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.

Characteristics of the microbial communities in the integrated vertical-flow constructed wetlands

Microorganisms play an important role in removing pollutants from constructed wetlands. We investigated the microbial characteristics in a novel integrated vertical-flow constructed wetland (IVCW), which has been in operation in Wuhan, China since 1998. We used phospholipid fatty acid (PLFA) and amoA gene to analyze the structure and diversity of the microbial community within the IVCW. PLFA results suggested that the amount of bacterial PLFA was significantly higher than that of fungal PLFA, but the total microbial biomass represented by PLFA index was low in the system. Microbial spatial distribution showed significantly higher bacterial (both G(+) and G(-)) and fungal biomass in the surface than in the subsurface layers. The ratios of monounsaturated to branched PLFA demonstrated that an anaerobic layer sandwiched by two aerobic layers existed in the IVCW, consistent with the redox potential results. Analysis of the amoA revealed the presence of Nitrosomonas-like sequences in the surface substrate of the downflow chamber and apparent diversities of ammonia-oxidizing bacteria in the system. These results suggest that microorganisms, despite their relatively low biomass, have inhabited the IVCW, and the results will offer some valuable information on microbe to system designers and managers.

Communities of gastrointestinal helminths of fish in historically connected habitats: habitat fragmentation effect in a carnivorous catfish Pelteobagrus fulvidraco from seven lakes in flood plain of the Yangtze River, China

Background: Habitat fragmentation may result in the reduction of diversity of parasite communities by affecting population size and dispersal pattern of species. In the flood plain of the Yangtze River in China, many lakes, which were once connected with the river, have become isolated since the 1950s from the river by the construction of dams and sluices, with many larger lakes subdivided into smaller ones by road embankments. These artificial barriers have inevitably obstructed the migration of fish between the river and lakes and also among lakes. In this study, the gastrointestinal helminth communities were investigated in a carnivorous fish, the yellowhead catfish Pelteobagrus fulvidraco, from two connected and five isolated lakes in the flood plain in order to detect the effect of lake fragmentation on the parasite communities. Results: A total of 11 species of helminths were recorded in the stomach and intestine of P. fulvidraco from seven lakes, including two lakes connected with the Yangtze River, i.e. Poyang and Dongting lakes, and five isolated lakes, i.e. Honghu, Liangzi, Tangxun, Niushan and Baoan lakes. Mean helminth individuals and diversity of helminth communities in Honghu and Dongting lakes was lower than in the other five lakes. The nematode Procamallanus fulvidraconis was the dominant species of communities in all the seven lakes. No significant difference in the Shannon-Wiener index was detected between connected lakes (0.48) and isolated lakes (0.50). The similarity of helminth communities between Niushan and Baoan lakes was the highest (0.6708), and the lowest was between Tangxun and Dongting lakes (0.1807). The similarity was low between Dongting and the other lakes, and the similarity decreased with the geographic distance among these lakes. The helminth community in one connected lake, Poyang Lake was clustered with isolated lakes, but the community in Dongting Lake was separated in the tree. Conclusion: The similarity in the helminth communities of this fish in the flood-plain lakes may be attributed to the historical connection of these habitats and to the completion of the life-cycles of this fish as well as the helminth species within the investigated habitats. The diversity and the digenean majority in the helminth communities can be related to the diet of this fish, and to the lacustrine and macrophytic characters of the habitats. The lake isolation from the river had little detectable effect on the helminth communities of the catfish in flood-plain lakes of the Yangtze River. The low similarities in helminth communities between the Dongting Lake and others may just be a reflection of its unique water environment and anthropogenic alterations or fragmentation in this lake.

Changes in benthic algal communities following construction of a run-of-river dam

