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.

Plankton Community Succession in Artificial Systems Subjected to Cyanobacterial Blooms Removal using Chitosan-Modified Soils

Using artificial systems to simulate natural lake environments with cyanobacterial blooms, we investigated plankton community succession by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting and morphological method. With this approach, we explored potential ecological effects of a newly developed cyanobacterial blooms removal method using chitosan-modified soils. Results of PCR-DGGE and morphological identification showed that plankton communities in the four test systems were nearly identical at the beginning of the experiment. After applying the newly developed and standard removal methods, there was a shift in community composition, but neither chemical conditions nor plankton succession were significantly affected by the cyanobacteria removal process. The planted Vallisneria natans successfully recovered after cyanobacteria removal, whereas that in the box without removal process did not. Additionally, canonical correspondence analysis indicated that other than for zooplankton abundance, total phosphorus was the most important environmental predictor of planktonic composition. The present study and others suggest that dealing with cyanobacteria removal using chitosan-modified soils can play an important role in controlling cyanobacterial blooms in eutrophicated freshwater systems.

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.

Genetic diversity of plankton community as depicted by PCR-DGGE fingerprinting and its relation to morphological composition and environmental factors in lake donghu

To collect information about the genetic diversity of the plankton community and to study how plankton respond to environmental conditions, plankton samples were collected from five stations representing different trophic levels in a shallow, eutrophic lake (Lake Donghu), and investigated by PCR-DGGE fingerprinting. A total of 100 bands (61 of 16S rDNA bands and 39 of 18S rDNA bands) were detected. The DGGE bands unique to any single station accounted for 38% of the total bands, whereas common bands detected at all five stations accounted for only 11%. Using UPGMA clustering and MDS ordination of DGGE fingerprints, stations I and II were found to initially group together into one cluster, which was later joined by station V. Stations III and IV were isolated into two separate groups of one station each. Some differences in grouping relationships were found when analysis was completed on the basis of chemical characteristics and morphological composition, with zooplankton composition showing the greatest variability. However, the most similar stations (I and II) were always initially grouped into one cluster. Moreover, stations that exhibited the same or similar trophic level (stations III and IV), but different concentrations of heavy metals, were further differentiated by the DGGE method. Results of the present study indicated that PCR-DGGE fingerprinting was more sensitive than the traditional methods, as other studies suggested. Additionally, PCR-DGGE appears to be more appropriate for diversity characterization of the plankton community, as it is more canonical, systematic, and effective. Most importantly, fingerprinting results are more convenient for the comparative analyses between different studies. Therefore, the use of the described fingerprinting analysis may provide an operable and sensitive biomonitoring approach to identify critical, and potentially negative, stress within an aquatic ecosystem.

DNA polymorphism of the plankton community and its relationships to species composition in the Three Gorges Reservoir Region of the Yangtze River

We examined DNA polymorphism of the plankton community in the Three Gorges Reservoir Region of Yangtze River and studied its relationships to species composition. Samples of the plankton community were collected from nine sampling sites and analyzed by random amplified polymorphic DNA (RAPD). Nine of 60 screened primers generated a total of 88 observable 180 to 1400 by bands, all of which were polymorphic. Cluster analysis of the resulting binary format from DNA banding patterns grouped the target communities into three clusters. The topology of the constructed diagram from species composition data was generally similar to that based on RAPD markers.

Effects of the phytoplanktivorous silver carp (Hypophthalmichthys molitrixon) on plankton and the hepatotoxic microcystins in an enclosure experiment in a eutrophic lake, Lake Shichahal in Beijing

The responses of nutrients, water transparency, zooplankton, phytoplankton and microcystins to a gradient of silver carp biomass (0, 18, 55, 110 g/m(3)) were assessed using enclosures in Lake Shichahai (Beijing). Picophytoplankton biomass increased with increasing fish stocking density (r=0.64, p=0.09). Silver carp significantly depressed zooplankton biomass, and thus, zooplankton grazing was too low to control phytoplankton. Intracellular microcystin (MC) content in the enclosures was correlated only to Microcystis biomass in the present study. Microcystis spp. biomass and intracellular microcystins content were much higher in lake water than those of enclosures with and without stocking fish. Stocking of silver carp could be an appropriate in highly productive Lake Shichahai, which naturally lacks of large cladoceran zooplankton. A fish stocking density of 55 g/m(3) was most efficient at controlling Microcystis blooms and increasing water clarity. Mean extracellular MC concentration in the lake water was almost the same with that of the enclosures with fish. (c) 2006 Elsevier B.V. All rights reserved.

