A freshwater bacterial strain,Shewanella sp. Lzh-2, isolated from Lake Taihu and its two algicidal active substances,hexahydropyrrolo[1,2-a]pyrazine-1,4-dione and 2, 3-indolinedione

Cyanobacterial blooms have become a serious problem in Lake Taihu during the last 20 years, and Microcystis aeruginosa and Synechococcus sp. are the two dominant species in cyanobacterial blooms of Lake Taihu. A freshwater bacterial strain, Shewanella sp. Lzh-2, with strong algicidal properties against harmful cyanobacteria was isolated from Lake Taihu. Two substances with algicidal activity secreted extracellularly by Shewanella sp. Lzh-2, S-2A and S-2B, were purified from the bacterial culture of strain Lzh-2 using ethyl acetate extraction, column chromatography, and high performance liquid chromatography (HPLC) in turn. The substances S-2A and S-2B were identified as hexahydropyrrolo[1,2-a]pyrazine-1,4-dione and 2, 3-indolinedione (isatin), respectively, based on liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), and hydrogen-nuclear magnetic resonance (H-NMR) analyses, making this the first report of their algicidal activity toward cyanobacteria. S-2A (hexahydropyrrolo[1,2-a]pyrazine-1,4-dione) had no algicidal effects against Synechococcus sp. BN60, but had a high level of algicidal activity against M. aeruginosa 9110. The LD50 value of S-2A against M. aeruginosa 9110 was 5.7 μg/ml. S-2B (2, 3-indolinedione) showed a potent algicidal effect against both M. aeruginosa 9110 and Synechococcus sp. BN60, and the LD50 value of S-2B against M. aeruginosa 9110 and Synechococcus sp. BN60 was 12.5 and 34.2 μg/ml, respectively. Obvious morphological changes in M. aeruginosa 9110 and Synechococcus sp. BN60 were observed after they were exposed to S-2A (or S-2B) for 24 h. Approximately, the algicidal activity, the concentration of S-2A and S-2B, and the cell density of Lzh-2 were positively related to each other during the cocultivation process. Overall, these findings increase our knowledge about algicidal substances secreted by algicidal bacteria and indicate that strain Lzh-2 and its two algicidal substances have the potential for use as a bio-agent in controlling cyanobacterial blooms in Lake Taihu.     

Characterization of an algicidal bacterium Brevundimonas J4 and chemical defense of Synechococcus sp. BN60 against bacterium J4

As part of efforts to enhance the strategies explored to eliminate the adverse impacts of cyanobacterial blooms, we isolated an algicidal bacterium, J4, from Lake Taihu. Analysis of 16S rDNA sequence revealed that strain J4 belonged to the genus Brevundimonas. Bacterium J4 exhibited algicidal activity mainly through excretion of extracellular algicidal compounds that were further extracted with methanol and purified by silica gel chromatography and high performance liquid chromatography (HPLC). The compounds showed thermal stability, strong polarity and water solubility in J4 cultures. Study on the algicidal activity of J4 against two dominant cyanobacterial bloom-forming species in Lake Taihu showed that J4 exhibited lower algicidal rate against Synechococcus sp. BN60 (48.6%, t = 6 days) than against Microcystis aeruginosa 9110 (91.8%, t = 6 days). Additionally, rapid reduction in cell density of J4 was observed in co-cultures of Synechococcus sp. BN60 and bacterium J4 but not observed in co-cultures of M. aeruginosa 9110 and bacterium J4 during algicidal process, which was the main reason why the algicidal rate of J4 against BN60 was lower than against 9110. The reduction in cell density of J4 resulted from inducible production of antimicrobial-like compound secreted by Synechococcus sp. BN60 in co-cultures of Synechococcus sp. BN60 and bacterium J4, which reflected a kind of chemical defense from cyanobacteria (BN60) against algicidal bacteria (J4). However, M. aeruginosa 9110 had no chemical defense against J4, suggesting that whether cyanobacterial chemical defense occurs or not between cyanobacteria and algicidal bacteria depends on specific cyanobacteria–algicidal bacteria pairs. These results show that not only one-sided algicidal effect but also two-sided reciprocal inhibition interactions exist between algicidal bacteria and cyanobacteria, indicating the complexity of cyanobacteria–algicidal bacteria interactions in Lake Taihu and the need to take the cyanobacterial defensive responses into consideration when assessing potential use of algicidal bacteria.     

