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.

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.

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.

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.

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.

Spatial distribution and morphologic diversity of virioplankton in Lake Donghu, China

The spatial distribution and morphological diversity of virioplankton were determined in Lake Donghu which contains three trophic regions: hypertrophic, eutrophic and mesotrophic region. Virioplankton abundance measured by transmission electron microscope (TEM) ranged from 7.7 x 10(8) to 3.0 x 109 ml(-1), being among the highest observed in any natural aquatic system examined so far. The spatial distribution of virioplankton was correlated significantly with chlorophyll a concentration (r = 0.847; P < 0.01) at the sampling sites in Lake Donghu. 76 morphotypes were observed. Most morphotypes have tails, belonging to Siphoviridae, Myoviridae and Podoviridae. The majority of tailed phages in the lake were Myoviridae. Morphotypes which were rarely reported, such as prolate-headed virus-like particles, lemon-shaped virus-like particle, and viruses resembling Tectiviridae and Corticoviridae were all observed in the lake. It is concluded that the high viral abundance might be associated with high density of phytoplankton including algae and cyanobacteria. There was high viral diversity in this eutrophic shallow lake. In addition, cyanophage represented an important fraction of the virioplankton community in Lake Donghu. (c) 2006 Elsevier SAS. All rights reserved.

Variation of benthic diatoms in a shallow, eutrophic Chinese Lake, Lake Donghu

Benthic diatom communities were sampled monthly from May 2004 to May 2005 at four different sites in the littoral zone of Lake Donghu, a shallow eutrophic lake of China. The seasonal patterns of the total abundance, which were lowest in summer and highest in spring, were found at all sites. Total densities of diatom assemblages were significantly higher at hyper-eutrophic sites than at moderately eutrophic sites. Melosira varians was the most abundant species and dominant contributor to total abundance at all sites during spring, autumn and winter, whereas Achnanthes exigua dominated benthic diatom assemblages at the site with the highest nutrient concentrations during the summer. Achnanthes lanceolata var. dubia, Gomphonema parvulum, Navicula similis, Navicula verecunda and Nitzschia amphibia were generally observed at all sites throughout the year and were dominant at higher-nutrient sites. The abundance of ambient nutrients was probably responsible for the spatial variation in biomass, composition and diversity of benthic diatom assemblages, and lake water temperature was the major factor that controlled seasonal distribution.

Sedimentation rates, nitrogen and phosphorus retentions in the largest urban Lake Donghu, China

Sediment core samples were collected in the largest urban Lake Donghu (Stations I and II) in China, and the activities of Pb-210, Ra-226 and Cs-137 were measured by gamma-ray spectrometry. The sedimentation rates, calculated by 210Pb constant rate of supply (CRS) model, ranged from 0.11 to 0.65 (average 0.39) cm(.)y(-1) at Station I, and from 0.21 to 0.78 (average 0.46) cm(.)y(-1) at Station II. Sedimentation rate calculated by Cs-137 as a time marker was 0.55 cm(.)y(-1) at Station II. Based on the average sedimentation rate, we obtained 769 and 147 t(.)y(-1) for nitrogen and phosphorus retentions in Lake Donghu sediments, respectively.

Application of polyurethane foam units and calorimetry to microbial monitoring in Lake Donghu

Combined with the national standard biomonitoring method (polyurethane foam units method), calorimetry was applied to study the metabolic activities of PFU microbial communities in fresh water to determine the effects of anthropotgenic stresses on the activity of the microbial community. Comparisons were made at four sampling stations with different eutrophic status in Lake Donghu. Water quality variables, species number of protozoa, abundances of microorganisms, biomass, heterotrophy indexes and diversity indexes are reported. The heat rate-time curves of the native and concentrated PFU microbial communities were determined at 28 degrees C. Growth rate, measured maximum power output and total heat were calculated from the heat rate-time curves. The values of metabolic variables are higher at the more eutrophic stations, which suggests that organic pollution increases the activity of PFU microbial communities. The metabolic variables are in good agreement with chemical and biotic variables. And calorimetry will be useful for biomonitoring of the PFU microbial community. (C) 2005 Elsevier B.V. All rights reserved.

Detection of extracellular phosphatases in natural spring phytoplankton of a shallow eutrophic lake (Donghu, China)

The species-specific production of extracellular phosphatases in phytoplankton of a subtropical polymictic take was investigated from March to May 2004. Phosphatase activity was detected directly at the site of enzyme action using the enzyme-labelled fluorescence (ELF) technique. Size fractionation of bulk phosphatase activity (PA), concentrations of soluble reactive phosphorus (SRP), chlorophyll a, and phytoplankton composition were determined in parallel. Phosphatase-positive cells were present in every phytoplankton sample; labelled cells were detected in 33 algal taxa, including many chlorophytes, dinoflagellates and some diatoms, but never among cyanobacteria. We recorded an unusual dinoflagellate bloom (Peridiniopsis sp.), of which similar to 25% of the cells were phosphatase-positive. Several populations were partly phosphatase-positive whenever present, while some other species never showed any activity. The production of extracellular phosphatases was not primarily regulated by ambient P concentrations; algae produced these enzymes even if SRP concentrations were high. Moreover, heterotrophic nanoflagellates most probably contributed to the pool of particle-bound PA in some samples.