Difflugia mulanensis nov spec., a freshwater testate amoeba from Lake Mulan, China

Testate amoebae are an important and abundant component of aquatic ecosystems. Our knowledge of their distribution and the species inventory in Asia is still limited. This paper describes a new planktonic Difflugia from Lake Mulan, province of Hubei, China. These findings are of taxonomic, as well as biogeographic significance because Difflugia mulanensis nov. spec. is a rare and perhaps the only locally distributed member of the well-known urceolate division within this most species-rich genus of lobose testate amoebae. We have studied its morphology and biometry with light and scanning electron microscopical methods. D. mulanensis differs from related species by its very large wide funnel-shaped apertural collar which normally exceeds the width of the test, a relatively long neck, its rounded apical end and the presence of only one nucleus. All shell parameters are normally distributed with low variability. Only the collar height is more variable. This shows that D. mulanensis is a size-monomorphic species. (c) 2005 Elsevier GrnbH. All rights reserved.

Experimental study of trophic cascade effect of silver carp (Hypophthalmichthys molitrixon) in a subtropical lake, Lake Donghu: on plankton community and underlying mechanisms of changes of crustacean community

An enclosure experiment was carried out to test trophic cascade effect of filter-feeding fish on the ecosystem: growth of crustacean zooplankton, and possible mechanism of changes of crustacean community structure. Four fish biomass levels were set as follows: 0, 116, 176 and 316 g m(-2), and lake water ( containing ca. 190 g m(-2) of filter-feeding fishes) was comparatively monitored. Nutrient levels were high in all treatments during the experiment. Lowest algal biomass were measured in fishless treatment. Algal biomass decreased during days 21-56 as a function of fish biomass in treatments of low (LF), medium (MF) and high (HF) fish biomass. Crustaceans biomass decreased with increasing fish biomass. Small-bodied cladocerans, Moina micrura, Diaphanosoma brachyurum and Scapholeberis kingii survived when fish biomass was high whilst, large-bodied cladocerans Daphnia spp. and the cyclopoids Theromcyclops taihokuensis, T. brevifuratus, Mescyclops notius and Cyclops vicinus were abundant only in NF enclosures. Evasive calanoid Sinodiaptomus sarsi was significantly enhanced in LF, but decreased significantly with further increase of fish biomass. Demographic data indicated that M. micrura was well developed in all treatments. Our study indicates that algal biomass might be controlled by silver carp biomass in eutrophic environment. Changes of crustacean community are probably affected by the age of the first generation of species. Species with short generation time were dominant and species with long generation time survived less with high fish biomass. Evasive calanoids hardly developed in treatments with high fish biomass because of the ( bottle neck) effect of nauplii. Species abundance were positively related to fish predation avoidance. Other than direct predation, zooplankton might also be suppressed by filter-feeding fish via competition.

Changes in the plankton community following introduction of filter-feeding planktivorous fish

1. We conducted enclosure experiments in a shallow eutrophic lake, in which a biomass gradient of the filter-feeding planktivore, silver carp, Hypophthalmichthys molitrix Valenciennes, was created, and subsequent community changes in both zooplankton and phytoplankton were examined. 2. During a summer experiment, a bloom of Anabaena flos-aquae developed (approximate to 8000 cells mL(-1)) solely in an enclosure without silver carp. Concurrent with, or slightly preceding the Anabaena bloom, the number of rotifer species and their abundance increased from seven to twelve species (1700-14 400 organisms L-1) after the bloom in this fish-free enclosure. Protozoans and bacteria were generally insensitive to the gradient of silver carp biomass. 3. During an autumn experiment, on the other hand, large herbivorous crustaceans were more efficient than silver carp in suppressing the algae, partly because the lower water temperature (approximate to 24 degrees C) inhibited active feeding of this warm-water fish and also formation of algal colonies. Heterotrophic nanoflagellate and bacterial densities were also influenced negatively by the crustaceans. 4. Correspondence analysis (CA) was applied to the weekly community data of zooplankton and phytoplankton. A major effect detected in the zooplankton community was the presence/absence of silver carp rather than the biomass of silver carp, whereas that in the phytoplankton community was the fish biomass before the Anabaena bloom, but shifted to the presence/absence of the fish after the bloom.