Molecular detection of potentially toxic cyanobacteria and their associated bacteria in lake water column and sediment

We investigated the molecular diversity of cyanobacteria and bacteria during a water bloom in a lake with a long history of toxic cyanobacterial blooms (Lake Kastoria, Greece). We also tested the hypothesis whether bloom-forming cyanobacteria are preserved in the lake’s sediment 2 years after the bloom. The dominant cyanobacteria during the bloom included the potentially toxin-producing Microcystis aeruginosa and several other Chroococcales forms closely related to the genus Microcystis. This suggests that the use of cyanobacterial-specific primers seems to be very informative in describing the cyanobacteria during the water blooms. The bacterial community showed high diversity, consisting mostly of singleton and doubleton phylotypes. The majority of the phylotypes were typical lake bacteria including some potential pathogens and toxin metabolising bacteria, suggesting that the dominant toxic cyanobacteria did not have any significant effect on the bacterial community structure. In the sediment, 2 years after the water bloom, no bloom-forming cyanobacteria were retrieved, suggesting that they cannot be preserved in the sediment. Similar to the water column, sediment bacterial diversity was also high, consisting mostly of yet-uncultured bacteria that are related to environments where organic matter degradation takes place.     

Cultivation and characterization of the MaMV-DC cyanophage that infects bloom-forming cyanobacterium Microcystis aeruginosa

The MaMV-DC cyanophage, which infects the bloom-forming cyanobacterium Microcystis aeruginosa, was isolated from Lake Dianchi, Kunming, China. Twenty-one cyanobacterial strains were used to detect the host range of MaMV-DC. Microcystic aeruginosa FACHB-524 and plaque purification were used to isolate individual cyanophages, and culturing MaMV-DC with cyanobacteria allowed us to prepare purified cyanophages for further analysis. Electron microscopy demonstrated that the negatively stained viral particles are tadpole-shaped with an icosahedral head approximately 70 nm in diameter and a contractile tail approximately 160 nm in length. Using one-step growth experiments, the latent period and burst size of MaMV-DC were estimated to be 24–48 hours and approximately 80 infectious units per cell, respectively. Restriction endonuclease digestion and agarose gel electrophoresis were performed using purified MaMV-DC genomic DNA, and the genome size was estimated to be approximately 160 kb. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed four major structural proteins. These results support the growing interest in using freshwater cyanophages to control bloom-forming cyanobacterium.     

A novel actinomycete Streptomyces aurantiogriseus with algicidal activity against the toxic cyanobacterium Microcystis aeruginosa

A novel actinomycete strain (PK1) was isolated from soil in Khon Kaen Province, Thailand, and was capable of inhibiting the cyanobacterium Microcystis aeruginosa. The isolate PK1 was identified as Streptomyces aurantiogriseus based on an analysis of biochemical and morphological characteristics and 16S rDNA sequence. The algicidal activity of PK1 against M. aeruginosa depended on the growth phase of PK1, but not on the cyanobacterial growth phase. Stationary growth phase cultures of the strain PK1 exhibited the highest anti-Microcystis activity when co-cultivated with M. aeruginosa. Complete growth inhibition was observed after 8 days of co-cultivation in liquid culture medium. The algicidal compounds were extracted from PK1 with ethyl acetate and then purified by silica gel column chromatography. These partially purified compounds demonstrated algicidal activity against M. aeruginosa, suggesting that the strain PK1 provides a potential source of extracellular compounds for the control of M. aeruginosa bloom. This is the first report of anti-cyanobacterial activity from the soil actinomycete S. aurantiogriseus.     

