Nearly a decade-long repeatable seasonal diversity patterns of bacterioplankton communities in the eutrophic Lake Donghu (Wuhan, China).

Uncovering which environmental factors govern community diversity patterns and how ecological processes drive community turnover are key questions related to understand the community assembly. However, the ecological mechanisms regulating long-term variations of bacterioplankton communities in lake ecosystems remain poorly understood. Here we present nearly a decade-long study of bacterioplankton communities from the eutrophic Lake Donghu (Wuhan, China) using 16S rRNA gene amplicon sequencing with MiSeq platform. We found strong repeatable seasonal diversity patterns in terms of both common (detected in more than 50% samples) and dominant (relative abundance >1%) bacterial taxa turnover. Moreover, community composition tracked the seasonal temperature gradient, indicating that temperature is a key environmental factor controlling observed diversity patterns. Total phosphorus also contributed significantly to the seasonal shifts in bacterioplankton composition. However, any spatial pattern of bacterioplankton communities across the main lake areas within season was overwhelmed by their temporal variabilities. Phylogenetic analysis further indicated that 75%-82% of community turnover was governed by homogeneous selection due to consistent environmental conditions within seasons, suggesting that the microbial communities in Lake Donghu are mainly controlled by niche-based processes. Therefore, dominant niches available within seasons might be occupied by similar combinations of bacterial taxa with modest dispersal rates throughout different lake areas.

Alpha-tubulin and small subunit rRNA phylogenies of peritrichs are congruent and do not support the clustering of mobilids and sessilids (Ciliophora, Oligohymenophorea).

Peritrich ciliates have been traditionally subdivided into two orders, Sessilida and Mobilida within the subclass Peritrichia. However, all the existing small subunit (SSU) rRNA phylogenetic trees showed that the sessilids and mobilids did not branch together. To shed some light on this disagreement, we tested whether or not the classic Peritrichia is a monophyletic group by assessing the reliability of the SSU rRNA phylogeny in terms of congruency with alpha-tubulin phylogeny. For this purpose, we obtained 10 partial alpha-tubulin sequences from peritrichs and built phylogenetic trees based on alpha-tubulin nucleotide and amino acid data. A phylogenetic tree from the alpha-tubulin and SSU rRNA genes in combination was also constructed and compared with that from the SSU rRNA gene using a similar species sampling. Our results show that the mobilids and sessilids are consistently separated in all trees, which reinforces the idea that the peritrichs do not constitute a monophyletic group. However, in all alpha-tubulin gene trees, the urceolariids and trichodiniids do not group together, suggested mobilids may not be a monophyletic group.

Impacts of algal blooms removal by chitosan-modified soils on zooplankton community in Taihu Lake, China.

It is important to assess the effect on zooplankton when perform the environmental protection or restoration technology, especially removing algal blooms, because algae were the major primary producer in algal lakes. The influence on zooplankton community after half a year of algal blooms removed by chitosan-modified soils in Taihu Lake was assessed and the rationality of carrying out the process semiannually was evaluated in the present study. Morphological composition and genetic diversity of zooplankton community were investigated by microscope checkup and polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE). A total of 44 zooplankton taxa (23 protozoa, 17 rotifers, 3 copepoda and 1 cladocera) were detected by microscope checkup, and a total of 91 bands (28 bands amplified by primers F1427-GC and R1616, 63 bands amplified by primers Fung-GC and NS1) were detected by PCR-DGGE. The results of cluster analysis or detrended correspondence analysis indicated that there was no considerable difference in morphological composition of zooplankton and DGGE profiles between experimental and control sites, and DGGE profiles could represent the biologic diversity. The study showed that zooplankton community could recover original condition after half year of algal blooms removed by chitosan-modified soils and it was acceptable to apply this process semiannually. In addition, the results revealed that PCR-DGGE could be applied to investigate the impacts of the environmental protection or restoration engineering on zooplankton community diversity.

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.

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), NH(4)-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.

Plankton community composition in the Three Gorges Reservoir Region revealed by PCR-dGGE and its relationships with environmental factors.

To explore the relationships between community composition and the environment in a reservoir ecosystem, plankton communities from the Three Gorges Reservoir Region were studied by PCR-denaturing gradient gel electrophoresis fingerprinting. Bacterial and eukaryotic operational taxonomic units (OTUs), generated by DGGE analysis of the PCR-amplified 16S and 18S rRNA genes, were used as surrogates for the dominant "biodiversity units". OTU composition among the sites was heterogeneous; 46.7% of the total bacterial OTUs (45) and 64.1% of the eukaryotic OTUs (39) were identified in less than half of the sampling sites. Unweighted pair group method with arithmetic averages (UPGMA) clustering of the OTUs suggested that the plankton communities in the Xiangxi River sites were not always significantly different from those from the Yangtze River sites, despite clear differences in their environmental characterizations. Canonical correspondence analysis (CCA) was applied to further investigate the relationships between OTU composition and the environmental factors. The first two CCA ordination axes suggested that the bacterial community composition was primarily correlated with the variables of NO(3-)-N, dissolved oxygen (DO), and SiO3(2-)-Si, whereas, the eukaryotic community was mainly correlated with the concentrations of DO, PO4(3-)-P, and SiO3(2-)-Si.

