Toxicity to Daphnia of a compound extracted from laboratory and natural Microcystis spp., and the role of microcystins

1. The microcystin content of a variety of Microcystis spp., from both laboratory strains and natural blooms, was analysed by HPLC. The microcystin content of laboratory strains ranged from 1.6 to 4.3μgmg^?1 dry weight. Yearly and seasonal variation was detected in an analysis of bloom material collected from Bautzen Reservoir over a 3-year period. The microcystin concentration in bloom material ranged from undetectable to 1.16 μg ml^?1 dry weight. 2. Toxicity of laboratory and natural Microcystis to Daphnia pulicaria was determined using an established LC₅₀ technique. Partially purified water extracts from different Microcystis samples exhibited a wide range of toxicity. The highest activity was found in natural Microcystis samples, with an LC₅₀ of 36 μgm^?1 dry weight of Microcystis, whereas one strain did not appear toxic at 1600 μg ml^?1. 3. No correlation was found between the concentrations of microcystins of different laboratory and natural Microcystis strains and the toxicity of extracts to Daphnia pulicaria from the same strains. Therefore, we discriminated between hepatotoxic microcystins and the compound(s) that is toxic to Daphnia, here termed DTC (Daphnia-toxic compound), which is independent of microcystins.     

Incorporation of [15N]Ammonia by the Cellulolytic Ruminal Bacteria Fibrobacter succinogenes BL2, Ruminococcus albus SY3, and Ruminococcus flavefaciens 17

The origin of cell nitrogen and amino acid nitrogen during growth of ruminal cellulolytic bacteria in different growth media was investigated by using ¹⁵NH₃. At high concentrations of peptides (Trypticase, 10 g/liter) and amino acids (15.5 g/liter), significant amounts of cell nitrogen of Fibrobacter succinogenes BL2 (51%), Ruminococcus flavefaciens 17 (43%), and Ruminococcus albus SY3 (46%) were derived from non-NH₃-N. With peptides at 1 g/liter, a mean of 80% of cell nitrogen was from NH₃. More cell nitrogen was formed from NH₃ during growth on cellobiose compared with growth on cellulose in all media. Phenylalanine was essential for F. succinogenes, and its ¹⁵N enrichment declined more than that of other amino acids in all species when amino acids were added to the medium.     

IMPROVED PROCEDURES FOR THE CLONING AND PURIFICATION OF MICROCYSTIS CULTURES (CYANOPHYTA)1

A cloned axenic culture of Microcystis Kützing was obtained by combining two procedures: a) the disaggregation of multicellular Microcystis colonies by dilution into deionized water, and b) the selective growth of Microcystis in agar media containing Na₂S, which inhibited or killed the associated contaminants. Microcystis growth was stimulated by 0.3–1 mM Na₂SO₃, but not by 0.1–33 mM Na₂SO₄. Although Microcystis cells survived temporary exposure to high Na₂S concentrations, their growth was not stimulated by 1 × 10^?5 to 1.0 M Na₂S. Possible metabolic roles of reduced sulfur compounds are considered. Microcystis colonies disaggregated to unicells at ionic concentrations below 1 mM for univalent cations, 10–100 μM for the divalent cations, and 3–10 μM for Fe³⁺. Higher cation concentrations prompted cell aggregation. With > 100 mM Fe³⁺, the Microcystis capsule appeared rust-colored. Neither nonionic solutes nor anions detectably influenced aggregation. These observations suggest cation interactions with the Microcystis capsule and are discussed with regard to: a) possible siderochrome activity, cation chelation or luxury uptake of cations, b) the questionability of using cell aggregation as a criterion for identifying Microcystis in samples of unknown ionic strength, c) the utility of low ionic strength media in releasing contaminating bacteria from the capsule and in obtaining algal unicells for cloning, and d) a model for cation interactions with the capsule.     