Ecological responses to dam construction are poorly understood, especially for downstream benthic algal communities. We examined the responses of benthic algal communities in downstream reaches of a tributary of the Xiangxi River, China, to the construction of a small run-of-river dam. From February 2003 to August 2006, benthic algae, chemical factors, and habitat characteristics were monitored upstream and downstream of the dam site. This period spanned 6 mo before dam construction and 37 mo after dam construction. Benthic algal sampling yielded 199 taxa in 59 genera that belonged to Bacillariophyta, Chlorophyta, and Cyanophyta. Some physical factors (flow velocity, water depth, and channel width) and 3 algal metrics (diatom species richness, Margalef diversity, and % erect individuals) were significantly affected by the dam construction, whereas chemical factors (e.g., NH4-N, total N, SiO2) were not. Nonmetric multidimensional scaling (NMS) ordinations showed that overall algal assemblage structure downstream of the dam sites was similar to that of upstream control sites before dam construction and for 1 year after dam construction (p > 0.05). However, sites belonging to upstream and downstream reaches were well separated on NMS axis 1 during the 2(nd) and 3(rd) years after dam construction. Our results suggest that impacts of dam construction on benthic algal communities took 2 to 3 y to emerge. Further development of a complete set of indicators is needed to address the impact of small-dam construction. Our observations underscore the need for additional studies that quantify ecological responses to dam construction over longer time spans.

Ecology of macrozoobenthic communities in two plateau lakes of Southwest China

Ecological studies on macrozoobenthos were conducted in two small plateau lakes in the Yunnan-Guizhou Plateau, Southwest China: Xingyun Lake (XL), a eutrophic lake whose main source of primary production was phytoplankton (Chl a=99.76 +/- 24.01 mu g/L), and Yangzong Lake (YL), a mesotrophic lake. Sampling was carried out from October 2002 to May 2004. Altogether 23 benthic taxa were identified in XL and 21 taxa in YL. The density of benthos in XL was much lower than that in YL, but the biomass was about equal in the two lakes, being 1 423 ind/m(2) and 8.71 g/m(2) in XL and 4 249 ind/m(2) and 8.60 g/m(2) in YL. The dominant species were Limnodrilus hoffmeisteri, Branchiura sowerbyi, Aulodrilus pluriseta and Chironomus sp. in XL and Limnodrilus hoffmeisteri, Aulodrilus pluriseta and Bellamya sp. in YL. Seasonal fluctuation occurred, showing richer species in summer and winter, but the density and biomass varied in different ways in the two lakes. Analyses on functional feeding groups indicate that collector-gatherers were predominant, but the relative abundances of other groups were different. Stepwise multiple regression analysis demonstrated that the water depth, conductivity and chlorophyll a were the key factors affecting macrozoobenthic abundance in the lakes.

Response of macrozoobenthos communities to ecological engineering remediation in a hypertrophic urban lake

The relationship of macrozoobenthos communities with some environmental variables, and their response to the ongoing restoration measures were studied in a small hypertrophic urban lake near the Yangzte River, China. Twenty taxa including 9 oligochaetes, 7 insects, 2 mollusks and two other animals were found during March 2005 to May 2006. The reappearance of some indigenous macrozoobenthos species showed that the ecological engineering remediation carried out was helpful for the recovery of the macrozoobenthos communities. Through canonical correspondence analysis (CCA), it was detected that temperature (T), conductivity (COND), Secchi depth/deep (SD/Deep), TN, total suspended solids (SS) and chemical oxygen demand (CODcr) were significant environmental factors that influenced the pattern of macrozoobenthos. Limnodrilus hoffmeisteri, Tanypus sp. and Alocinma longicornis could be used as potential bio-indicators in monitoring the development of ecological restoration.

Seasonal patterns of sedimentation and their associations with benthic communities in Xiangxi Bay of the Three Gorges Reservoir, China

Sedimentation variables and benthic community data were collected at seven stations during four seasons in Xiangxi Bay of the Three Gorges Reservoir, China. Summer, the season of highest discharge into the reservoir, was characterized by the extreme sediment loading. The benthic macroinvertebrate community was dominated by oligochaetes across all seasons at most stations. In winter/spring, macroinvertebrate density and richness increased. Correspondence analysis showed that community structure differed among stations at the two ends of the bay in winter and among almost all stations in spring, However, no variable associated with sedimentation appeared to be associated with differences in the community.

