Initial impacts of <Emphasis Type="Italic">Microcystis</Emphasis><Emphasis Type="Italic">aeruginosa</Emphasis> blooms on the aquatic food web in the San Francisco Estuary

The impact of the toxic cyanobacterium Microcystis aeruginosa on estuarine food web production in San Francisco Estuary is unknown. It is hypothesized that Microcystis contributed to a recent decline in pelagic organisms directly through its toxicity or indirectly through its impact on the food web after 1999. In order to evaluate this hypothesis, phytoplankton, cyanobacteria, zooplankton, and fish were collected biweekly at stations throughout the estuary in 2005. Concentrations of the tumor-promoting Microcystis toxin, microcystin, were measured in water, plankton, zooplankton, and fish by a protein phosphatase inhibition assay, and fish health was assessed by histopathology. Microcystis abundance was elevated in the surface layer of the western and central delta and reached a maximum of 32 × 10⁹ cells l⁻¹ at Old River in August. Its distribution across the estuary was correlated with a suite of phytoplankton and cyanobacteria species in the surface layer and 1 m depth including Aphanizomenon spp., Aulacoseira granulata, Bacillaria paradoxa, Rhodomonas spp., and Cryptomonas spp. Shifts in the phytoplankton community composition coincided with a decrease in the percentage of diatom and green algal carbon and increase in the percentage of cryptophyte carbon at 1 m depth. Maximum calanoid and cyclopoid copepod carbon coincided with elevated Microcystis abundance, but it was accompanied by a low cladocera to calanoid copepod ratio. Total microcystins were present at all levels of the food web and the greater total microcystins concentration in striped bass than their prey suggested toxins accumulated at higher trophic levels. Histopathology of fish liver tissue suggested the health of two common fish in the estuary, striped bass (Morone saxatilis), and Mississippi silversides (Menidia audens), was impacted by tumor-promoting substances, particularly at stations where total microcystins concentration was elevated. This study suggests that even at low abundance, Microcystis may impact estuarine fishery production through toxic and food web impacts at multiple trophic levels.     

Nitrogen-fixing cyanobacterium <Emphasis Type="Italic">Trichormus variabilis</Emphasis> of the Lake Baikal phytoplankton

A new filamentous cyanobacterial strain BAC 9610 was isolated from the lake Baikal pelagial. Data obtained by light, scanning, and transmission electron microscopy, along with 16S rRNA gene sequence analysis, allowed the bacterium identification as Trichormus variabilis, previously known as Anabaena variabilis. Trichormus is a cyanobacterial genus not presented in the list of Baikal plankton algae; A. variabilis also hasn’t been previously detected in Baikal phytoplankton. T. variabilis nitrogen fixation ability was demonstrated. The gene responsible for nitrogen fixation, nifH, was identified by PCR and was partially sequenced. No hepatotoxin synthesis genes were revealed in the strain.     

Genetic and morphological heterogeneity of Lake Baikal endemic gastropod <Emphasis Type="Italic">Benedictia fragilis</Emphasis> (Dybowski, 1875)

Baikal endemic Benedictia fragilis gastropods distributed in a wide range of depths (from sublittoral to abyssal) of three lake basins are studied. The analysis of the nucleotide sequence of the COI mitochondrial gene fragment and internal transcribed nuclear DNA spacer (ITS1) demonstrates that the studied gastropods are represented in Lake Baikal by three genetic groups. The results of the studies on genetic diversity, phenotypic traits, and distribution allow us to assume that the detected groups are incipient allopatric (geographical) species. On the basis of the data obtained and geological and climatic history of Baikal, possible pathways of the B. fragilis resettlement in the lake and the emergence of three genetic groups are hypothesized.     