Consortium of Higher Aquatic Plants and Microalgae Designed to Purify Sewage of Heavy Metal Ions

We selected higher aquatic plants (HAP) and microalgae possessing a high sorption capacity in respect to heavy metals to form a consortium designed to purify contaminated aquatic ecosystems. Accumulation of heavy metals Cd²⁺, Cu²⁺, Pb²⁺, and Zn²⁺ was investigated in plants Pistia stratiotes, Elodea canadensis, and Lemna minor and green microalgae Chlorella vulgaris ВВ-2, Ankistrodesmus sp. ВI-1, Chlamydomonas reinhardtii В-4, and Scеnеdеsmus quadricauda В-1. It was found that intense accumulation of metals occurs in cultures of HAP Pistia stratiotes and Elodea canadensis. These plants are macroconcentrators of zinc, lead, and copper and microconcentrators of cadmium. Out of the examined cultures of microalgae, effective bioaccumulators of heavy metals were C. vulgaris ВВ-2 and Ankistrodesmus sp. ВI-1. It was shown that heavy metals are selectively taken up from the medium in the series Zn²⁺ > Cu²⁺ > Cd²⁺ > Pb²⁺. In order to produce a consortium of higher aquatic plants and microalgae for purification of polluted aquatic ecosystems, we investigated interaction of HAP P. stratiotes and E. canadensis with microalgae C. vulgaris ВВ-2 and Ankistrodesmus sp. ВI-1 in the course of their cocultivation. Neutral relations were detected between the cells of microalgae C. vulgaris ВВ-2 and Ankistrodesmus sp. ВI-1 and HAP E. canadensis. At the same time, the cells of Ankistrodesmus sp. ВI-1 and HAP P. stratiotes formed a symbiosis. Microscopic examination showed numerous points where the cells of microalgae Ankistrodesmus sp. ВI-1 were attached to the roots of P. stratiotes plants. We tested an opportunity to employ the association between P. stratiotes and Ankistrodesmus sp. ВI-1 for purification of simulated wastewater polluted with heavy metal ions. This consortium proved to be capable of eliminating contaminants from the sewage, reducing their level in the sewage to standard values, and active accumulation of heavy metal ions.     

Inhibitory and Toxic Effects of Blue-green Algae on Daphnia

The effects of the planktonic blue-green algae, Aphanizomenon gracile, Synechococcus elongatus, and Microcystis aeruginosa, on survival, growth, and food uptake of Daphnia pulicaria were determined. Synechococcus and Aphanizomenon were unsuitable food when offered alone, but did not affect the daphnids negatively when mixed with Scenedesmus. Microcystis was the only one found to be toxic. In pure suspensions of this blue-green, the daphnids did not survive more than 48 hours; they lived a little longer if Scenedesmus was supplied additionally. Growth was markedly reduced when only 50 μg carbon/l of Microcystis was added to the normal Scenedesmus food. It ceased at a concentration of 250 μg C/l. This can be explained by the reduction of food uptake. Very small quantities of Microcystis (10 μg C/l) present in the normal food caused a significant reduction of the filtering rate. Filtering inhibition was associated with the cells. Filtrate of Microcystis suspensions was not effective. Thus, the daphnids must ingest the blue-green cells in order to become toxified. Dual-labelling experiments showed that Microcystis cells are filtered from the medium by Daphnia with the same efficiency as Scendesmus and are not rejected. Toxicity of Microcystis is considered to be an effective defence mechanism against grazing pressure.     

Survival of Chlorophyceae Ingested by Saprozoic Nematodes

The saprozoic nematode, Pristionchus lheritieri ingested cells of four species of unicellular Chlorophyceae (grass-green algae) including Chlamydomonas reinhardi and unidentified species of Ankistrodesmus, Chlamydornonas and Scenedesmus. Additional tests with Ankistrodesmus sp. and Chlamydomonas sp., indicated cells of Ankistrodesmus survived passage through the alimentary canal and were subsequently cultured, while viable cells of Chlarnydomonas were only occasionally recovered.     