HYDROGRAPHIC VARIATIONS IN SIZE STRUCTURE OF PLANKTON BIOMASS IN A CHANGJIANG FLOODPLAIN LAKE

The size structure of the planktonic community in a Changjiang floodplain lake (Lake Chenhu, Hubei, P. R. China) was described for the inundation period of May through September 1983. The modality of the Sheldon-type size distributions changed hydrographically with the spectral profiles being bimodal during low, rising, mid-high and falling water phases, and trimodal soon after filling and shortly before falling. The modal peaks corresponded respectively to the dominant organisms of chlorophytes and nauplii, while the troughs centered on the bacteria and macrocrustacean size classes in the lake. The slope of the normalized biomass spectrum (an index of plankton size distribution) was less than -1.0 for the filling and falling phases or close to -1.0 for the high water period, indicating that the planktonic biomass tended to decrease or evenly distributes across logarithmically ordered size classes, respectively. This observed variation in the size distribution of the plankton community mainly resulted from changes in water levels and contents of particulate inorganic matter (PIM) in the lake.

PLANKTON AND SESTON STRUCTURE IN A SHALLOW, EUTROPHIC SUBTROPICAL CHINESE LAKE

The seasonal dynamics in the nutrient concentrations, chlorophyll-a amount (Chl-a), total algal volume (CV), Chl-a/CV ratio, seston structure were studied at two sampling stations in a shallow, highly eutrophicated subtropic lake (the Guozheng Hu area of the East Lake) on the plain of the middle basin of the Chang Jiang (the Yangtze River) of China. The lake ecosystem of the Guozheng Hu area is dominated by two planktivorous fishes (silver carp and bighead carp), phytoplankton and zooplankton. Macrophytes are extremely scarce in this area. Concentrations of the total dissolved nitrogen and phosphorus in the Guozheng Hu area in 1990 were very high. Fish yield, of which, more than 90 % was composed of silver carp and bighead carp in the Guozheng Hu area was very high (ca. 1140 kg/ha or 45.6 g/m3 in 1990). Grazing pressure by the fishes on the plankton community is considered to be rather strong. The annual average biomass of zooplankton was ca. 1/3 - 1/2 that of phytoplankton. On the average, dry matter in the living plankton only constituted ca. 3-7 % of the total dry seston, and plankton carbon only constituted ca. 5 - 10 % of the seston carbon. The present results indicate that, in the Guozheng Hu area of the East Lake, of the organic part of the seston, detritus is quantitatively an important constituent, while living plankton is only a very small component.

Exploring factors shaping population genetic structure of the freshwater fish Sinocyclocheilus grahami (Teleostei, Cyprinidae)

Phylogeographical analyses on Sinocyclocheilus grahami samples from seven localities within the Lake Dianchi Basin in China were conducted to explore the main factors shaping population structure within this species. Phylogenetic and network analyses reve

KHAWIA SAUROGOBII N. SP (CESTODA: CARYOPHYLLIDEA) FROM FRESHWATER FISH SAUROGOBIO SPP. (CYPRINIDAE) IN CHINA

Khawia saurogobii n. sp. is described from the intestine of the cyprinid fishes Saurogobio dabryi and Saurogobio dumerili in China. The new species belongs to the Lytocestidae and is placed in Khawia because of its afossate scolex, vitelline follicles in the cortical parenchyma, uterus not looping anterior to the cirrus sac, gonopores separate but close together in distinct genital atrium, external seminal vesicle absent, and postovarian follicles present. The new species differs from other congeneric species by the shape of the body markedly tapering posteriorly from the end of the anterior third, the shape of the scolex that is very short and markedly wider than the neck, spatulate without incisions, but shallow superficial grooves, vitelline follicles and testes starting immediately posterior to the scolex, and an ovary with long, posterior arms bent medially, thus giving the shape of an inverted A.

Impacts of two biomanipulation fishes stocked in a large pen on the plankton abundance and water quality during a period of phytoplankton seasonal succession

Silver and bighead carp were stocked in a large pen to control the nuisance cyanobacterial blooms in Meiliang Bay of Lake Taihu. Plankton abundance and water quality were investigated about once a week from 9 May to 7 July in 2005. Biomass of both total crustacean zooplankton and cladocerans was significantly suppressed by the predation of pen-cultured fishes. There was a significant negative correlation between the N:P weight ratio and phytoplankton biomass. The size-selective predation by the two carps had no effect on the biomass of green alga Ulothrix sp. It may be attributed to the low fish stocking density (less than 40 g m(-3)) before June. When Microcystis dominated in the water of fish pen, the pen-cultured carps effectively suppressed the biomass of Microcystis, as indicated by the significant decline of chlorophyll a in the >38 mu m fractions of the fish pen. Based on the results of our experiment and previous other studies, we conclude that silver and bighead carp are two efficient biomanipulation tools to control cyanobacterial (Microcystis) blooms in the tropical/subtropical eutrophic waters. Moreover, we should maintain an enough stocking density for an effective control of phytoplankton biomass. (C) 2008 Elsevier B.V All rights reserved