A marine algicidal actinomycete and its active substance against the harmful algal bloom species Phaeocystis globosa

A strain O4-6, which had pronounced algicidal effects to the harmful algal bloom causing alga Phaeocystis globosa, was isolated from mangrove sediments in the Yunxiao Mangrove National Nature Reserve, Fujian, China. Based on the 16S rRNA gene sequence and morphological characteristics, the isolate was found to be phylogenetically related to the genus Streptomyces and identified as Streptomyces malaysiensis O4-6. Heat stability, pH tolerance, molecular weight range and aqueous solubility were tested to characterize the algicidal compound secreted from O4-6. Results showed that the algicidal activity of this compound was not heat stable and not affected by pH changes. Residue extracted from the supernatant of O4-6 fermentation broth by ethyl acetate, was purified by Sephadex LH-20 column and silica gel column chromatography before further structure determination. Chemical structure of the responsible compound, named NIG355, was illustrated based on quadrupole time-of-flight mass spectrometry (Q-TOF-MS) and nuclear magnetic resonance (NMR) spectra. And this compound showed a stronger algicidal activity compared with other reported algicides. Furthermore, this article represents the first report of an algicide against P. globosa, and the compound may be potentially used as a bio-agent for controlling harmful algal blooms.     

Selective inhibition of toxic cyanobacteria by β-carboline-containing bacterium Bacillus flexus isolated from Saudi freshwaters

A bacterial strain SSZ01 isolated from a eutrophic lake in Saudi Arabia dominated by cyanobacterial blooms, showed an antialgal activity against cyanobacteria species. Based on the analysis of the 16S rDNA gene sequence, the isolated strain (SSZ01) most likely belonged to the genus Bacillus with a 99% similarity to Bacillus flexus strain EMGA5. The thin layer chromatography (TLC) analysis of the ethyl acetate extract of this bacterium revealed that this strain can produce harmine and norharmane compared to different β-carboline analog standards. Harmine and norharmane were also detected in considerable amounts in bacterial growth medium, indicating a potential excretion of these compounds into the aquatic environment. The crude extract of Bacillus flexus as well as pure materials of harmine and norharmane inhibited the growth of tested species of cyanobacteria. However, the bacterial crude extract has a higher toxicity against tested species of cyanobacteria than harmine and norharmane. In addition, harmine was more toxic to cyanobacteria than norharmane. On the other hand, neither pure compounds of harmine and norharmane nor crude bacterial extract showed any antialgal activity against tested species of green algae. The results of the present study suggest that B. flexus SSZ01 or its crude extract containing harmine and norharmane could be a candidate for the selective control of cyanobacterial blooms without affecting other algal species.     

Targeted deep sequencing reveals high diversity and variable dominance of bloom-forming cyanobacteria in eutrophic lakes

Cyanobacterial blooms in eutrophic lakes are severe environmental problems worldwide. To characterize the spatiotemporal heterogeneity of cyanobacterial blooms, a high-throughput method is necessary for the specific detection of cyanobacteria. In this study, the cyanobacterial composition of three eutrophic waters in China (Taihu Lake, Dongqian Lake, and Dongzhen Reservoir) was determined by pyrosequencing the cpcBA intergenic spacer (cpcBA-IGS) of cyanobacteria. A total of 2585 OTUs were obtained from the normalized cpcBA-IGS sequence dataset at a distance of 0.05. The 238 most abundant OTUs contained 92% of the total sequences and were classified into six cyanobacterial groups. The water samples of Taihu Lake were dominated by Microcystis, mixed Nostocales species, Synechococcus, and unclassified cyanobacteria. Besides, all the samples from Taihu Lake were clustered together in the dendrogram based on shared abundant OTUs. The cyanobacterial diversity in Dongqian Lake was dramatically decreased after sediment dredging and Synechococcus became exclusively dominant in this lake. The genus Synechococcus was also dominant in the surface water of Dongzhen Reservoir, while phylogenetically diverse cyanobacteria coexisted at a depth of 10 m in this reservoir. In summary, targeted deep sequencing based on cpcBA-IGS revealed a large diversity of bloom-forming cyanobacteria in eutrophic lakes and spatiotemporal changes in the composition of cyanobacterial communities. The genus Microcystis was the most abundant bloom-forming cyanobacteria in eutrophic lakes, while Synechococcus could be exclusively dominant under appropriate environmental conditions.     