Urea dynamics during Lake Taihu cyanobacterial blooms in China

Lake Taihu, the third largest freshwater lake in China, suffers from harmful cyanobacteria blooms caused by Microcystis spp., which do not fix nitrogen (N). Reduced N (i.e., NH₄⁺, urea and other labile organic N compounds) is an important factor affecting the growth of Microcystis. As the world use of urea as fertilizer has escalated during the past decades, an understanding of how urea cycling relates to blooms of Microcystis is critical to predicting, controlling and alleviating the problem. In this study, the cycling rates of urea-N in Lake Taihu ranged from non-detectable to 1.37 μmol N L⁻¹ h⁻¹ for regeneration, and from 0.042 μmol N L⁻¹ h⁻¹ to 2.27 μmol N L⁻¹ h⁻¹ for potential urea-N removal. The fate of urea-N differed between light and dark incubations. Increased ¹⁵NH₄⁺ accumulated and higher quantities of the removed urea-¹⁵N remained in the ¹⁵NH₄⁺ form were detected in the dark than in the light. A follow-up incubation experiment with ¹⁵N-urea confirmed that Microcystis can grow on urea but its effects on urea dynamics were minor, indicating that Microcystis was not the major factor causing the observed fates of urea under different light conditions in Lake Taihu. Bacterial community composition and predicted functional gene data suggested that heterotrophic bacteria metabolized urea, even though Microcystis spp. was the dominant bloom organism.     

Detection of Toxigenicity by a Probe for the Microcystin Synthetase A Gene (mcyA) of the Cyanobacterial Genus Microcystis: Comparison of Toxicities with 16S rRNA and Phycocyanin Operon (Phycocyanin Intergenic Spacer) Phylogenies

The relationship between toxigenicity and phylogeny within the cyanobacterial genus Microcystis is unclear. To investigate this issue, we have designed PCR primers for the N-methyltransferase (NMT) domain of the microcystin synthetase gene mcyA and have probed 37 Microcystis sp. cultures as well as several field samples. The NMT region was present in all 18 laboratory strains that gave positive reactions in the protein phosphatase inhibition assay for microcystin but was absent in 17 nontoxic strains. Two other nontoxic strains, one of which had previously been reported to produce microcystin, possessed the NMT region. Detection of NMT-specific DNA in field samples corresponded to periods of toxicity as assessed by protein phosphatase inhibition. The Microcystis strains formed a monophyletic cluster based on 16S rRNA gene sequences but comprised two groups with respect to phycocyanin intergenic spacer (PC-IGS) sequences. Toxic and nontoxic strains appeared to be erratically distributed within the PC-IGS and 16S rRNA trees. Sequence analysis of the NMT domain revealed two coherent groups. The genomic region immediately downstream of the mcyABC cluster in all 20 NMT-positive strains contained an open reading frame of unknown function (uma1) at a conserved distance from mcyC. All nontoxic strains also contained uma1, which is not cotranscribed with mcyABC. The consistent linkage of mcyC to uma1 suggests that mcyC has not been frequently transferred into nontoxic strains via any mechanism involving insertion at random chromosomal locations. These results are discussed with respect to various mechanisms that could explain the patchy distribution of toxigenicity among the various Microcystis clades.     

Effects of toxic and non-toxic cyanobacteria on the life history of tropical and temperate cladocerans

1. This study compares the effects of four toxic strains of Microcystis aeruginosa on tropical and temperate Cladocera. Survival was tested in acute toxicity experiments using Microcystis alone or in mixtures with the edible green algae Ankistrodesmus falcatus. The effect of chronic exposure on population growth was estimated in life‐table experiments by varying the proportion of Microcystis and the green alga. Nutritional deficiency was assessed using a non‐toxic cyanobacterium in a zooplankton growth experiment. Feeding inhibition was tested using a C‐labelled green alga as a tracer in mixtures with toxic Microcystis.
2. Toxicity varied consistently between Microcystis strains, while sensitivity varied consistently between cladoceran species. However, no relationship was found between sensitivity and geographical origin or cladoceran body size. Two small‐bodied cladocerans from the same tropical lake, Ceriodaphnia cornuta and Moinodaphnia macleayi, were the least sensitive and most sensitive species, respectively.
3. Surprisingly, two small tropical cladocerans survived longer without food than did three large Daphnia species and a third small tropical species.
4. Each of the three tropical Microcystis strains strongly reduced the population growth rate (little ‘r’) and reproductive output of each cladoceran, this reduction being proportional to the percentage of toxic cells in the diet.
5. As the sole food source, the non‐toxic cyanobacterium, Synechococcus elongatus, supported poor growth in M. macleayi. The nutritional deficiency was overcome when Synechococcus was mixed with either Ankistrodesmus or an emulsion rich in omega‐3 fatty acids.
6. Microcystis inhibited the feeding rate of two cladocerans, even when it comprised only 5% of a mixture with the green algae A. falcatus.
7. Differences in sensitivity to the toxic cyanobacterium appear to be associated with differences in life history between the cladoceran species rather than differences between tropical and temperate taxa. Slow‐growing species that are resistant to starvation appear less sensitive to toxic Microcystis than fast‐growing species, which also tend to die more quickly in the absence of food.     