Monitoring of organochlorine pesticides using PFU systems in Yunnan lakes and rivers, China.

Polyurethane foam unit (PFU) systems were collected from 11 lakes and three rivers in the Yunnan Plateau, China and, the PFU extrusion liquids were analyzed for organochlorine pesticides (OCPs) by gas chromatography with electron capture detection (GC-ECD). The concentrations of pp'-DDE, HCB and HCHs were undetectable to 1.86 microgl-1 (mean 0.27 microgl-1), undetectable to 0.72 microgl-1 (mean 0.11 microgl-1), and 0.24-21.95 microgl-1 (mean 7.39 microgl-1) respectively in lakes; and those in rivers were undetectable to 0.23 microgl-1 (mean 0.08 microgl-1), 0.68-2.93 microgl-1 (mean 1.70 microgl-1), and 2.71-37.56 microgl-1 (mean 17.01 microgl-1) respectively. Notably, some residue levels of OCPs exceeded the US National Recommended Water Quality Criteria, implying Yunnan has levels of OCPs potentially harmful to human health. Further, the contamination by OCPs showed an obvious spatial distribution pattern. Amongst the lakes, Dianchi, Xingyun, Lugu and Yangzonghai had the highest OCP levels dominated by beta-HCH, whereas among rivers, Nujiang and Lancang Rivers had the highest contents of OCPs dominated by alpha-HCH. This demonstrates that HCHs are the predominant contaminants and some point sources of HCHs may still exist in Yunnan. The pollution levels in Yunnan were compared with other studies, suggesting the PFU method is suitable for long-term on-line monitoring of trace OCPs in aquatic ecosystems. Therefore, continuous studies monitoring OCPs in lakes and rivers are needed to further understand the future trend of contamination.

Effects of high ammonia concentrations on three cyprinid fish: Acute and whole-ecosystem chronic tests.

A number of studies have revealed ammonia to be toxic to aquatic organisms; however, little is known about its effects under natural conditions. To elucidate the role of ammonia, we conducted 96-h acute toxicity tests as well as a whole-ecosystem chronic toxicity test for one year in ten 600-m2 ponds. Three common cyprinids, silver carp Hypophthalmichthys molitrix Val. (H.m.), bighead carp Aristichthys nobilis Richardson (A.n.), and gibel carp Carassius auratus gibelio Bloch (C.g.), were used as test organisms. The 96-h LC50 values of un-ionized ammonia (NH3) for H.m., A.n., and C.g. were 0.35, 0.33, and 0.73mgL-1, respectively. In the ponds, annual mean NH3 ranged between 0.01 and 0.54mgL-1, with 4 ponds having a NH3 higher than the LC50 of A.n. (lowest LC50 in this study). No fish were found dead in the high-nitrogen ponds, but marked histological changes were found in livers and gills. Despite these changes, the specific growth rate of H.m. and A.n. increased significantly with NH3. Our pond results suggest that fish might be more tolerant to high ammonia concentrations in natural aquatic ecosystems than under laboratory conditions. Our finding from field experiments thus suggests that the existing regulatory limits for reactive nitrogen (NH3) established from lab toxicity tests might be somewhat too high at the ecosystem conditions. Field-scale chronic toxicity tests covering full life histories of fish and other aquatic organisms are therefore encouraged in order to optimize determination of the effects of ammonia in natural environments.

Host species as a strong determinant of the intestinal microbiota of fish larvae.

We investigated the influence of host species on intestinal microbiota by comparing the gut bacterial community structure of four cohabitating freshwater fish larvae, silver carp, grass carp, bighead carp, and blunt snout bream, using denaturing gradient gel electrophoresis (DGGE) of the amplified 16S and 18S rRNA genes. Similarity clustering indicated that the intestinal microbiota derived from these four fish species could be divided into four groups based on 16S rRNA gene similarity, whereas the eukaryotic 18S rRNA genes showed no distinct groups. The water sample from the shared environment contained microbiota of an independent group as indicated by both 16S and 18S rRNA genes segments. The bacterial community structures were visualized using rank-abundance plots fitted with linear regression models. Results showed that the intestinal bacterial evenness was significantly different between species (P<0.05) and between species and the water sample (P<0.01). Thirty-five relatively dominant bands in DGGE patterns were sequenced and grouped into five major taxa: Proteobacteria (26), Actinobacteria (5), Bacteroidetes (1), Firmicutes (2), and Cyanobacterial (1). Six eukaryotes were detected by sequencing 18S rRNA genes segments. The present study suggests that the intestines of the four fish larvae, although reared in the same environment, contained distinct bacterial populations, while intestinal eukaryotic microorganisms were almost identical.

