The role of cell-surface-bound phosphatases in species competition within natural phytoplankton assemblage: an in situ experiment

Despite it is widely acknowledged that the ability to hydrolyze dissolved organic matter using extracellular phosphatases is diverse in fresh water phytoplankton, the competition within single species related to presence and quantity of cell-surface-bound phosphatases has not been examined in natural conditions yet. Here, we studied phytoplankton species competition in a freshwater reservoir during an in situ experiment. A natural plankton community, with the exclusion of large zooplankton, was enclosed in permeable dialysis bags inside two large containers of different bioavailable phosphate concentrations. Phytoplankton species biomass and the abundance of bacteria were determined in purpose to compare the development of enclosed microbial communities. Total and cell-surface-bound phosphatase activities in the phytoplankton were investigated using the Fluorescently Labelled Enzyme Activity (FLEA) technique that allows for direct microscopic detection of phosphatase-positive cells and, with image cytometry, enables quantification of phosphatase hydrolytic capacity. Production of extracellular phosphatases was not completely inhibited or stopped in the phosphate-enriched environment, phytoplankton cells only showed the activity less often. Under the phosphate-nonenriched conditions, the production of phosphatases was enhanced, but active species did not proliferate amongst phytoplankton assemblage. Further, specific growth rates of the phosphatase-positive species in the non-enriched environment were lower than the same phosphatase-positive species in phosphate-enriched environment. Interestingly, the phosphatase-positive cells of Ankyra ancora increased their size in both treatments equally, although the population in phosphate-enriched environment grew much faster and the cell-specific phosphatase activity was lower. We hypothesize that brand new daughter cells had sufficient phosphorus reserves and therefore did not employ extracellular phosphatases until they matured and needed extra bioavailable phosphorus to support their metabolism before cell division. Based on presented in situ experiment, we propose that the ability to hydrolyze organic polymers and particles with cell-surface-hound phosphatases is advantageous for longer persistence of given population in a phosphate-scarce environment; although phosphatase-positive species cannot dominate the reservoir phytoplankton solely because of specific phosphorus-scavenging strategy.

Triplophysa waisihani, a new species of nemacheiline loach from Northwest China (Pisces: Balitoridae)

Triplophysa waisihani, a new species of nemacheiline loach, is described from the Kax River, a tributary of the Ili River drainage in Xinjiang-Uighur Autonomous Region of Northwest China. It can be assigned to the T. labiata species group characterized by having widely separated anterior and posterior nostrils and no breeding tubercles on the sides of the head. Triplophysa waisihani resembles T. labiata and T. herzensteini, but differs from both in cephalic sensory-canal pattern and the structure of the gas bladder; from T. labiata in the structure of the pelvic girdle and absence or presence of the fourth basibranchial; and from T. herzensteini in the shape of the anal fin.

Macroinvertebrate community structure in Three-Gorges Reservoir, China

We investigated the macroinvertebrate community structure in Three-Gorges Reservoir during a three-year period after the reservoir became operational. Comparison with data of previous authors obtained before the damming showed that the benthic community changed drastically in the reservoir. Oligochaetes and chironomids dominate the present community. An apparent annual cycle in the benthic community (expressed as richness, density and community type) was found in the second year, lagging one year behind the annual cycle of transparency. After the second year, a Nais-Polypedilum community type, occurring in winter and spring with low inflow discharge and high transparency, was followed by a Limnodrilus community type that occurred in autumn and summer under contrasting conditions. Despite river regulation and damming, it appeared that macroinvertebrates in subtropical canyon-shaped reservoirs remained influenced by floods.

Relationships among the contents of total phenolics, soluble carbohydrate, and free amino acids of 15 aquatic macrophytes

In order to examine how carbon and nitrogen status of a macrophyte may affect its total phenolics (TP) production, the contents of free amino acids (FAA), soluble carbohydrate (SC) and TP were examined in leaves of seven submersed, four floating-leaved, and four emergent macrophytes. The floating-leaved and emergent macrophytes had much higher contents of SC and TP than the submersed macrophytes. The contents of FAA were not significantly different among the submersed, floating-leaved, and emergent macrophytes. Correlations among the contents of FAA, SC, and TP indicated that the production of TP was more dependent on the SC content than on the FAA content.

Phosphorus fractions and alkaline phosphatase activity in sediments of a large eutrophic Chinese lake (Lake Taihu)

Spatial, vertical, and seasonal variations in phosphorus fractions and in alkaline phosphatase activity (APA) were investigated in sediments in a large-shallow eutrophic Chinese lake (Lake Taihu) in 2003-2004. The phosphorus content was highest in the most seriously polluted lake area. Iron-bound phosphorus (Fe(OOH)-P) dominated (47% on average) among the phosphorus fractions determined according to Golterman (Hydrobiologia 335:87-95, 1996). Notably, organically-bound P comprised a further significant additional portion (acid-soluble + hot NaOH-extractable organic P = 25%), which was highest at the most polluted sites. The Fe(OOH)-P content was the lowest in spring (April, 2004), suggesting that degradation of organic matter led to the release of iron-bound phosphates. Sediment APA showed a significant positive relationship with both organically-bound P and Fe(OOH)-P. Consequently, organically-bound P is an important portion of the sediment phosphorus in Lake Taihu. It is mainly derived from freshly-settled autochthonous particles and from external discharges. Organically-bound P induces APA and may lead to the release of bioavailable phosphates from the organic sediments, thereby accelerating lake eutrophication.