Algal bioassays to determine toxicity of metal mixtures

The shortcomings of many established toxicity criteria for metals have resulted from a simplification of bioassays performed with single metals. A more comprehensive approach was needed to diagnose the effects of metal mixtures on aquatic organisms. Using Chlorella, Ankistrodesmus and Scenedesmus as test organisms, we experimented on a number of factors which affected metal toxicity bioassays. These included metal interactions, algal competitions, species sensitivities, the ratio of an excess metal to other metals and nutrient levels. An alternative technique was finally established which involved an evaluation of individual assays of Chlorella and Ankistrodesmus in separate media and a tissue-metal analysis on Chlorella. Chlorella fusca, commonly found in lakes with high metal concentrations, showed high tolerance to mixed-metal solutions, while Ankistrodesmus proved to be very sensitive. By determining the maximum yield ratio between Ankistrodesmus and Chlorella (i.e. the A/Ch ratio) it was possible to compare the toxic strength of harmful metals according to an established standard curve of A/Ch ratio versus mixed-metal concentrations. The levels of tissue-metal analysed in Chlorella also gave some indication as to which metals were responsible for the toxicity.     

Toxin production of cyanobacteria is increased by exposure to zooplankton

1. Cyanobacterial toxin production in response to direct and indirect zooplankton feeding activity was examined using four strains of Microcystis aeruginosa, of which three were previously reported to be toxic to zooplankton and one non‐toxic. Direct (Microcystis cultured with zooplankton) and indirect effects (Microcystis cultured with filtered zooplankton culture media, ZCMF) were tested for the zooplankton species, Moina macrocopa, Daphnia magna or D. pulex. 2. With direct exposure to zooplankton, increased mass‐specific microcystin productions occurred in all Microcystis strains, with mean microcystin concentrations up to five times greater (61.5–177.3 μg g^?1 dry cell) than the controls. 3. With indirect exposure, mass‐specific microcystin production increased over controls in three strains of M. aeruginosa. Mean maximum concentrations of microcystin during the experiment were 92.6–125.7 μg g^?1 dry cell. 4. These results suggest that several strains of Microcystis aeruginosa increased toxin production in response to direct and indirect exposure to herbivorous zooplankton of several species, and support the hypothesis that this response is an induced defence mediated by the release of info‐chemicals from zooplankton.     

Grazing on toxic cyanobacterial blooms by tadpoles of edible frog Rana grylio

Tadpoles of Rana grylio were raised as edible frogs in fishponds of Guanqiao in Wuhan City, Hubei, China, during cyanobacterial blooms from June to October. The dominant cyanobacterial species was Microcystis, which was found to be lethally toxic by intraperitoneal (i.p.) mouse bioassay. Little is known about the effect of tadpoles on toxic cyanobacterial blooms. To evaluate the potential of the tadpoles to graze on cyanobacterial blooms, the tadpoles were fed on Microcystis collected from the field in the laboratory. The Microcystis cells decreased from 1.19 × 10⁷ cells mL^?1 to 3.23 × 10⁶ cells mL^?1, with a sharp reduction of 73% of the initial Microcystis population observed in the first 24 h after introduction of the tadpoles. The ponds containing tadpoles had a markedly lower density of Microcystis than those lacking tadpoles. Tadpoles exposed to either cultured Microcystis aeruginosa (NIES–90, 2.768 µg microcystins mg^–1 dw^–1) cells or lysed M. aeruginosa cells grew well, however, indicating that they were unaffected by Microcystis toxins. We found a significant increase in tadpole body weight after feeding on either field Microcystis or cultured M. aeruginosa. The mean increase in individual body weight was 20 mg day^?1 when fed on Microcystis from the pond, and 7 mg day^?1 when fed on M. aeruginosa from culture. Our study strongly suggested that there is a direct trophic relationship between R. grylio tadpoles and toxic Microcystis blooms and they possess the potential to graze on toxic Microcystis. The results imply that R. grylio tadpoles may play an important ecological role in reducing toxic cyanobacterial blooms caused by Microcystis.     