Evidence for Recombination in the Microcystin Synthetase (<Emphasis Type="Italic">mcy</Emphasis>) Genes ofToxic Cyanobacteria <Emphasis Type="Italic">Microcystis</Emphasis><Emphasis Type="Bold">spp</Emphasis>

Recombination has been suggested to be an important factor for the genetic variation of bacterial genes, but few studies have dealt with intragenic recombination between the same or closely related species of cyanobacteria. Here we provide strong evidence for recombination in the microcystin synthetase (mcy) gene cluster of the toxic cyanobacteria Microcystis spp. This gene cluster contains 10 genes (mcyA to J) that encode a mixed polyketide synthase (PKS)/nonribosomal peptide synthetase (NRPS) complex. mcy gene sequences were determined for four selected regions (within mcyA, D, G, and J) within the mcy gene cluster from 1 Canadian and 10 Asian toxic Microcystis and compared with previously published mcy sequences. Split decomposition analysis indicated a reticulate phylogeny of mcyA, and several potential recombination tracts of mcyA were identified by the RDP analysis and a runs test implemented in GENECONV. In contrast, no recombination was detected in the mcyD, G, and J sequences. However, discrepancies among the four mcy gene genealogies were evident from the results of independent split decomposition analyses, which were further supported by incongruence length difference (ILD) tests. Taken together, these findings suggest that both intragenic and intergenic recombination within the mcy gene cluster contributes to the genetic diversity of the mcy genes of Microcystis spp.This article contains online supplementary material.     

The structure of <Emphasis Type="Italic">Dreissena polymorpha</Emphasis> (Pallas) biocenosis and the role of mollusk in roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis> (Linnaeus) feeding in Pleshcheevo Lake

As a result of studies on the macrozoobenthos composition of zebra mussel biocenosis in Lake Pleshcheevo carried out in 1996, a total of 44 species of bottom macroinvertebrates were found. The richest in terms of species composition were chironomids (17 species), oligochaetes (13 species), and mollusks and leeches (5 species each). After the introduction of zebra mussel Dreissena polymorpha (Pallas) into the lake, a natural biophilter was formed in the waterbody at depths of 4.5 to 9 m. This biophilter intercepts a substantial part of degrading macrophytes and allochtonous organic matter. Thus, the filter prevents the income of this organic matter in the lake’s profundal zone. Two roach Rutilus rutilus (Linnaeus) groups had inhabited Lake Pleshcheevo before dreissena introduction. Comparatively fast growing roach fed predominantly on mollusks of g. g. Valvata and Bithynia, while slowly growing roach fed mostly on zooplankton. After the introduction and mass development of D. polymorpha in the lake, roach started to consume these mollusks. This resulted in increases in the fish growth rate and maximum body size. Roach starts to feed on dreissena and other mollusks after reaching a body length of 15.0 cm. This relates to the third and the last change in the pharyngeal teeth shape, making possible for the fish to consume mollusks and to crush their shells.     

Dynamics of microcystin-degrading bacteria in mucilage of <Emphasis Type="Italic">Microcystis</Emphasis>

To reveal the process of degradation of hepatotoxic microcystin produced in Microcystis cells during the Microcystis bloom period, we used fluorescence in situ hybridization (FISH) to analyze the population dynamics of microcystin-degrading bacteria in Microcystis mucilage. We designed and applied an oligonucleotide probe targeted to the 16S rRNA sequence of strain Y2 of a microcystin-degrading bacterium (MCD-bacterium), which was isolated from Lake Suwa, Japan. In both the 1998 and 1999 tests, FISH clearly showed that MCD-bacteria existed in the mucilage and that, when a high concentration of cell-bound microcystin was detected, MCD-bacteria exceeded 10% of the sum of bacteria hybridized with group-specific probes. The concentration of MCD-bacteria was highest in summer 1998, when a toxic species, M. viridis, was dominant. There was a high correlation between the number of MCD-bacteria in the mucilage and the concentration of cell-bound microcystin in the lake. Our results suggest that MCD-bacteria responded to changes in the concentration of microcystin and degraded the microcystin when it was released from Microcystis cells. We also analyzed changes in the bacterial community structure associated with the Microcystis colonies by using domain- and group-specific oligonucleotide probes. Changes in the concentrations of the Cytophaga/Flavobacterium group and -Proteobacteria, which can degrade macromolecules derived from Microcystis cells, were synchronized with changes in the concentration of Microcystis. The results not only suggest the significant role of MCD-bacteria in detoxification, but also demonstrate a possible sequence of degradation from Microcystis cells to microcystin maintained in the cell, which is then carried out by bacterial consortia in the mucilage.     