Structure and spatial patterns of macrobenthic community in Tai Lake,a large shallow lake,China

Tai Lake (Chinese: Taihu), the third-largest freshwater lake in China, suffers from harmful cyanobacteria blooms that are caused by economic development and population growth near the lake. Several studies have focused on phytoplankton in Tai Lake after a drinking water crisis in 2007; however, these studies primarily focused on microcystin bioaccumulation and toxicity to individual species without examining the effects of microcystin on macrobenthic community diversity. In this study, we conducted a survey of the lake to examine the effects of microcystine and other pollutants on marcobenthic community diversity. A totally of forty-nine species of macroinvertebrates were found in Tai Lake. Limnodrilus hoffmeisteri and Corbicula fluminea were the most abundant species. Cluster-analysis and one-way analysis of similarity (ANOSIM) identified three significantly different macrobenthic communities among the sample sites. More specifically, sites in the eastern bays, where aquatic macrophytes were abundant, had the highest diversity of macrobenthic communities, which were dominated by Bellamya aeruginosa, Bellamya purificata, L. hoffmeisteri, and Alocinma longicornis. Sites in Zhushan Bay contained relatively diverse communities, mainly composed of L. hoffmeisteri, C. fluminea, L. claparederanus, R. sinicus, and Cythura sp. Sites in the western region, Meiliang Bay and Wuli Bay had the lowest diversity, mainly composed of L. hoffmeisteri, C. fluminea, Branchiura sowerbyi, and Rhyacodrilus sinicus. In addition, the relationships between macrobenthic metrics (Shannon–Wiener, Margalef, and Pielou) and environmental variables showed that community structure and spatial patterns of macrobenthos in Tai Lake were significantly influenced by chemical oxygen demand (COD_Cr), biochemical oxygen demand (BOD₅), lead (Pb), and microcystin-LR (L for leucine and R for arginine). Our findings provide critical information that could help managers and policymakers assess and modify ecological restoration practices.     

Host-Specificity and Dynamics in Bacterial Communities Associated with Bloom-Forming Freshwater Phytoplankton

Many freshwater phytoplankton species have the potential to form transient nuisance blooms that affect water quality and other aquatic biota. Heterotrophic bacteria can influence such blooms via nutrient regeneration but also via antagonism and other biotic interactions. We studied the composition of bacterial communities associated with three bloom-forming freshwater phytoplankton species, the diatom Aulacoseira granulata and the cyanobacteria Microcystis aeruginosa and Cylindrospermopsis raciborskii. Experimental cultures incubated with and without lake bacteria were sampled in three different growth phases and bacterial community composition was assessed by 454-Pyrosequencing of 16S rRNA gene amplicons. Betaproteobacteria were dominant in all cultures inoculated with lake bacteria, but decreased during the experiment. In contrast, Alphaproteobacteria, which made up the second most abundant class of bacteria, increased overall during the course of the experiment. Other bacterial classes responded in contrasting ways to the experimental incubations causing significantly different bacterial communities to develop in response to host phytoplankton species, growth phase and between attached and free-living fractions. Differences in bacterial community composition between cyanobacteria and diatom cultures were greater than between the two cyanobacteria. Despite the significance, major differences between phytoplankton cultures were in the proportion of the OTUs rather than in the absence or presence of specific taxa. Different phytoplankton species favoring different bacterial communities may have important consequences for the fate of organic matter in systems where these bloom forming species occur. The dynamics and development of transient blooms may also be affected as bacterial communities seem to influence phytoplankton species growth in contrasting ways.     

Growth competition between Microcystis aeruginosa and Quadrigula chodatii under controlled conditions

Cyanobacteria are the dominant bloom-forming species in Lake Taihu. Understanding the competition among algae is important to control strategies for bloom formation and outbreaks in freshwater ecosystems. In this study, we demonstrate that the cyanobacterium Microcystis aeruginosa PCC7820 and the green alga Quadrigula chodatii FACHB-1080 exhibit a strong competitive inhibitory relationship under co-culture conditions, with the latter strain inhibiting the former. Several factors influence the competitive relationship between the two species, including nutrition, temperature, and organic/inorganic compounds. Q. chodatii strongly inhibited M. aeruginosa growth through the inhibition of nitrogen utilization during co-culture. Temperature was also an influential determinant of the competition capacity between the two species under eutrophic conditions: at lower temperatures (15 °C), M. aeruginosa grew better than Q. chodatii, but the difference was not significant (p?>?0.05), whereas at higher temperatures (25 °C, 35 °C), Q. chodatii grew significantly better than M. aeruginosa (p?<?0.05). Furthermore, the Q. chodatii filtrate strongly inhibited the growth of M. aeruginosa. An analysis of the crude extracts of the algae culture filtrates from uni- and co-cultures using gas chromatography mass spectrometry (GC/MS) indicated that algal metabolites, such as dibutyl phthalate and beta-sitosterol, might play a key role in the competition between algae.     