Physiologische und biochemische Beiträge zur Taxonomie der Gattungen Ankistrodesmus und Scenedesmus

The base composition of the DNA was found to be a species-specific, taxonomically valuable character also in the genera Ankistrodesmus (21 strains) and Scenedesmus (27 strains). As compared to the genus Chlorella, however, the differences between different species are considerably smaller. The GC content is between 63 and 70% in the genus Ankistrodesmus, and between 52 and 62% in the genus Scenedesmus. Obviously, a few strains with a very much different base composition do not belong to the genus Ankistrodesmus.     

Microbiological spoilage of mayonnaise and salad dressings

Saccharomyces bailii was isolated from two-thirds of the spoiled mayonnaise and salad dressing samples examined. Most of the rest were spoiled by Lactobacillus fructivorans. However, one sample contained large numbers of both S. bailii and L. plantarum. Two of the spoiled samples also contained small numbers of bacilli. Bacillus subtilis, B. pumilis, B. polymyxa, B. megaterium, and B. licheniformis were found in one sample and B. subtilis and B. pumilis in another. Small numbers of B. subtilis and B. licheniformis were also present in one unspoiled sample. Several media were evaluated for the isolation of L. fructivorans. S. bailii and L. fructivorans vigorously fermented glucose. The concentration of glucose in the spoiled samples ranged from 0 to 38.5 g/kg and from 1.3 to 17.8 g/kg for the unspoiled samples.     

Production of Candida utilis protein from peat extracts

Sphagnum peat extracts or hydrolysates have been obtained and used as a culture medium for the production of Candida utilis biomass as single cell proteins. Acid hydrolysis of ground peat (4–60 mesh) in an autoclave operated under a set of conditions for acid strength (0.3-1.5 (v/v) H₂SO₄), holding time (1–4 hr), temperature (100–165°C), and weight ratio of dry peat to solution (3.3–16.7 g dry peat/100 g solution) yielded carbohydrate-rich extracts of different concentrations (1–34g/liter). The best yield (mg total carbohydrate/g dry peat) was obtained for a holding time of I hr and a temperature of 152°C. Low peat concentratio (4.1 g dry peat/100 g solution)resulted in high yield(280mg total carbohydrate/gdry peat) with a corresponding low carbohydrate content in hydrolysate (13 g/liter), while a lower yield with a higher carbohydrate content (34 g/liter)in hydrolysate were found when increasing peat concentration (16.7 g dry peat/100 g solution). Shake-fladk experiments using peat hydrolysates as the culture medium together with NH₄OH (～4.8 g/liter) and K₂HPO₄(5 g/liter) as nitrogen and phosphate supplement, respectively, gave a maximum biomass concentration of 7.5 g/liter after 60 hr at 30°C and 200rpm. Batch cultivation in a fermentor under controlled conditions for aeration (4.2 liter/min), agitation (500rpm), temperature (30°C), and pH (5.0) produced a maximum biomass of 10 g/liter after 20 hr with a specific growth rate of 0.13 hr^?1. For the continuous cultivation, a maximal biomass productivity of 1.24 g/gliter-he was obtained at a dilution rate of 0.125 hr ^?1. Monod's equation's equation has been used for the estimation of the coefficients μ_Max, K_s, and Y. It was found that the yield coefficient Y is not constant during the progress of batch cultivation.     