Life history responses of <Emphasis Type="Italic">Daphnia longispina</Emphasis> to mosquitofish (<Emphasis Type="Italic">Gambusia holbrooki</Emphasis>) and pumpkinseed (<Emphasis Type="Italic">Lepomis gibbosus</Emphasis>) kairomones

In the present study, the effect of chemical cues from two fish species (mosquitofish and pumpkinseed), at different concentrations, was tested in life history experiments with Daphnia longispina. The two fish species used represent the most abundant planktivores of many Mediterranean shallow lakes (SW Europe), where the indigenous fish communities have been replaced by such exotic assemblages. Results have shown a similar response of D. longispina to both fish species: kairomones stimulated daphnids to produce more offspring, which resulted in higher fitness (r), relatively to a fishless control. Fish presence also induced an earlier first reproduction, a smaller size at maturity of daphnids, and the production of smaller-sized neonates. Significant correlations with fish concentration (indirect measure of fish kairomone concentration) were found for size at maturity and neonate size, for both fish species. These results are in accordance to the “positive response” observed by other authors, which represents a defence mechanism to face losses caused by fish predators. The chemically mediated size reduction of mature females and neonates is an adaptive response to the size-selective predation exerted by fish. Pumpkinseed introduction is very recent in the lake of origin of the daphnids used in the experiments and its kairomone produced similar effects to mosquitofish in the life history of D. longispina. These results are contrary to the existence of a species-specific kairomone and support the hypothesis of a general fish kairomone. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

Diversity of Freshwater <Emphasis Type="Italic">Thioploca</Emphasis> Species and Their Specific Association with Filamentous Bacteria of the Phylum <Emphasis Type="Italic">Chloroflexi</Emphasis>

Phylogenetic diversity among filamentous sulfur-oxidizing bacteria of the genus Thioploca inhabiting freshwater/brackish environments was analyzed in detail. The 16S rRNA gene sequence of Thioploca found in a freshwater lake in Japan, Lake Okotanpe, was identical to that of Thioploca from Lake Ogawara, a brackish lake. The samples of the two lakes could be differentiated by the sequences of their 23S rRNA genes and 16S–23S rRNA internal transcribed spacer (ITS) regions. The 23S rRNA-based phylogenetic relationships between Thioploca samples from four lakes (Lake Okotanpe, Lake Ogawara, Lake Biwa, and Lake Constance) were similar to those based on the 16S rRNA gene sequences. In addition, multiple types of the ITS sequences were obtained from Thioploca inhabiting Lake Okotanpe and Lake Constance. Variations within respective Thioploca populations were also observed in the analysis of the soxB gene, involved in sulfur oxidation. As major members of the sheath-associated microbial community, bacteria of the phylum Chloroflexi were consistently detected in the samples from different lakes. Fluorescence in situ hybridization revealed that they were filamentous and abundantly distributed within the sheaths of Thioploca.     

<Emphasis Type="Italic">Alona alsafadii</Emphasis> n. sp. from Yemen,a primitive,groundwater-dwelling member of the <Emphasis Type="Italic">A. karua</Emphasis>-group

The effect of algae on the production of musty-smelling compounds by actinomycetes was studied. Streptomyces spp., causing intensive musty odor, were isolated from hypertrophic Lake Kasumigaura and cultured in association with algae from the same lake. Isolate E and I effectively utilized the cyanobacteria, Microcystis aeruginosa and Anabaena spiroides, and the diatom, Synedra acus, as a carbon source and produced a musty-smelling 2-methylisoborneol in the shaken sediment cultures. High populations of algae and actinomycetes, and aerobic condition in the sediment seem responsible for the occurrence of musty odor in Lake Kasumigaura.     