Dynamics of cyanophage-like particles and algicidal bacteria causing Microcystis aeruginosa mortality

The dynamics of cyanophage-like particles and algicidal bacteria that infect the bloom-forming cyanobacterium Microcystis aeruginosa was followed in a hyper-eutrophic pond from September 1998 to August 1999. The densities of M. aeruginosa ranged between 4.0 × 10⁵ and 1.9 × 10⁷ cells ml⁻¹, whereas those of algicidal bacteria were between 4.0 and 5.1 × 10² plaque-forming units (PFU) ml⁻¹ and those of cyanophage-like particles were between <5.0 × 10² and 7.1 × 10³ PFU ml⁻¹. A significant relationship was found between the densities of algicidal bacteria and M. aeruginosa (r = 0.81, n = 69, P < 0.001), suggesting that the dynamics of the algicidal bacteria may regulate the abundance of M. aeruginosa. Occasional peaks of density of cyanophage-like particles were detected in October, June, and August, when sharp declines in M. aeruginosa cell densities were also observed. The densities of cyanophage-like particles became undetectable when the abundance of M. aeruginosa was low, suggesting the density-dependent infection of M. aeruginosa by cyanophage-like particles. Thus, we suggest that infections of both algicidal bacteria and cyanophage-like particles are important biological agents that decompose blooms of M. aeruginosa in freshwater environments. Received: August 31, 2000 / Accepted: December 6, 2000     

Novel L-amino acid oxidase with algicidal activity against toxic cyanobacterium Microcystis aeruginosa synthesized by a bacterium Aquimarina sp

A brownish yellow pigmented bacterial strain, designated antisso-27, was recently isolated from a water area of saltpan in Southern Taiwan. Phylogenetic analyses based on 16S rRNA gene sequences indicate that strain antisso-27 belongs the genus Aquimarina in the family Flavobacteriacea and its only closest neighbor is Aquimarina spongiae (96.6%). Based on screening for algicidal activity, strain antisso-27 exhibits potent activity against the toxic cyanobacterium Microcystis aeruginosa. Both the strain antisso-27 bacterial culture and its culture filtrate show algicidal activity against the toxic cyanobacterium, indicating that an algicidal substance is released from strain antisso-27. The algicidal activity of strain antisso-27 occurs during the late stationary phase of bacterial growth. Strain antisso-27 can synthesize an algicidal protein with a molecular mass of 190 kDa, and its isoelectric point is approximately 9.4. This study explores the nature of this algicidal protein such as l-amino acid oxidase with broad substrate specificity. The enzyme is most active with l-leucine, l-isoleucine, l-methionine and l-valine and the hydrogen peroxide generated by its catalysis mediates algicidal activity. This is the first report on an Aquimarina strain algicidal to the toxic M. aeruginosa and the algicidal activity is generated through its enzymatic activity of l-amino acid oxidase.     

Wheat Bran Enhances the Cytotoxicity of Immobilized Alcaligenes aquatilis F8 against Microcystis aeruginosa

Algicidal bacteria offer a promising option for killing cyanobacteria. Therefore, a new Alcaligenes aquatilis strain F8 was isolated to control Microcystis aeruginosa in this study. The algicidal activity of strain F8 was dependent on the cell density of M. aeruginosa, and the maximal algicidal rate of the free bacterium reached 88.45% within 72 h. With a view to its application to the control of M. aeruginosa in the natural environment, strain F8 was immobilized in sodium alginate beads, but immobilization of the strain decreased its algicidal rate compared to that of the free bacterium. However, addition of wheat bran to the sodium alginate matrix used to immobilize strain F8 not only eliminated the adverse effects of immobilization on the bacteria but also resulted in an 8.83% higher algicidal rate of the immobilized than free bacteria. Exclusion and recovery methods were used to identify key ingredients of wheat bran and gain insight into the mechanism underlying the observed enhancement of algicidal activity. This analysis indicated that certain factors in wheat bran, including vitamins B₁, B₂, B₉, and E were responsible for promoting bacterial growth and thereby improving the algicidal rate of immobilized strain F8. Our findings indicate that wheat bran is able to improve the algicidal efficiency of A. aquatilis strain F8 for killing M. aeruginosa and is a good source of not only carbon and nitrogen but also vitamins for bacteria.     