A Field Study to Investigate Environmental Factors that Could Effect Microcystin Synthesis of a Microcystis Population in the Bautzen Reservoir

Toxicity of Microcystis blooms to warm-blooded animals generated by microcystins has bew reported world wide. The ecological relevance of microcystin production for cyanobacteria remains unknown. The microcystin concentration in Microcystis blooms occurring in the Bautzen reservoir Was investigated. The microcystin content of samples were determined by HPLC and ranged from undetectabel to 14.7 μg mg⁻¹. Various chemical and physical parameters were monitored at the same time as Microcystis sampling, however, there was no correlation between these parameters and microcystin dynamics. The spatial distribution of microcystin in the Microcystis population was investigated once and showed no difference between samples taken at five stations. The microcystin concentration in ihc cell free water from the reservoir was below the detection threshold (< 1 μg L⁻¹). Size dependent Fractions of the Microcystis population analyzed for microcystin concentration correlated with colony sim. In the small fraction (>30 <66 μm) microcystin was not detected. In the medium fraction (> 6 h < 100μm) lower microcystin concentrations were detected than in large fraction (>100μm) in which the highest microcystin concentrations were found.     

Prokaryotic Community Structure Driven by Salinity and Ionic Concentrations in Plateau Lakes of the Tibetan Plateau

The prokaryotic community composition and diversity and the distribution patterns at various taxonomic levels across gradients of salinity and physiochemical properties in the surface waters of seven plateau lakes in the Qaidam Basin, Tibetan Plateau, were evaluated using Illumina MiSeq sequencing. These lakes included Lakes Keluke (salinity, <1 g/liter), Qing (salinity, 5.5 to 6.6 g/liter), Tuosu (salinity, 24 to 35 g/liter), Dasugan (salinity, 30 to 33 g/liter), Gahai (salinity, 92 to 96 g/liter), Xiaochaidan (salinity, 94 to 99 g/liter), and Gasikule (salinity, 317 to 344 g/liter). The communities were dominated by Bacteria in lakes with salinities of <100 g/liter and by Archaea in Lake Gasikule. The clades At12OctB3 and Salinibacter, previously reported only in hypersaline environments, were found in a hyposaline lake (salinity, 5.5 to 6.6 g/liter) at an abundance of ∼1.0%, indicating their ecological plasticity. Salinity and the concentrations of the chemical ions whose concentrations covary with salinity (Mg²⁺, K⁺, Cl⁻, Na⁺, SO₄²⁻, and Ca²⁺) were found to be the primary environmental factors that directly or indirectly determined the composition and diversity at the level of individual clades as well as entire prokaryotic communities. The distribution patterns of two phyla, five classes, five orders, five families, and three genera were well predicted by salinity. The variation of the prokaryotic community structure also significantly correlated with the dissolved oxygen concentration, pH, the total nitrogen concentration, and the PO₄³⁻ concentration. Such correlations varied depending on the taxonomic level, demonstrating the importance of comprehensive correlation analyses at various taxonomic levels in evaluating the effects of environmental variable factors on prokaryotic community structures. Our findings clarify the distribution patterns of the prokaryotic community composition in plateau lakes at the levels of individual clades as well as whole communities along gradients of salinity and ionic concentrations.     

Quantitative Real-Time PCR for Determination of Microcystin Synthetase E Copy Numbers for Microcystis and Anabaena in Lakes

Cyanobacterial mass occurrences in freshwater lakes are generally formed by Anabaena, Microcystis, and Planktothrix, which may produce cyclic heptapeptide hepatotoxins, microcystins. Thus far, identification of the most potent microcystin producer in a lake has not been possible due to a lack of quantitative methods. The aim of this study was to identify the microcystin-producing genera and to determine the copy numbers of microcystin synthetase gene E (mcyE) in Lake Tuusulanjärvi and Lake Hiidenvesi in Finland by quantitative real-time PCR. The microcystin concentrations and cyanobacterial cell densities of these lakes were also determined. The microcystin concentrations correlated positively with the sum of Microcystis and Anabaena mcyE copy numbers from both Lake Tuusulanjärvi and Lake Hiidenvesi, indicating that mcyE gene copy numbers can be used as surrogates for hepatotoxic Microcystis and Anabaena. The main microcystin producer in Lake Tuusulanjärvi was Microcystis spp., since average Microcystis mcyE copy numbers were >30 times more abundant than those of Anabaena. Lake Hiidenvesi seemed to contain both nontoxic and toxic Anabaena as well as toxic Microcystis strains. Identifying the most potent microcystin producer in a lake could be valuable for designing lake restoration strategies, among other uses.     

