PCR-DGGE Fingerprinting Analysis of Plankton Communities and Its Relationship to Lake Trophic Status

Plankton communities in eight lakes of different trophic status near Yangtze, China were characterized by using denatured gradient gel electrophoresis (DGGE). Various water quality parameters were also measured at each collection site. Following extraction of DNA from plankton communities, 16S rRNA and 18S rRNA genes were amplified with specific primers for prokaryotes and eukaryotes, respectively; DNA profiles were developed by DGGE. The plankton community of each lake had its own distinct DNA profile. The total number of bands identified at 34 sampling stations ranged from 37 to 111. Both prokaryotes and eukaryotes displayed complex fingerprints composed of a large number of bands: 16 to 59 bands were obtained with the prokaryotic primer set; 21 to 52 bands for the eukaryotic primer set. The DGGE-patterns were analyzed in relation to water quality parameters by canonical correspondence analysis (CCA). Temperature, pH, alkalinity, and the concentration of COD, TP and TN were strongly correlated with the DGGE patterns. The parameters that demonstrated a strong correlation to the DGGE fingerprints of the plankton community differed among lakes, suggesting that differences in the DGGE fingerprints were due mainly to lake trophic status. Results of the present study suggest that PCR-DGGE fingerprinting is an effective and precise method of identifying changes to plankton community composition, and therefore could be a useful ecological tool for monitoring the response of aquatic ecosystems to environmental perturbations.

Mitigation of lake eutrophication: Loosen nitrogen control and focus on phosphorus abatement

w Traditionally, nitrogen control is generally considered an important component of reducing lake eutrophication and cyanobacteria blooms. However, this viewpoint is refuted recently by researchers in China and North America. In the present paper, the traditional viewpoint of nitrogen control is pointed out to lack a scientific basis: the N/P hypothesis is just a subjective assumption; bottle bioassay experiments fail to simulate the natural process of nitrogen fixation. Our multi-year comparative research in more than 40 Yangtze lakes indicates that phosphorus is the key factor determining phytoplankton growth regardless of nitrogen concentrations and that total phytoplankton biomass is determined by total phosphorus and not by total nitrogen concentrations. These results imply that, in the field, nitrogen control will not decrease phytoplankton biomass. This finding is supported by a long-term whole-lake experiment from North America. These outcomes can be generalized in terms that a reduction in nitrogen loading may not decrease the biomass of total phytoplankton as it can stimulate blooms of nitrogen-fixing cyanobacteria. To mitigate eutrophication, it is not nitrogen but phosphorus that should be reduced, unless nitrogen concentrations are too high to induce direct toxic impacts on human beings or other organisms. Finally, details are provided on how to reduce controls on nitrogen and how to mitigate eutrophication. (C) 2009 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

Seasonal Variations in Chlorophyll a Concentrations in Relation to Potentials of Sediment Phosphate Release by Different Mechanisms in a Large Chinese Shallow Eutrophic Lake (Lake Taihu)

The relationship between chlorophyll a and fractionation of sediment phosphorus, inorganic phosphate-solubilizing bacteria (IPB), and organic phosphate-mineralizing bacteria (OPB) was evaluated in a large Chinese shallow eutrophic lake (Lake Taihu) and its embayment (Wuli Bay). At the three study sites, the increase of chlorophyll a concentrations in April paralleled those of the iron bound phosphate accounting for major portion of sediment inorganic phosphate, and in June significantly higher OPB and IPB numbers (especially OPB) in sediment were main contributors to the peaks of chlorophyll a concentration. Even though IPB peaked from February to June, it should serve as an unimportant P source due to the irrelevancy with chlorophyll a and soluble reactive phosphorus (SRP). By contrast, at the other site in the embayment, the calcium-bound phosphate was predominant and solid, which was difficult to be released, and neither IPB nor OPB were detectable in the sediment, indicating weak potential for phosphorus release from the sediment, which was reflected in the small seasonal variation in SRP concentration in water column. Hence, the extents to which the three general mechanisms behind phosphate release from sediment (desorption of iron bound phosphate, solubilization by IPB and enzymatic hydrolysis by OPB) operated were different depending on seasons and sites in Lake Taihu, they may jointly drive phosphate release and accelerate the eutrophication processes.