Spatiotemporal heterogeneity of plankton communities in Lake Donghu, China, as revealed by PCR-denaturing gradient gel electrophoresis and its relation to biotic and abiotic factors

The 16S and 18S rRNA genes of planktonic organisms derived from five stations with nutrient gradients in Lake Donghu, China, were studied by PCR-denaturing gradient gel electrophoresis (DGGE) fingerprinting, and the relationships between the genetic diversity of the plankton community and biotic/abiotic factors are discussed. The concentrations of total nitrogen (TN), total phosphorus (TP), NH4-N and As were found to be significantly related (P < 0.05) to morphological composition of the plankton community. Both chemical and morphological analyses suggested that temporal heterogeneity was comparatively higher than spatial heterogeneity in Lake Donghu. Although the morphological composition was not identical to the DGGE fingerprints in characterizing habitat similarity, the two strongest eutrophic stations (I and II) were always initially grouped into one cluster. Canonical correspondence analysis suggested that the factors strongly correlated with the first two ordination axes were seasonally different. The concentrations of TN and TP and the densities of rotifers and crustaceans were generally the main factors related to the DGGE patterns of the plankton communities. The study suggested that genetic diversity as depicted by metagenomic techniques (such as PCR-DGGE fingerprinting) is a promising tool for ecological study of plankton communities and that such techniques are likely to play an increasingly important role in assessing the environmental conditions of aquatic habitats.

Monitoring bioaccumulation and toxic effects of hexachlorobenzene using the polyurethane foam unit method in the microbial communities of the Fuhe River, Wuhan

Hexachlorobenzene (HCB) is a chlorinated aromatic hydrocarbon that was widely used for seed dressing in prevention of fungal growth on crops, and also as a component of fireworks, ammunition, and synthetic rubbers. Because of its resistance to degradation and mobility, HCB is widely distributed throughout the environment and is accumulated through food chains in different ecosystems. In this study, a preliminary investigation was carried out on the bioaccumulation and the toxic effects of HCB in the microbial (protozoan in particular) communities in the Fuhe River, Wuhan, a water body receiving industrial wastewaters containing HCB and other pollutants, using the standardized polyurethane foam units (PFU) method. Field samples were taken from eight stations established along the Fuhe River in January and August 2006. The concentration ratios of HCB in microbial communities and in water were 9.66-18.64, and the microbial communities accumulated 13.29-56.88 mu g/L of HCB in January and 0.82-10.25 mu g/L HCB in August. Correlation analysis showed a negative correlation between the HCB contents in the microbial assemblage, and the number of species and the diversity index of the protozoan communities. This study demonstrated the applicability of the PFU method in monitoring the effects of HCB on the level of microbial communities.

Studies on the restoration succession of PFU microbial communities in a pilot-scale microcosm

In order to imitate the restoration succession process of natural water ecosystem, a laboratory microcosm system of constant-flow-restoration was designed and established. A eutrophycation lake, Lake Donghu, was selected as the subject investigated. Six sampling stations were set on the lake, among which the water of station IV was natural clean water, and others were polluted with different degrees. Polyurethane foam unit microbial communities, which had colonized in the stations for a month, were collected from these stations and placed in their respective microcosms, using clean water of station IV to gradually replace the water of these microcosms. In this process, the healthy community in clean water continuously replaced the damaged communities in polluted water, the restoration succession of the damaged communities was characterized by weekly determination of several functional and structural community parameters, including species number (S), diversity index (DI), community pollution value (CPV), heterotrophy index (HI), and similarity coefficient. Cluster analysis based on similarity coefficient was used to compare the succession discrepancies of these microbial communities from different stations. The ecological succession of microbial communities during restoration was investigated by the variable patterns of these parameters, and based on which, the restoration standards of these polluted stations were suggested in an ecological sense. That was, while being restored, the water of station 0 (supereutrophycation) should be substituted with natural clean water by 95%; station I (eutrophycation), more than 90%; station II (eutrophycation), more than 85%; station III (eutrophycation), about 85%; station V (mesoetitrophycation), less than 50%. The effects of the structural and functional parameters in monitoring and assessing ecological restoration are analyzed and compared. (C) 2007 Elsevier Ltd. All rights reserved.

Development of the microbial communities in Lake Donghu in relation to water quality

There is increasing recognition that protozoa is very useful in monitoring and evaluating water ecological healthy and quality. In order to study the relationship between structure and function of protozoan communities and water qualities, six sampling stations were set on Lake Donghu, a hypereutrophic subtropical Chinese lake. Microbial communities and protists sampling from the six stations was conducted by PFU (Polyurethane foam unit) method. Species number (S), diversity index (DI), percentage of phytomastigophra, community pollution value (CPV), community similarity and heterophy index (HI) were mensurated. The measured indicators of water quality included total phosphorus (TP), dissolved oxygen (DO), Chemical oxygen demand (COD), NH4 (+), NO2 (-) and NO3-. Every month water samples from stations I, II, III, IV were chemically analyzed for a whole year, Among the chemically analyzed stations, station I was the most heavily polluted, station II was the next, stations III and IV had similar pollution degrees. The variable tendencies of COD, TP, NH3, NO2-, NO3-, and DO during the year was approximately coincident among the six stations. Analysis from the community parameters showed that the pollution of station 0 was much more serious than others, and station V was the most slight. Of the community parameters, CPV and HI were sensitive in reflecting the variables of the water quality. Community similarity index was also sensitive in dividing water qualities and the water quality status of different stations could be correctly classified by the cluster analysis. DI could reflect the tendency of water quality gradient, species number and percentage of Phytomastigophora was not obvious in indicating the water quality gradient.