Cyanobactericidal effect of <Emphasis Type="Italic">Rhodococcus</Emphasis> sp. isolated from eutrophic lake on <Emphasis Type="Italic">Microcystis</Emphasis> sp

A bacterium, which was observed in all cultivations of Microcystis sp., was isolated and designated as Rhodococcus sp. KWR2. The growth of bloom-forming cyanobacteria, including four strains of Microcystis aeruginosa and Anabaena variabilis, was suppressed by up to 75–88% by 2% (v/v) culture broth of KWR2 after 5 days. But KWR2 did not inhibit eukaryotic algae, Chlorella vulgaris and Scenedesmus sp. An extracellular algicidal substance produced by KWR2 showed a cyanobactericidal activity of 94% and was water-soluble with a molecular weight of lower than 8 kDa.     

Isolation and Characterization of a Cyanophage Infecting the Toxic Cyanobacterium Microcystis aeruginosa

We isolated a cyanophage (Ma-LMM01) that specifically infects a toxic strain of the bloom-forming cyanobacterium Microcystis aeruginosa. Transmission electron microscopy showed that the virion is composed of anisometric head and a tail complex consisting of a central tube and a contractile sheath with helical symmetry. The morphological features and the host specificity suggest that Ma-LMM01 is a member of the cyanomyovirus group. Using semi-one-step growth experiments, the latent period and burst size were estimated to be 6 to 12 h and 50 to 120 infectious units per cell, respectively. The size of the phage genome was estimated to be ca. 160 kbp using pulse-field gel electrophoresis; the nucleic acid was sensitive to DNase I, Bal31, and all 14 restriction enzymes tested, suggesting that it is a linear double-stranded DNA having a low level of methylation. Phylogenetic analyses based on the deduced amino acid sequences of two open reading frames coding for ribonucleotide reductase alpha- and beta-subunits showed that Ma-LMM01 forms a sister group with marine and freshwater cyanobacteria and is apparently distinct from T4-like phages. Phylogenetic analysis of the deduced amino acid sequence of the putative sheath protein showed that Ma-LMM01 does not form a monophyletic group with either the T4-like phages or prophages, suggesting that Ma-LMM01 is distinct from other T4-like phages that have been described despite morphological similarity. The host-phage system which we studied is expected to contribute to our understanding of the ecology of Microcystis blooms and the genetics of cyanophages, and our results suggest the phages could be used to control toxic cyanobacterial blooms.     

Broad-spectrum In vitro antimicrobial activities of Streptomyces sp. strain BCNU 1001

Streptomyces sp. strain BCNU 1001 was isolated from forest soil samples. Cultural, morphological, and physiological characteristics as well as 16S rDNA analysis revealed that the isolate, BCNU 1001, belonged to the genus Streptomyces. The antimicrobial activity of the ethyl acetate extract was confirmed using the broth microdilution technique. The minimum inhibitory concentration (MIC) of the BCNU 1001 ethyl acetate extract was 0.25 mg/mL for Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa, and 0.125 mg/mL for Micrococcus luteus, Staphylococcus aureus, and Pseudomonas fluorescens. The MIC of the BCNU 1001 ethyl acetate extract for Aspergillus niger, Candida albicans, and Saccharomyces cerevisiae was 0.5, 0.125, and 0.25 mg/mL, respectively. BCNU 1001 was also active against dermatophytic fungi such as Trichophyton mentagrophytes and T. rubrum. Furthermore, BCNU 1001 was also found to be effective against Methicillin-resistant Staphylococcus aureus (MRSA), and its ethyl acetate extract showed MIC = 0.5 mg/mL against MRSA. The most abundant antimicrobial compound was identified as a 2-hydroxybenzyl alcohol through analysis utilizing a nuclear magnetic resonance spectroscopy. This compound was seen to be very effective against some kinds of bacteria and fungi.     