Determination of microcystin-LR and its metabolites in snail (Bellamya aeruginosa), shrimp (Macrobrachium nipponensis) and silver carp (Hypophthalmichthys molitrix) from Lake Taihu, China

This paper describes seasonal changes of microcystin-LR (MC-LR) and its glutathione (MC-LR-GSH) and cysteine conjugates (MC-LR-Cys) in three aquatic animals - snail (Bellamya aeruginosa), shrimp (fMacrobrachium nipponensis) and silver carp (Hypophthalmichthys molitrix) collected from Lake Taihu, China. MC-LR, MC-LR-GSH, and MC-LR-Cys were determined by liquid chromatography electrospray ionization mass spectrum (LC-ESI-MS). The mean MC-LR concentrations in the hepatopancreas of snail and shrimp and liver of silver carp were 6.61, 0.24, and 0.027 mu g g(-1) dry weight (DW), respectively: while the average MC-LR-Cys concentrations were 0.50, 0.97, and 5.72 mu g g(-1) DW, respectively. MC-LR-GSH was usually not detectable in these samples. The above results suggest that: (1) in aquatic animals, especially fish, the main excretion form of MC-LR could be MC-LR-Cys, but not MC-LR-GSH, whereas MC-LR-Cys might play an important role in detoxication of MC-LR and (2) that efficiency of MC-LR-Cys formation differs among species. The main detoxication pathway of MC-LR in aquatic animals is suggested as follows: when MC-LR enters into liver/hepatopancreas, it firstly conjugates with polypeptide or protein (including GSH, PP-1 and 2A) containing Cys residues, perhaps also some free cysteine; subsequently, MC-LR-Cys is degraded from these polypeptide or protein; and finally is excreted from animals by the compound of MC-LR-Cys. (C) 2009 Elsevier Ltd. All rights reserved.

The youngest split in sympatric schizothoracine fish (Cyprinidae) is shaped by ecological adaptations in a Tibetan Plateau glacier lake

Although new empirical evidence shows that sympatric speciation has occurred in some species, there are few indisputable model organisms for this process of speciation. The two subspecies (Gymnocypris eckloni eckloni and G. e. scoliostomus) of the schizothoracine Gymnocypris fish species complex from a small glacier lake in the Tibetan Plateau, Lake Sunmcuo, fit several of the key characteristics of the sympatric speciation model. We used combined mitochondrial control region sequences and the cytochrome b gene (1894 bp) to address the phylogenetics and population genetics of 232 specimens of G. e. eckloni and G. e. scoliostomus, as well as all of its closely related sister species. We found that: (i) a total of four old lineages were uncovered in the widespread G. e. eckloni, of which only one was shown to be shared with all G. e. scoliostomus individuals and (ii) the new subspecies (G. e. scoliostomus) evolved in Lake Sunmcuo from the ancestral G. e. eckloni population within approximately 0.057 Ma. These two taxa of the species complex are morphologically distinct, and reproductive isolation is further suggested. Ecological disruptive selection based on morphological traits (e.g. mouth cleft characters) and food utilization may be a mechanism of incipient speciation of two sympatric populations within Lake Sunmcuo. This study provides the first genetic evidence for sympatric speciation in the schizothoracine fish.

Phosphorus in suspended matter and sediments of a hypertrophic lake. A case study: Lake Dianchi, China

From June 2004 to December 2004, Lake Dianchi, which had large scale of cyanobacterial blooms was investigated in order to study P-fractionation in the suspended matter and the sediment. The investigation improves our understanding of phosphorus in Lake Dianchi and the relationship between phosphorus and cyanobacterial blooms. It contributes to the available literature on the behavior of P in hypertrophic lakes. The distribution of P-fractions in Lake Dianchi was not uniform from northwest to south, but was closely related to the trophic status of the whole lake. The concentrations of total phosphorus, labile P (NH4Cl-P), Organic P (NaOH-NRP) and loss on ignition in suspended matter were positively correlated with the strength of cyanobacterial blooms. Total phosphorus in suspended matter was relatively stable for almost half an year and closely related to Chl. a concentration. The main content of organic phosphorus is in the cyanobacterial blooms. The concentrations of phosphorus bound to metal oxides and carbonates (NaOH-SRP and HCl-P) in sediment were similar to NaOH-SRP and HCl-P in the corresponding suspended matter. The latter two forms of P in suspended matter were not affected by cyanobacterial blooms, indicating that the inorganic phosphorus is derived from the sediment after resuspension from the sediment due to wind and wave action. The contribution of the different P-fractions to TP in sediment and in suspended matter indicates that NH4Cl-P in the suspended matter is an important buffer for maintaining dissolved phosphorus in water.

