to its consumption by planktivorous fishes. The phototropic epibionts may successfully coexist with cyanobacterial bloom. The possible role of the epibionts in changing nutrient fluxes in pelagic food web is discussed.     

Utilization of cyanobacterial biomass from water bloom for bioproduction of lactic acid

Cyanobacterial biomass obtained from water blooms was successfully utilized as a material for lactic acid production. The starch contained in the biomass could be converted to D- and L-lactic acid with 80–90% yield by Lactobacillus amylovorus, in a manner similar to that contained in laboratory-cultured cyanobacterial biomass. The starch was also available for L-lactic acid production with similar high yields by L. agilis and L. ruminis that specifically produce L-lactic acid. The lactic acid production from the cyanobacterial biomass did not require any supplements such as yeast extract which are essential for lactic acid production from reagent soluble starch, indicating that nutrients contained in the cyanobacterial biomass might be effectively used for the production instead of the supplements. The starch content of the fresh cyanobacterial biomass from water bloom was increased from 10 to 19 and 24% by cultivation in 1 and 5% CO₂ in air, respectively. Using such starch-rich biomass, the concentration of lactic acid produced was successfully increased without changes in the conversion yield. These results indicate that wastewater bloom cyanobacteria could be utilized for the production of a useful compound, lactic acid.     

The effect of small zooplankton on the microbial loop and edible algae during a cyanobacterial bloom

SUMMARY 1. We studied the effect of the small crustacean zooplankton on heterotrophic micro-organisms and edible phytoplankton in a eutrophic lake during a cyanobacterial bloom.
2. Small (15 L) enclosures were filled with natural or screened (100 μm) lake water and incubated for 5 days in the lake. Screening removed crustacean zooplankton but the initial density of rotifers and phytoplankton remained the same in control and removal treatments. Changes in the abundance and biomass of bacteria, autotrophic picoplankton (APP), heterotrophic nanoflagellates (HNF) and ciliates were measured daily.
3. The crustacean zooplankton, dominated by the small cladoceran Chydorus sphaericus , did not affect cyanobacteria, the main phytoplankton group during the experiment.
4. The removal of the crustacean zooplankton induced a higher abundance of ciliates and reduced that of the HNF, indicating the importance of ciliates in controlling HNF in this system.     

Comparative study of various methods for determining Vibrio cholerae toxigenicity

Testing of 138 Vibrio cholerae strains for gene determinants responsible for the production of cholera enterotoxin by the polymerase chain reaction (PCR) and gene probing using molecular CT-probe showed good correlation of the results of different methods and correlation of these data with studies of V. cholerae strain virulence in vivo and in hemolytic activity test. The advantages of PCR in rapid assessment of the toxigenicity and epidemic significance of V. cholerae strains are demonstrated.     

Nitrification and denitrification by algae-attached and free-living microorganisms during a cyanobacterial bloom in Lake Taihu,a shallow Eutrophic Lake in China

Cyanobacterial blooms may stimulate epiphytic nitrification and denitrification in the water column. To validate this hypothesis, a 4-week floating mesocosms experiment that involved a cyanobacterial decay–growth–decay period was conducted at Lake Taihu. In addition to conventional methods for detecting the physical and chemical properties, quantitative real-time PCR was used to identify the nitrification and denitrification genes (archaeal and bacterial amoA, nirS and nirK). Treatment with cyanobacteria led to removal of about 3.62 mg N L^?1 total nitrogen, 40% of which was organic nitrogen, indicating a nitrogen transformation and removal mechanism was present in the system. Variations in the biogeochemical properties suggested that remineralization and coupling nitrification and denitrification by epiphytic and pelagic microorganisms was the primary pathway through which organic nitrogen was removed. The results of this study revealed that algal blooms can accelerate nitrogen removal efficiency, which may be the primary reason that nitrogen is limited in summer in Lake Taihu.     

Testing of primers for the study of cyanobacterial molecular diversity by DGGE

Denaturing Gradient Gel electrophoresis (DGGE) is a PCR-based technique which is widely used in the study of microbial communities. Here, the use of the three specific 16S rRNA cyanobacterial specific primers CYA359F, CYA781R(a) and CYA781R(b) on the assessment of the molecular diversity of cyanobacterial communities is examined. Assignments of the reverse primers CYA781R(a) and CYA781R(b) with cyanobacterial strain sequences showed that the former preferentially targets filamentous cyanobacteria whereas the latter targets unicellular cyanobacteria. The influence of the GC clamp position on the forward or on reverse primer and the use of the two reverse primers separately or in equimolar mixture were investigated. Three environmental samples were subjected to amplification with 6 combinations of primers. The 6 banding patterns as well as the sequences of the bands extracted were analysed and compared. In addition, to assess the effect of the position of the GC clamp, the melting profiles of the sequences of Aphanizomenon flos-aquae PMC9707 and Synechococcus sp. MH305 were determined, with the GC clamp in the 3' or 5' position. Results showed that the use of two separate amplifications allowed a more complete study of the molecular diversity of the cyanobacterial community investigated. Furthermore, similar richness and identical phylogenetic assignments of extracted bands were obtained irrespective of the positioning of the GC clamp.