Characteristics of melanomacrophage centers in the liver and spleen of the roach Rutilus rutilus (Cypriniformes: Cyprinidae) in Lake Kotokel during the Haff disease outbreak

Characteristics of morphology and number of melanomacrophage centers (MMCs) in the liver and spleen of the roach Rutilus rutilus and the amount of pigments in MMCs during the Haff disease outbreak and the death of fish in Lake Kotokel in relation to these parameters in the roach from Lake Baikal are described. Pathological changes in the microvasculature and parenchyma in the liver of the roach from Lake Kotokel were found. The area of melanomacrophage centers in the liver of the roach from this lake was significantly smaller, whereas the number and size of these centers in the spleen was significantly larger than in the roaches from Lake Baikal. Among the pigments studied, the strongest response to the content of this toxin in the water body was shown by hemosiderin. An increase in its amount in the spleen MMCs testifies to an enhanced degradation of erythrocytes and iron release, which may be caused by the damage of cells of the erythrocyte lineage by the toxin.     

Accumulation and depuration of microcystin produced by the cyanobacterium <Emphasis Type="Italic">Microcystis</Emphasis> in a freshwater snail

Seasonal changes in microcystin concentrations in a resident snail (Sinotaia histrica) and an edible clam (Corbicula sandai) in Lake Biwa were surveyed. To clarify both the accumulation and depuration of microcystins, experimental studies with microcystin were also carried out on the snail. In the field investigation, microcystin was detected from the hepatopancreas and intestine of S. histrica (up to 3.2µgg^–1 dry weight and 19.5µgg^–1 dry weight, respectively); however, no microcystin was detected in the hepatopancreas of C. sandai. In the laboratory experiment, the microcystin-LR concentration in the hepatopancreas of S. histrica reached a value of 436µgg^–1 dry weight on day 10 of 15 days of uptake, and a high value persisted despite a depuration period of 15 days. The depuration rate constant of microcystin and its biological half-life were 0.0828 day^–1 and 8.4 days, respectively. These results indicate that S. histrica has a high ability to accumulate microcystin in its tissue. Because S. histrica is predated by fish and water fowl, it is likely to play an important role as a vector for microcystin in lakes with dense blooms of toxic cyanobacteria.     

Food composition and feeding rate of the lake Baikal invader <Emphasis Type="Italic">Gmelinoides fasciatus</Emphasis> (Stebbing, 1899) in Lake Ladoga

The feeding of the Baikal invader Gmelinoiudes fasciatus (Stebbing, 1899) in Lake Ladoga has been studied based on field and experimental data. The food spectrum of this amphipod was revealed; it includes many plant and animal species. Differences in the feeding of G. fasciatus living in different littoral biotopes were shown. The ecological and physiological rations of this species were determined. The effect that the bottom substrate has on the feeding rate of the amphipod was estimated. The introduction of G. fasciatus into Lake Ladoga provided a more adequate utilization of the energy accumulated by the primary producers and its transfer to higher trophic levels. This Baikal amphipod occupied a niche that had been, in effect, vacant. This made the invader capable of maintaining the quantitative parameters of its population at a very high level, and it largely reorganized the flow of matter and energy in the littoral zone of the lake.     

<Emphasis Type="Italic">Coregonus peled</Emphasis> (Gmelin) transplanted into Ulaagchny Khar lake (western mongolia) showed no evidence of hybridization with other introduced <Emphasis Type="Italic">Coregonus</Emphasis> species

Coregonus peled (Gmelin) (Teleostei: Salmoniformes: Coregonidae), which is considered an important object of coldwater aquaculture, had been successfully introduced into an enclosed Western Mongolian lake Ulaagchny Khar in the early 1980s. At the same time larvae of two other Coregonus species—Baikal omul C. migratorius (Georgi) and least cisco C. sardinella Valenciennes—had also been released into the lake. Baikal omul was then reported as a naturalized species. This might have caused interspecific hybridization and gene introgression. Identification of coregonids by morphology can be problematic, so to determine which species was dominant in the lake (we presumed it was peled) and if its gene pool was affected by other introduced Coregonus species we sampled 40 individuals and analyzed them by sequencing a fragment of mtDNA cyt b and by allozyme electrophoresis. The analysis showed that all the fish belonged to C. peled with no evidence of admixture from other coregonid species. Taking into account mass release of both species in 1980s, it is evident that naturalization of peled in the lake was much more successful than that of Baikal omul.     