Diversity and abundance of ammonia-oxidizing bacteria in eutrophic and oligotrophic basins of a shallow Chinese lake (Lake Donghu)

Classical cultivation and molecular methods based on the ammonia monooxygenase gene (amoA) were used to study the abundance and diversity of beta-proteobacterial ammonia-oxidizing bacteria (AOB) in lake sediments. The eutrophic and oligotrophic basins of a Chinese shallow lake (Lake Donghu), in terms of ammonium (NH4+) concentrations, were sampled. The AOB number was significantly lower in the oligotrophic basin, but significantly higher in the eutrophic basin. In addition, using restriction fragment length polymorphism targeting the amoA, ten restriction patterns including six unique ones were found in the eutrophic basin, while five patterns were observed in the oligotrophic basin with only one unique restriction group. Phylogenetic analysis for AOB revealed that Nitrosomonas oligotropha- and Nitrosomonas ureae-related AOB and Nitrosospira-affiliated AOB were ubiquitous; the former dominated in the eutrophic basin (87.2%), while the latter dominated in the oligotrophic basin (65.5%). Furthermore, Nitrosomonas communis-related AOB was only detected in the eutrophic basin, at a small proportion (3.2%). These results indicate significant selection and adaptation of sediment AOB in lakes with differing trophic status. (C) 2009 Elsevier Masson SAS. All rights reserved.

VERTICAL DISTRIBUTIONS OF SIZE-FRACTIONATED IRON IN THREE TYPICAL BAYS OF LAKE DIANCHI (CHINA)

In order to elucidate the vertical distributions of iron in three typical bays (Haigeng bay, Macun bay and Haidong bay) of Lake Dianchi (China), the investigation was conducted on March, 2003. Results showed that the vertical distributions were influenced by monsoon, cyanobacterial bloom and water depth as well as sediment resuspension, which indicated that their translocations and transformations were decided by geographical and physical as well as chemical and biological characteristics.

Assessment effects of cage culture on nitrogen and phosphorus dynamics in relation to fallowing in a shallow lake in China

Nitrogen and phosphorus dynamics in relation to fallowing in a fish cage farm was investigated in a shallow lake in China. Four sampling sites were set: beneath the cages, at the cage sides, and 50 and 100 m east of the cage farm. Total nitrogen (TN) and total phosphorus (TP) in lake water and sediment were analyzed during a 2-year rearing cycle. The cage culture had a fish yield of 16.3-39.2 tonnes in the study period. Based on the mass balance equation, 1533-3084 kg TN and 339-697 kg TP were contributed to the lake environment. Nitrogen and phosphorous concentrations showed greater increase in the first culture period than in the second rearing cycle. No obvious changes were found at the sampling sites 50 and 100 m east of the cages during the study periods. Main impacts were found close to the cages (beneath the cages and at the cage side); the sampling points at the cage side showed relatively high TN and TP sedimentation. After 3 months of fallowing, water TN and TP decreased significantly but the sediment TN and TP contents remained high. Therefore, recovery seems to happen during fallowing but attention should be paid to whether the culture continues to operate in the future.

Simultaneous determination of microcystin contaminations in various vertebrates (fish, turtle, duck and water bird) from a large eutrophic Chinese lake, Lake Taihu, with toxic Microcystis blooms

This is the first to conduct Simultaneous determination of microcystin (MC) contaminations in multi-groups of vertebrates (fish, turtle, duck and water bird) from Lake Taihu with Microcystis blooms. MCs (-RR, -YR, -LR) in Microcystis scum was 328 mu g g(-1) DW. MCs reached 235 mu g g(-1) DW in intestinal contents of phytoplanktivorous silver carp, but never exceeded 0.1 mu g g(-1) DW in intestinal contents of other animals. The highest MC content in liver of fish was in Carassius auratus (150 ng g(-1) DW), followed by silver carp and Culter ilishaeformis, whereas the lowest was in common carp (3 ng g(-1) DW). In livers of turtle, duck and water bird, MC content ranged from 18 to 30 ng g(-1) DW. High MC level was found in the gonad, egg yolk and egg white of Nycticorax nycticorax and Anas platyrhynchos, suggesting the potential effect of MCs on water bird and duck embryos. High MC contents were identified for the first time in the spleens of N. nycticorax and A. platyrhynchos (6.850 and 9.462 ng g(-1) DW, respectively), indicating a different organotropism of MCs in birds. Lakes with deaths of turtles or water birds in the literatures had a considerably higher MC content in both cyanobacteria and wildlife than Lake Taihu, indicating that toxicity of cyanobacteria may determine accumulation level of MCs and consequently fates of aquatic wildlife. (C) 2009 Elsevier B.V. All rights reserved.