Phylogenies of Microcystin-Producing Cyanobacteria in the Lower Laurentian Great Lakes Suggest Extensive Genetic Connectivity

Lake St. Clair is the smallest lake in the Laurentian Great Lakes system. MODIS satellite imagery suggests that high algal biomass events have occurred annually along the southern shore during late summer. In this study, we evaluated these events and tested the hypothesis that summer bloom material derived from Lake St. Clair may enter Lake Erie via the Detroit River and represent an overlooked source of potentially toxic Microcystis biomass to the western basin of Lake Erie. We conducted a seasonally and spatially resolved study carried out in the summer of 2013. Our goals were to: 1) track the development of the 2013 summer south-east shore bloom 2) conduct a spatial survey to characterize the extent of toxicity, taxonomic diversity of the total phytoplankton population and the phylogenetic diversity of potential MC-producing cyanobacteria (Microcystis, Planktothrix and Anabaena) during a high biomass event, and 3) compare the strains of potential MC-producers in Lake St. Clair with strains from Lake Erie and Lake Ontario. Our results demonstrated a clear predominance of cyanobacteria during a late August bloom event, primarily dominated by Microcystis, which we traced along the Lake St. Clair coastline downstream to the Detroit River''s outflow at Lake Erie. Microcystin levels exceeded the Province of Ontario Drinking Water Quality Standard (1.5 µg L⁻¹) for safe drinking water at most sites, reaching up to five times this level in some areas. Microcystis was the predominant microcystin producer, and all toxic Microcystis strains found in Lake St. Clair were genetically similar to toxic Microcystis strains found in lakes Erie and Ontario. These findings suggest extensive genetic connectivity among the three systems.     

Prebiotic Oligosaccharides: Comparative Evaluation Using In Vitro Cultures of Infants' Fecal Microbiomes

The objective of this study was to systematically assess the bifidogenic effect of three commonly used prebiotic products using in vitro cultures of infant fecal samples. Fresh stool samples collected from six term infants, each exclusively fed human milk (n = 3) or infant formula (n = 3), at 28 days of age were used as inocula. The following prebiotic products were added at concentrations applicable to infant formula: Vivinal GOS 15 (containing 28.5% galacto-oligosaccharide [GOS]) at 7.2 g/liter, Beneo HP (99.5% long-chain inulin [IN]) at 0.8 g/liter, Beneo Synergy 1 (enriched oligofructose and inulin [OF-IN]) at 4 g/liter, and a combination of Vivinal GOS 15 (7.2 g/liter) and Beneo HP (0.8 g/liter) (GOS-IN). The growth of total bacteria, Bifidobacterium, Bacteroides, Bifidobacterium longum, and Escherichia coli was quantified using specific quantitative PCR (qPCR). Bifidobacterium was also enumerated on selective Beerens agar plates, with representative colonies identified by sequencing of their 16S rRNA genes. Volatile fatty acids (VFA) and pH in the cultures were also determined. Irrespective of the feeding methods, the GOS product, either alone or in combination with Beneo HP, resulted in substantially higher growth of total bifidobacteria, and much of this growth was attributed to growth of B. longum. Beneo Synergy 1 also increased the abundance of total bifidobacteria and B. longum. Corresponding to the increases in these two bacterial groups, acetic acid concentrations were higher, while there was a trend of lower E. coli levels and pH. The lower pH and higher acetic acid concentration might be directly responsible for the lower E. coli population. At the concentrations studied, the GOS product was more bifidogenic and potent in inhibiting E. coli than the other products tested. These results suggest that supplementation of infant formula with GOS may increase intestinal bifidobacteria and benefit infant health.