Effect of plant architecture on the structure of epiphytic macro invertebrate communities in a Chinese lake

We examined the effect of different plant architecture types on epiphytic macroinvertebrates of a shallow macrophyte-dominated lake in China. Macroinvertebrates were sampled from four dominant submersed macrophytes in the lake - two dissected plants (Myriophyllum spicatum L. and Ceratophyllum demersum L.) and two undissected plants (Potamogeton maackianus A. Benn. and Vallisneria spiralis L.). Macro invertebrate richness showed significant differences among four submersed macrophyte habitats, and higher density per g of dry plant were associated with dissected plants than undissected plants. The average abundance in dissected plants was as three-six times as in undissected plants. The biodiversity of epiphytic macroinvertebrates was higher in dissected plants than undissected plants. Our results suggest that dissected plants provide different habitat for macroinvertebrates than dissected plant, and this concurs with the hypothesis that the former could support more epiphytic macroinvertebrates than the latter.

Using epilithic diatom communities to assess ecological condition of Xiangxi River system

Assessment method for ecological condition of Xiangxi River system was studied by using 13 candidate metrics of epilithic diatom which can reflect conditions in pH, salinity, nitrogen uptake metabolism, oxygen requirements, saprobity, trophic state, morphological character and pollution tolerant capability etc. By one-way ANOVA, the metrics of relative abundance of acidobiontic algae (ACID), freshwater algae (FRESH), high oxygen requirement (HIGH-O), eutraphentic state (EUTRA) and mobile taxa ( MOBILE) were suitable for distinguishing sites in different conditions. Then, the river diatom index (RDI) composed of these five metrics was used to evaluate ecological condition of the river. The results showed that the healthiest sites were in the Guanmenshan Natural Reserve ( with the mean RDI of 79.73). The sites located in tributary of Jiuchong River also owned excellent state (mean RDI of 78.25). Mean RDI of another tributary - Gufu River and the main river were 70.85 and 68.45 respectively, and the unhealthiest tributary was Gaolan River (with mean RDI of 65.64). The mean RDI for all the 51 sites was 71.40. The competence of RDI was discussed with comparison of evaluation results of DAIpo and TDI, it can be concluded that multimetrics is more competent in assessment task.

Monitoring organochlorine pesticides and polychlorinated biphenyls in Baiyangdian Lake using microbial communities

We used polyurethane foam units (PFUs) to collect persistent organic pollutants (POPs) from four sites in Baiyangdian Lake in July 2003. Following extraction from the PFUs, relative concentrations of seven organochlorine pesticides (OCPs) and ten polychlorinated biphenyls (PCBs) were determined by gas chromatography. OCPs and PCBs were detected in the microbial communities from all the four sampling stations. In terms of the total concentration of POPs (OCPs+ PCBs), two river estuary stations had more POP (18.45 mu g/L and 9.77 mu g/L) than the two mid-lake stations (4.75 mu g/L and 5.21 mu g/L), indicating that Baiyangdian Lake was significantly impacted by inflow from the Fu River and Baigou River.

Distribution of the macroinvertebrate communities in the Xiangxi River system and relationships with environmental factors

Surveys of macroinvertebrates were carried out in the Xiangxi River system during July of 2001. Among the 121 taxa collected, Ephemeroptera, Trichoptera, and Diptera dominated (41.7, 26.0, and 24.5% of the total relative abundance, respectively). Two-way indictor species analysis and detrended correspondence analysis divided the 49 sites into four groups based on species composition and relative abundance. Canonical correspondence analysis indicated that elevation, SiO2, pH, conductivity, hardness, and NO2-N were significant environmental factors affecting the distribution of macroinvertebrates.