Isolation of a novel cyanophage infectious to the filamentous cyanobacterium Planktothrix agardhii (Cyanophyceae) from Lake Donghu, China

A previously unknown cyanophage, PaV-LD (Planktothrix agardhii Virus isolated from Lake Donghu), which causes lysis of the bloom-forming filamentous cyanobacterium P. agardhii, was isolated from Lake Donghu, Wuhan, China. PaV-LD only lysed P. agardhii strains isolated from Lake Donghu and not those isolated from other lakes. The PaV-LD particle has an icosahedral, non-tailed structure, ca. 70 to 85 nm (mean +/- SD = 76 +/- 6 nm) in diameter. PaV-LD was stable at freezing temperature, but lost its infectivity at temperatures >50 degrees C. Lysis of host cells was delayed about 3 d after the PaV-LD treatment with chloroform, and the virus was inactivated by exposure to low pH (<= 4). The latent period and burst size of the PaV-LD were estimated to be 48 to 72 h and about 340 infectious units per cell, respectively. The regrowth cultures of surviving host filaments were not lysed by the PaV-LD suspension. To our knowledge, this is the first isolation and cultivation of a virus infectious to the filamentous bloom-forming cyanobacterium Planktothrix from a freshwater lake.

Transfer, distribution and bioaccumulation of microcystins in the aquatic food web in Lake Taihu, China, with potential risks to human health

In this paper, accumulation and distribution of microcystins (MCs) was examined monthly in six species of fish with different trophic levels in Meiliang Bay, Lake Taihu, China, from June to November 2005, Microcystins were analyzed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). Average recoveries of spiked fish samples were 67.7% for MC-RR, 85.3% for MC-YR, and 88.6% for MC-LR. The MCs (MC-RR+MC-YR+MC-LR) concentration in liver and gut content was highest in phytoplanktivorous fish, followed by omnivorous fish, and was lowest in carnivorous fish; while MCs concentration in muscle was highest in omnivorous fish, followed by phytoplanktivorous fish, and was lowest in carnivorous fish. This is the first study reporting MCs accumulation in the gonad of fish in field. The main uptake of MC-YR in fish seems to be through the gills from the dissolved MCs. The WHO limit for tolerable daily intake was exceeded only in common carp muscle. (C) 2008 Elsevier B.V. All rights reserved.

EXPERIMENTS ON AQUATIC ECOSYSTEM REGIME SHIFT IN ENCLOSURES NEAR LAKE DIANCHI, CHINA

To discover how a lake converts from a turbid state to clean state, and what drives this process, we constructed controlled enclosure ecosystems and used the ecological remediation method to force ecosystems to convert from the turbid state to the clean state. Our results show that the driving forces include temperature., macrophyte, silver carp and mussel, which form a combined force to drive the controlled ecosystem to switch. There is a threshold existing in treated enclosure ecosystem during the conversion from turbid to clean state. When TP <0.09 mg.L-1, Chl-a <0.036 mg.L-1, transparency >62 cm, TN <2.15 mg.L-1, CODMn <13.7 mg.L-1, tubidity <10, and the number of algal cells <10(6) cells.L-1, the treated ecosystem changes sharply from turbid to clean state. The conversion process can be divided into three phases: turbid state, clean-turbid transitional state as well as clean state, and described with the power function Y = a*X-b (where Y is water parameter, X is time, a and b are constants), which indicates that the shift in the enclosure ecosystem from turbid to clean state is discontinuous.

Genetic Diversity of the Plankton Community and Its Relation to Taxonomic Composition and Environmental Factors in Lake Xiliang

Genetic diversity of the plankton community in Lake Xiliang was depicted by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting. Seventy-seven bands (33 of 16S rDNA and 44 of 18S rDNA) were detected, sixty-two planktonic taxa were identified in six sample stations in November 2007. The most common taxa were Ceratium hirundinella, Bdelloidea, Keratella cochlearis, Polyarthra trigla, and copepod nauplii. Based on environmental factors, taxonomic composition, and PCR-DGGE fingerprinting, unweighted pair-group method using arithmetic averages clustering and principal components analysis were used to analyze habitat similarities. There was distinct spatial heterogeneity in Lake Xiliang, and the genetic diversity of the plankton community was closely related to taxonomic composition and environmental factors.

Food Consumption by In Situ Pen-Cultured Planktivorous Fishes and Effects on an Algal Bloom in Lake Taihu, China

Silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) were used as a new pen-cultureed biomanipulation technique to control algal blooms in Meiliang Bay of Lake Taihu. In order to evaluate the capacity of these two fishes to decrease algal blooms, diel feeding samplings were carried out in May (without algal blooms) and September (with algal blooms) in 2005. Based on estimated food consumption by the Elliott-Persson model, silver carp increased daily food consumption from 2.07 g dry weight per 100 g wet body weight in May before the outbreak of algal blooms to 4.98 g dry weight per 100 g wet body weight in September during algal blooms outbreak. However, no obvious variation of food consumption was observed in bighead carp during the study period. This species 1.88 and 1.54 g dry weight of plankton per 100 g wet body weight in May and September, respectively. Silver carp had a higher feeding capacity for plankton than bighead carp. Biotic factors (i.e., fish size and conspecific competition with natural species in the lake) may affect the feeding behaviors of both carps as well as seasonal variation of plankton communities in the pen.