Composition of leucocytes of the head kidney of the crucian carp (<Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>, Cypriniformes: Cyprinidae) as affected by invasion of cestode <Emphasis Type="Italic">Digramma interrupta</Emphasis> (Cestoda; Pseudophyllidea)

The composition of leucocytes of the head kidney is studied in the crucian carps (Carassius auratus) either contaminated or uncontaminated with Digramma interrupta. The composition of leucocytes in the pronephros of the crucian carp from Lake Baikal basin has a lymphoid character. Compared to the crucian carp from the European part of Russia, in the fish from Baikal the granulocytopoetic processes are more pronounced. This is proved by the high content of young forms of granulocytes. In the fish infected with digramma, the immune suppression of proliferation of blasts and young forms of eosinophils was revealed. On the other hand, the inflammatory and humoral specific immune reactions are enhanced. Partial suppression of the immune response of C. auratus to invasion by D. interrupta facilitates development of the parasite.     

Evidence of repeated long-distance movements by lake charr <Emphasis Type="Italic">Salvelinus namaycush</Emphasis> in Lake Huron

Movements and dispersal distances of acoustically-tagged adult lake charr Salvelinus namaycush were estimated based on detections at acoustic receivers in Lake Huron during 2010–2014. Most lake charr were detected only at receivers proximate to their release location or were not detected at all, but 3–9% of tagged lake charr were detected at receivers located over 100 km from their release location. Several fish made extensive repeated migrations within the lake, some at the scale of the entire main basin. Our observations show that some lake charr individuals repeat a similar pattern each year of moving long distances, and some fish were observed to show annual fidelity to presumed foraging sites in the spring at a spatial scale of approximately 200 km. Our telemetry-based estimates were minimum estimates of dispersal, as the placement of receivers within Lake Huron was not optimal for detection of lake charr and did not cover the majority of the lake. Further study of long-distance movement in lake charr is necessary to fully understand the implications of this behavior to lake charr ecology, population dynamics, and management in the Great Lakes.     

Colorless sulfur bacteria <Emphasis Type="Italic">Thioploca</Emphasis> from different sites in Lake Baikal

The colorless sulfur bacteria Thioploca spp. found in Lake Baikal are probably a marker for the influx of subterranean mineralized fluids. Bacteria act as a biological filter; by consuming sulfide in their metabolism, they detoxicate it and maintain the purity of Lake Baikal’s water. The bacteria were investigated by various techniques. According to analysis of the 16S rRNA gene fragment, Thioploca sp. from Frolikha Bay, Baikal belongs to the clade of freshwater species found in Lake Biwa and Lake Constance; it is most closely related to Thioploca ingrica.     

Studies on mineral phosphate solubilization by cyanobacteria <Emphasis Type="Italic">Westiellopsis</Emphasis> and <Emphasis Type="Italic">Anabaena</Emphasis>

Two diazotrophic cyanobacteria, Westiellopsis prolifica and Anabaena variabilis were evaluated for elucidating the possible mechanism of mineral phosphate solubilization. Phosphate starved cyanobacteria evaluated for the presence of organic acids, extracellular compounds or enzymes that might have been produced and promoted the mineral phosphate solubilization with Mussorie Rock Phosphate and Tricalcium Phosphate as substrates. Both the cultures did not reveal production of organic acids throughout the incubation period when checked for decrease in media pH of the media and thin layer chromatography. Thin layer chromatography of culture filtrates showed the presence of hydrocarbon like compound. Further analysis of the culture filtrates with gas liquid chromatography, a single peak near to the retention time of 7.6 was observed in all extracts of culture filtrates irrespective of phosphate source. UV-visible spectra of culture filtrates revealed the absorption maxima of 276 nm. Gas chromatographic-mass spectrometric analysis of culture filtrates showed most intense peak in the electron impact (EI) ionization was at m/z 149 and molecular ion peaks at m/z 207 and 167, inferring the presence of phthalic acid. Among the mechanisms in mineral phosphate solubilization, it was evident that these cyanobacteria used phthalic acid as possible mode of P solubilization.     