Attenuation of photosynthetically available radiation by chlorophyll, chromophoric dissolved organic matter, and tripton in lake Donghu, China

The attenuation coefficient of photosynthetically available radiation [K-d(PAR)] and three water quality parameters [chlorophyll a (chl a)], chromophoric dissolved organic matter (CDOM) and tripton] were measured at three stations in shallow, subtropical Lake Donghu from April 2003 to March 2004. The multiple regression equation of K-d(PAR) versus chl a, CDOM, and tripton was: K-d(PAR) = 0.44 + 0.019 chl a + 1.88 CDOM + 0.016 tripton, which revealed the relative contributions of the three parameters to K-d(PAR). The effects of water and CDOM on K-d(PAR) were of minor importance (19-26%), while chl a and tripton were the two greatest contributors, accounting collectively for 74-81%.

Changes in the Phytoplankton community of Lake Donghu since the 1980s

The phytoplankton of Lake Donghu was compared between two periods (i.e., 1982-83 and 1999-2001). During 1982-1983, Lake Donghu was characterized by cyanobacterial blooms, very high summer phytoplankton biomass, and a spring clear-water phase. However, during 1999-2001, the lake was dominated by dinophytes and diatoms and had a relatively low phytoplankton biomass and a very turbid state.. Increased stocking of silver carp and bighead carp during the interim years eliminated cyanobacterial blooms and large-bodied daphnids from the lake, creating favorable conditions for growth of the small-sized algae.

Nonylphenol, bisphenol-A and DDTs in Lake Donghu, China

A preliminary survey was carried out in April 2003 to estimate the levels of nonylphenol (NP), bisphenol-A (BPA) and residual DDTs in Lake Donghu. After then, the sediments of some areas of the lake were moved out for clearing the lake by the local government. Therefore, the variance of NP and BPA after the clearance of sediment in the surface water was determined from December 2003 to May 2004. Sediments, surface water and fish were collected from four sub-lake areas and the analytes were qualified by GC/MS in SIM mode after concentration onto an Oasis solid phase extraction cartridge. NP and BPA values ranged between 5.46-119.10 and 0.9913.42 mg/kg dw, respectively, in sediments, 75.2-179.6 and 15.1-62.5 mu g/L in surface water. Meanwhile, the bioaccumulation factors (BCFs) of NP and BPA were calculated in fish livers. Plasma vitellogenin (VTG), a sensitive biomarker showing oestrogenicity was detected in the captured male Wuchang bream (Megalobrama amblycephala Yih) and common carp (Cyprinus carpio). After sediment clearance, the concentrations of NP and BPA decreased to 0.65-25.04 and 0.04-21.32 mu g/L. The results indicate the presence of high-dose xenoestrogenic NP and BPA, probably major contributors which associate with VTG induction in Lake Donghu male fish.

Studies on the food web structure of Lake Donghu using stable carbon and nitrogen isotope ratios

Food web structure was studied by using carbon and nitrogen isotope ratios in a hypereutrophic subtropical Chinese lake, Lake Donghu. High external nutrient loading and the presence of abundant detritus from submersed macrophytes were responsible for the high sediment delta(15)N and delta(13)C, respectively. C-13 was significantly higher in submersed macrophytes than in other macrophytes. The similar delta(13)C values in phytoplankton, zooplankton, zoobenthos, and planktivorous fish indicate that phytoplankton was the major food source for the consumers. By using a delta(15)N mass balance model, we estimate that the contributions of zooplankton to the diet of silver carp and bighead carp were 54% and 74%, respectively, which is in agreement with previous microscopic observations on intestinal contents of these fishes.

Growth and production of free-living heterotrophic nanoflagellates in a eutrophic lake - Lake Donghu, Wuhan, China

In situ growth of heterotrophic nanoflagellates (HNF) in Lake Donghu, a eutrophic shallow lake in mainland China, was studied from January 1999 to March 2000 using a modified Weisse protocol. The study results indicated that the growth rates of HNF showed pronounced seasonal variation (-0.37-1.25 d(-1)), reaching the maximum during spring to early summer. When the water temperature was higher than 25.5 degreesC, HNF growth was inversely proportional to water temperature. There was an effect by bacterial abundance and autotrophic picoplankton on HNF growth that depended on location. HNF biomass was the highest in late spring, and the HNF production ranged from -2.25 to 35.45 mg l(-1) d(-1) with mean of 3.17 mg l(-1) d(-1). When considered in the context of biomass and production data for zooplankton in Lake Donghu, it was evident that HNF contributed significantly to the total zooplankton production in Lake Donghu. These in situ studies indicate that temperature and food supply are the major determinants of HNF abundance and productivity.