Morphological variability between wild populations and inbred stocks of a Chinese minnow, Gobiocypris rarus

Gobiocypris rarus, a small, native cyprinid fish, is currently widely used in research on fish pathology, genetics, toxicology, embryology, and physiology in China. To develop this species as a model laboratory animal, inbred strains have been successfully created. In this study, to explore a method to discriminate inbred strains and evaluate inbreeding effects, morphological variation among three wild populations and three inbred stocks of G. rarus was investigated by the multivariate analysis of eight meristic and 30 morphometric characters. Tiny intraspecific variations in meristic characters were found, but these were not effective for population distinction. Stepwise discriminant analysis and cluster analysis of conventional measures and truss network data showed considerabe divergence among populations, especially between wild populations and inbred stocks. The average discriminant accuracy for all populations was 82.1% based on conventional measures and 86.4% based on truss data, whereas the discriminant accuracy for inbred strains was much higher. These results suggested that multivariate analyses of morphometric characters are an effective method for discriminating inbred strains of G. rarus. Morphological differences between wild populations and inbred strains appear to result from both genetic differences and environmental factors. Thirteen characters, extracted from stepwise discriminant analysis, played important roles in morphological differentiation. These characters were mainly measures related to body depth and head size.

Effects of ammonium on growth, nitrogen and carbohydrate metabolism of Potamogeton maackianus A. Benn

Growth, nitrogen and carbohydrate metabolism in relation to eutrophication were studied for a submerged plant Potamogeton maackianus, a species common in East Asian shallow lakes. The plants were grown in six NH4+-N concentrations (0.05, 0.50, 1.00, 3.50, 5.00 and 10.00 mg/L) for six days. NH4+-N levels in excess of 0.50 mg/L inhibited the plant growth. The relationships between external NH4+-N availability and total nitrogen (TN), protein-N, free amino acid-N (FAA-N) and NH4+-N in plant tissues, respectively, conformed to a logarithmic model suggesting that a feedback inhibition mechanism may exist for ammonium uptake. The response of starch to NH4+-N was fitted with a negative, logarithmic curve. Detailed analysis revealed that the influx NH4+-N had been efficiently incorporated into organic-N and eventually stored as protein at the expense of starch accumulation. These data suggest that this species may be able to tolerate high levels of ammonium when dissolved oxygen is sufficient.

Icefish (Salangidae) as an indicator of anthropogenic pollution in freshwater systems using nitrogen isotope analysis

We investigated differences in delta N-15 of seston and icefishes from seven freshwater ecosystems with different trophic states in China. An increase of seston delta N-15 values was accompanied by an increase of total nitrogen and phosphorus concentrations. Significantly positive correlations were observed between delta N-15 of icefishes and delta N-15 of seston, total nitrogen and phosphorus concentrations. This study demonstrated that icefishes could be preferred indicators of anthropogenic contamination in test systems because they integrated waste inputs over long time periods and reflected the movement of waste through the pelagic food chain.

The role of NH4+ toxicity in the decline of the submersed macrophyte Vallisneria natans in lakes of the Yangtze River basin, China

Experimental and field studies were conducted to evaluate the effects of NH4+ enrichment on growth and distribution of the submersed macrophyte, Vallisneria natans L, in lakes of the Yangtze River in China, based on the balance between free amino acids (FAA) and soluble carbohydrates (SC) in the plant tissue. Increase of NH4+ rather than NO3- concentrations in the water column caused FAA accumulation and SC depletion of the plant. The plant showed a unimodal pattern of biomass distribution along both FAA/SC ratios and external NH4+ concentrations, indicating that a moderate NH4-N concentration (< 0.3 mg L-1) benefited the plant, whereas the high NH4-N concentration (> 0.56 mg L-1) eliminated the plant completely. Therefore, 0.56 mg NH4-N mg L-1 in the water column was taken as the upper limit for V. natans in lakes of the Yangtze River basin. The mesocosm experiment showed that at a high external NH4-N (0.81 mg L-1), V. natans failed to propagate with a loss of half SC content (5 mg g(-1) DW) in the rhizomes, indicating that the consumption of carbohydrates for detoxification of excess NH4+ into non-toxic FAA significantly diminished carbohydrate supply to the rhizomes. This might consequently inhibit the vegetative reproduction of the plant, and also might be an important cause for the decline and disappearance of the plant with eutrophication. The present study for the first time reports substantial ecophysiological evidences for NH4+ stress to submersed macrophytes, and indicates that NH4+ toxicity arising from eutrophication probably plays a key role in the deterioration of submersed macrophytes like V. natans.