Development of subfossil <Emphasis Type="Italic">Daphnia</Emphasis> and <Emphasis Type="Italic">Chaoborus</Emphasis> assemblages in relation to progressive acidification and fish community alterations in SW Sweden

Dated sediment cores from acidified and fishless Lake Gaffeln and Lake Härsvatten, SW Sweden, were analyzed for Daphnia ephippia and Chaoborus mandibles to test whether acidification history and fish extirpations could be reconstructed in a paleo-study using these easily identifiable animal remains. According to monitoring data fish were lost in both lakes from the 1950s to the 1970s. Progressive acidification prior to monitoring was confirmed by a gradual decrease and eventual loss of Daphnia ephippia in both study lakes during the first half of the twentieth century. In Lake Gaffeln mandibles of C. obscuripes appeared immediately after fish loss in 1973, and the regular presence of this species confirmed the succeeding fishless state of this lake. In Lake Härsvatten sediments C. obscuripes appeared only recently, i.e. three decades after fish extirpation, showing that the absence of C. obscuripes mandibles is not a trustworthy indicator of fish presence. Hence, the appearance of C. obscuripes was not temporally related to fish loss but confirmed the present fishless condition. Known historical presence of cyprinid fish in Lake Gaffeln was confirmed by a significantly higher proportion of fragmented mandibles of C. flavicans compared to the historically cyprinid-free Lake Härsvatten. In addition, both lake profiles displayed zero-proportions of fragmented mandibles during fishless periods. We conclude that acidification history and fish extirpations can be inferred by integrated studies on subfossil Daphnia ephippia and Chaoborus mandibles. However, during extreme ultra-oligotrophic conditions in acidified clear-water lakes subfossil Chaoborus mandibles may be too scarce to infer fish absence/presence.     

Characterization of paralytic shellfish toxins in seawater and sardines (<Emphasis Type="Italic">Sardina pilchardus</Emphasis>) during blooms of <Emphasis Type="Italic">Gymnodinium catenatum</Emphasis>

The re-emergence of Gymnodinum catenatum blooms after a 10 year hiatus of absence initiated the present investigation. This study aims to evaluate the exposure of small pelagic fishes to paralytic shellfish toxins (PST) during blooms of G. catenatum. Sardines (Sardina pilchardus) were selected as a representative fish species. In order to assess toxin availability to fish, both intracellular PSTs (toxin retained within the algal cells) and extracellular PSTs (toxin found in seawater outside algal cells) were quantified, as well as toxin levels within three fish tissue matrices (viscera, muscle and brain). During the study period, the highest cell densities of G. catenatum reached 2.5 × 10⁴ cells l⁻¹ and intracellular PST levels ranged from 3.4 to 398 ng STXeq l⁻¹ as detected via an enzyme linked immunosorbent assay (ELISA). Measurable extracellular PSTs were also detected in seawater (0.2–1.1 μg STXeq l⁻¹) for the first time in Atlantic waters. The PST profile in G. catenatum was determined via high performance liquid chromatography with fluorescence detection (HPLC-FLD) and consisted mostly of sulfocarbamoyl (C1+2, B1) and decarbamoyl (dcSTX, dcGTX2+3, dcNEO) toxins. The observed profile was similar to that reported previously in G. catenatum blooms in this region before the 10-year hiatus. Sardines, planktivorous fish that ingest a large number of phytoplankton cells, were found to contain PSTs in the viscera, reaching a maximum of 531 μg STXeq kg⁻¹. PSTs were not detected in corresponding muscle or brain tissues. The PST profile characterized in sardine samples consisted of the same sulfocarbamoyl and decarbamoyl toxins found in the algal prey with minor differences in relative abundance of each toxin. Overall, the data suggest that significant biotransformation of PSTs does not occur in sardines. Therefore, planktivorous fish may be a good tracer for the occurrence of offshore G. catenatum blooms and the associated PSTs produced by these algae.