[Community structure characteristics of zooplankton in Yili region of Xinjiang in summer].

In June-July 2006, investigations were made on the zooplankton in the mainstream of Yili River and its three main tributaries (Tekes River, Künes River and Kax River), two reservoirs (Qiapuqihai Reservoir and Jilintai Reservoir), intermittent waters, and ponds in Yili region of Xin-jiang. A total of 217 species belonging to 134 genera were identified, among which, Protozoa and Rotifer were the main groups, which contained 153 species (71% of the total species number) of 95 genera and 51 species (23% of the total) of 27 genera, respectively. Only 8 species of Cladocera and 5 species of Copepoda were observed. The zooplankton richness ranged from 7 to 132, with the lowest (7) in Jilintai Reservoir, and the highest (132) in Yili River. Qiapuqihai Reservoir had the highest density and biomass of zooplankton (11391 ind x L(-1) and 2.79 mg x L(-1), respectively), but Tekes River had the lowest density (578 ind x L(-1)) and Kax River had the lowest biomass of zooplankton (0.03 mg x L(-1)). Pearson correlation analysis and linear regressions showed a significantly negative relationship between zooplankton species number and water flow velocity. Based on the species number, density, and biomass of zooplankton, it was found that Yili River and its three main tributaries were in oligotrophic status, while Qiapuqihai Reservoir and Jilintai Reservoir were in mesotrophic status.

Molecular phylogeny of Stentor (Ciliophora: Heterotrichea) based on small subunit ribosomal RNA sequences.

To determine the phylogenetic position of Stentor within the Class Heterotrichea, the complete small subunit rRNA genes of three Stentor species, namely Stentor polymorphus, Stentor coeruleus, and Stentor roeseli, were sequenced and used to construct phylogenetic trees using the maximum parsimony, neighbor joining, and Bayesian analysis. With all phylogenetic methods, the genus Stentor was monophyletic, with S. roeseli branching basally.

[DNA fingerprinting structure of plankton community and its relations to environmental physical-chemical factors in Donghu Lake].

By the method of RAPD fingerprinting, this paper studied the DNA fingerprinting structure of plankton community and its relations to the main environmental physical-chemical factors at five sites in Donghu Lake. From the screened 9 random primers, a total of 210 observable bands with a length of 150-2 000 bp were amplified, 93.3% of which were polymorphic. At the five sites, the average number of amplified bands was 42, with the maximum (53) at site IV and the minimum (35) at site V. The PO4(3-)-P and TP contents were the highest at site I, NH4(+)-N, TN and NO2(-)-N contents were the highest at site V, while the values of all test physical-chemical parameters were the lowest at site IV. No obvious differences in COD, alkalinity, rigidity, and calcium content were observed among the study sites. Similarity clustering analysis showed that the DNA fingerprinting of plankton community based on RAPD marker could cluster the five sites into two groups, i. e., sites I, II and III could be clustered into one group, while sites IV and V could be clustered into another group, which was consistent with the clustering analysis based on the main environmental physical-chemical factors. In conclusion, there was a close relation between the DNA fingerprinting structure of plankton community and the main environmental physical-chemical factors in Donghu Lake.

Use of PFU protozoan community structural and functional characteristics in assessment of water quality in a large, highly polluted freshwater lake in China.

Structural and functional parameters of protozoan communities colonizing on PFU (polyurethane foam unit) artificial substrate were assessed as indicators of water quality in the Chaohu Lake, a large, shallow and highly polluted freshwater lake in China. Protozoan communities were sampled 1, 3, 6, 9 and 14 days after exposure of PFU artificial substrate in the lake during October 2003. Four study stations with the different water quality gradient changes along the lake were distinguishable in terms of differences in the community's structural (species richness, individual abundance, etc.) and functional parameters (protozoan colonization rates on PFU). The concentrations of TP, TN, COD and BOD as the main chemical indicators of pollution at the four sampling sites were also obtained each year during 2002-2003 for comparison with biological parameters. The results showed that the species richness and PFU colonization rate decreased as pollution intensity increased and that the Margalef diversity index values calculated at four sampling sites also related to water quality. The three functional parameters based on the PFU colonization process, that is, S(eq), G and T90%, were strongly related to the pollution status of the water. The number of protozoan species colonizing on PFU after exposure of 1 to 3 days was found to give a clear comparative indication of the water quality at the four sampling stations. The research provides further evidence that the protozoan community may be utilized effectively in the assessment of water quality and that the PFU method furnishes rapid, cost-effective and reliable information that may be useful for measuring responses to pollution stress in aquatic ecosystems.