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.

Community structure of small fishes in a shallow macrophytic lake (Niushan Lake) along the middle reach of the Yangtze River, China

This study describes the current status of the small fish community in Niushan Lake in China, and examines the spatial and seasonal variations of the community in relation to key environmental factors. Based on macrophyte cover conditions, the lake was divided into three major habitat types: (1) Potamogeton maackianus habitat, (2) Potamogeton maackianus and Myriophyllum spicatum habitat, and (3) uncovered or less-covered habitat. Fish were sampled quantitatively in the three habitat types by block nets seasonally from September 2002 to August 2003. A total of 10 469 individuals from 27 fish species were caught, among which 20 species were considered as small fishes. Rhodeus ocellatus, Paracheilognathus imberbis, Pseudorasbora parva, Micropercops swinhonis and Cultrichthys erythropterus were recognized as dominant small fishes according to their abundance and occurrence. It was noted that (1) small fishes predominated the total number of fish species in the lake, which reflected to some degree the size diminution phenomenon of fish resources; (2) many small fishes had plant detritus as their food item, which was consistent with the abundance of macrophyte detritus in the lake and implied the importance of detritus in supporting small fish secondary production. Canonical correspondence analysis suggested that the spatial distributions of most small fishes were associated with complex macrophyte cover conditions. Macrophyte biomass was positively correlated with species richness, diversity index and the catch per unit of effort (CPUE) of the fish community. Water depth had no significant effects on species diversity and distribution of the small fishes. Correspondence analysis revealed a higher occurrence of the small fishes and higher abundance of individuals in summer and autumn. Seasonal length-frequency distributions of several species indicated that more larval and juvenile individuals appeared in spring and summer. This study provides some baseline information which will be essential to long-term monitoring of small fish communities in the Yangtze lakes.

Contributions of phosphatase and microbial activity to internal phosphorus loading and their relation to lake eutrophication

Phosphatase may accelerate the process of lake eutrophication through improving phosphorus bioavailability. This mechanism was studied in three Chinese eutrophic shallow lakes (Lake Taihu, Lake Longyang,and Lake Lianhua). Phosphatase activity was related to the concentration of soluble reactive phosphorus (SRP) and chlorophyll a. Stability of dissolved phosphatase in reverse micelles may be attributed to molecular size, conformation and active residues of the enzyme. At the site with Microcystis bloomed in Lake Taihu, dissolved phosphatase activity was higher and more stable in micelles, SRP concentrations were lower in interstitial water, the contents of different forms of phosphorus and the amounts of aerobic bacteria were lower while respiration efficiency was higher in sediments. Phosphobacteria, both inorganic and organic and other microorganisms were abundant in surface water but rare in sediments. Therefore, internal phosphorus may substantially flux into water column by enzymatic hydrolysis and anaerobic release, together with mobility of bacteria, thereby initiating the bloom. In short, biological mechanism may act in concert with physical and chemical factors to drive the internal phosphorus release and accelerate lake eutrophication.

Vertical variation in dissolved alkaline phosphatase activity in a shallow eutrophic lake determined in reverse micelles

We evaluated the feasibility of microencapsulating dissolved alkaline phosphatase of a water body into reverse micelle systems prepared by hexadecyltrimethylammonium bromide as a surfactant in cyclohexane and 1-butanol as co-surfactant. The dissolved alkaline phosphatase activity within the micelle was described, including its kinetic parameters and the effects of pH and temperature on catalytic activity in surface, overlying and interstitial water of Lake Donghu. We found the similarities on the behavior of dissolved alkaline phosphatase of surface and interstitial water in reverse micelles, which was distinctly different from its behavior in the overlying water. This difference likely reflected the different origins of the dissolved alkaline phosphatase in the vertical profile of the lake. This system provides a novel tool with which to study the diversity and ecological significance of extracellular enzymes in aquatic environments.

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.

The temporal and spatial distribution, composition and abundance of Protozoa in Chaohu Lake, China: Relationship with eutrophication

Systematic investigations into the temporal and spatial distribution, composition and abundance of protozoa in two regions with different trophic levels in Chaohu Lake, a large, shallow and highly eutrophic freshwater lake in China, were conducted during 2002-2003. A total of 114 species of protozoa, including phytomastigophorans, zoomastigophorans, amoebae and ciliates, were identified from 120 polyurethane foam unit (PFU) samples exposed at four stations and from various types of natural substrates. Of the 114 taxa, 36 core species were found on PFU substrates and 23 of these were found on natural ones. Protozoan abundance and chemical physical parameters at nine sampling stations, four in the western lake and five in the eastern part, indicate trophic gradient changes along the lake. Seasonal variations in the species composition of major groups at littoral PFU sampling stations illustrate the effect of a severe algal bloom on the protozoan community structure. Temporal and spatial distributions of individual abundance as functions of water temperature and trophic status were revealed. This study demonstrates again that the PFU artificial substrate method samples protozoan communities more effectively than routine natural substrate methods. (c) 2005 Elsevier GmbH. All rights reserved.