Characterization of the Biocidal Spectrum of Extracellular Filtrates of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis>

The effect of dichloromethane (DCM) and ethyl acetate (EA) extract of a cyanobacterium, Microcystis aeruginosa was evaluated against unicellular cyanobacteria and the phytopathogenic fungus Rhizoctonia solani. Fractionation of the filtrate showed the presence of five spots of different Rf values on silica gel coated plates indicating the presence of a number of compounds in the extract. A marked reduction in growth (52%) of the fungus was recorded on the plates supplemented with cyanobacterial extract, indicating the involvement of anti-fungal metabolite(s). The extract did not show any negative influence on seed germination and growth of seedlings of wheat, rice and mung, emphasizing the suitability of the compound for use in agriculture.     

Intraspecific variability in response to phosphorus depleted conditions in the cyanobacteria Microcystis aeruginosa and Raphidiopsis raciborskii

Phosphorus loading plays an important role in the occurrence of cyanobacterial blooms and understanding how this nutrient affects the physiology of cyanobacteria is imperative to manage these phenomena. Microcystis aeruginosa and Raphidiopsis raciborskii are cyanobacterial species that form potentially toxic blooms in freshwater ecosystems worldwide. Blooms comprise numerous strains with high trait variability, which can contribute to the widespread distribution of these species. Here, we explored the intraspecific variability in response to phosphorus depleted conditions (P-) testing five strains of each species. Strains could be differentiated by cell volume or genetic profiles except for those of the same species, sampling location and date, though these presented differences in their response to (P-). Although differently affected by (P-) over 10 days, all strains were able to grow and maintain photosynthetic activity. For most M. aeruginosa and R. raciborskii strains growth rates were not significantly different comparing (P+) and (P-) conditions. After ten days in (P-), only one M. aeruginosa strain and two R. raciborskii strains showed reduction in biovolume yield as compared to (P+) but in most strains chlorophyll-a concentrations were lower in (P-) than in (P+). Reduced photosystem II efficiency was found for only one R. raciborskii strain while all M. aeruginosa strains were affected. Only two M. aeruginosa and one R. raciborskii strain increased alkaline phosphatase activity under (P-) as compared to (P+). Variation in P-uptake was also observed but comparison among strains yielded homogeneous groups comprised of representatives of both species. Comparing the response of each species as a whole, the (P-) condition affected growth rate, biovolume yield and chlorophyll yield. However, these parameters revealed variation among strains of the same species to the extent that differences between M. aeruginosa and R. raciborskii were not significant. Taken together, these results do not support the idea that R. raciborskii, as a species, can withstand phosphorus limitation better than M. aeruginosa and also point that the level of intraspecific variation may preclude generalizations based on studies that use only one or few strains.     

Marine bacteria antagonistic to the harmful algal bloom species Alexandrium tamarense (Dinophyceae)

As part of efforts to enhance the strategies employed to manage and mitigate algal blooms and their adverse effects, algicidal bacteria have shown promise as potential suppressors of these events. Nine strains of bacteria algicidal against the toxic dinoflagellate, Alexandrium tamarense, were isolated from the East Sea area, China. Sequence analysis of 16S rDNA showed that all the algicidal bacteria belonged to the γ-proteobacteria subclass and the genera Pseudoalteromonas (strain SP31 and SP44), Alteromonas (strain DH12 and DH46), Idiomarina (strain SP96), Vibrio (strain DH47 and DH51) and Halomonas (strain DH74 and DH77). To assess the algicidal mode of these algicidal bacteria, bacterial cells and the filtrate from bacterial cultures were inoculated into A. tamarense cultures, and fluorescein diacetate vital stain was applied to monitor the growth of the algal cells. The results showed that all the algicidal bacteria exhibited algicidal activity through an indirect attack since algicidal activity was only detected in cell free supernatants but not the bacterial cells. This is the first report of bacteria from the genus Idiomarina showing algicidal activity to the toxic dinoflagellate A. tamarense and these findings would increase our knowledge of bacterial–algal interactions and the role of bacteria during the population dynamics of HABs.