Xiuyun Cao

Shifts between ammonia-oxidizing bacteria and archaea in relation to nitrification potential across trophic gradients in two large Chinese lakes (Lake Taihu and Lake Chaohu)

Ammonia oxidation plays a pivotal role in the cycling and removal of nitrogen in aquatic ecosystems. Recent findings have expanded the known ammonia-oxidizing prokaryotes from Bacteria to Archaea. However, the relative importance of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) in nitrification is still debated. Here we showed that, in two large eutrophic lakes in China (Lake Taihu and Lake Chaohu), the abundance of AOA and AOB varied in opposite patterns according to the trophic state, although both AOA and AOB were abundant. In detail, from mesotrophic to eutrophic sites, the AOA abundance decreased, while the AOB increased in abundance and outnumbered the AOA at hypertrophic sites. In parallel, the nitrification rate increased along these trophic gradients and was significantly correlated with both the AOB abundance and the numerical ratio of AOB to AOA. Phylogenetic analysis of bacterial amoA sequences showed that Nitrosomonas oligotropha- and Nitrosospira-affiliated AOB dominated in both lakes, while Nitrosomonas communis-related AOB were only detected at the eutrophic sites. The diversity of AOB increased from mesotrophic to eutrophic sites and was positively correlated with the nitrification rate. Overall, this study enhances our understanding of the ecology of ammonia-oxidizing prokaryotes by elucidating conditions that AOB may numerically predominated over AOA, and indicated that AOA may play a less important role than AOB in the nitrification process of eutrophic lakes.

Shifting nutrient-mediated interactions between algae and bacteria in a microcosm: Evidence from alkaline phosphatase assay

The impacts of different nutrient additions (N + P, N + P + C, 4N + P, 4N + P + C, N + 2P) on the growth of algae and bacteria were studied in a microcosm experiment. Since alkaline phosphatase activity (APA) provides an indication of phosphorus deficiency, the higher value for algal APA in the treatments with excess nitrogen and for bacterial APA in the treatments with excess carbon suggested that, algal and bacterial phosphorus-limited status were induced by abundant nitrogen and carbon input, respectively. Bacterial phosphorus-limited status was weakened due to higher bacterial competition for phosphorus, compared to algae. In comparison with the bacterial and specific bacterial APA, higher values of algal and specific algal APA were found, which showed a gradual increase that coincided with the increase of chlorophyll a concentration. This fact indicated not only a stronger phosphorus demand by algae than by bacteria, but also a complementary relationship for phosphorus demand between algae and bacteria. However, this commensalism could be interfered by glucose input resulting in the decline of chlorophyll a concentration. Furthermore, the correlation between bacterial numbers and chlorophyll a concentration was positive in treatments without carbon and blurry in treatments with carbon. These observations validate a hypothesis that carbon addition can stimulate bacterial growth justifying bacterial nutrient demand, which decreases the availability of nutrients to algae and affects nutrient relationship between algae and bacteria. However, this interference would terminate after algal and bacterial adaption to carbon input.

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 lake 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 ∼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.

Predominance of ammonia-oxidizing archaea and nirK-gene-bearing denitrifiers among ammonia-oxidizing and denitrifying populations in sediments of a large urban eutrophic lake (Lake Donghu)

The coupled nitrification-denitrification process plays a pivotal role in cycling and removal of nitrogen in aquatic ecosystems. In the present study, the communities of ammonia oxidizers and denitrifiers in the sediments of 2 basins (Guozhenghu Basin and Tuanhu Basin) of a large urban eutrophic lake (Lake Donghu) were determined using the ammonia monooxygenase subunit A (amoA) gene and the nitrite reductase gene. At all sites of this study, the archaeal amoA gene predominated over the bacterial amoA gene, whereas the functional gene for denitrification nirK gene far outnumbered the nirS gene. Spatially, compared with the Tuanhu Basin, the Guozhenghu Basin showed a significantly greater abundance of the archaeal amoA gene but less abundance of the nirK and nirS genes, while there was no significant difference of bacterial amoA gene copy numbers between the 2 basins. Unlike the archaeal amoA gene, the nirK gene showed a significant difference in community structure between the 2 basins. Archaeal amoA diversity was limited to the water-sediment cluster of Crenarchaeota, in sharp contrast with nirK for which 22 distinct operational taxonomic units were found. Accumulation of organic substances were found to be positively related to nirK and nirS gene copy numbers but negatively related to archaeal amoA gene copy numbers, whereas the abundance of the bacterial amoA gene was related to ammonia concentration.

Dredging effects on P status and phytoplankton density and composition during winter and spring in Lake Taihu, China

Phytoplankton density and composition, together with phosphorus (P) concentrations and size-fractionated alkaline phosphatase activity (APA), were investigated in dredged and undredged zones in Lake Taihu from January to April 2004. P concentrations were also determined in the corresponding interstitial water. Enzyme Labeled Fluorescence (ELF) was used for localizing extracellular phosphatase on phytoplankton cell membranes in April. The increase in phytoplankton density was paralleled by a significant increase in soluble reactive P (SRP) concentrations in the water column and interstitial water at all sites from January to April, with chlorophyte gradually becoming dominant. In February, at the undredged site, more algae dominated by chlorophyte occurred in overlying water, rather than in the surface, coinciding with higher SRP concentrations in overlying and interstitial water. Therefore, P status in the bottom is important to phytoplankton development in terms of density and composition. Undredged sites had higher SRP concentrations in interstitial water than dredged sites. Furthermore, Higher APA was observed, accompanied by higher dissolved organic P (DOP) and lower total P at the undredged site in February. Enzymatic hydrolysis of DOP may have been an additional source of P for phytoplankton. In April, Schroederia sp. was ELF labeled in surface water at the dredged site, which showed markedly lower SRP concentration, but not at the undredged site with higher SRP concentration. Thus, the dredging might regulate algal density and composition in water column by reducing P bioavailability.

The effect of organic matter accumulation on phosphorus release in sediment of Chinese shallow lakes.

The effects of organic matter in sediment on phosphorus release were studied by field investigations in eight Chinese shallow freshwater lakes with different trophic status and a laboratory experiment. The sediment organic matter content paralleled the trophic status, ranging from 6.1 to 173.0 g kg(-1) (dry weight), with the mean value of 63.1 g kg(-1) (dry weight). It was positively proportional to Soluble reactive phosphorus concentration in the interstitial water in a form of exponential function, but inversely related to the sediment Fe/P ratio. The sediment alkaline phosphatase activity was significantly related not only to the organic matter content (r = 0.829, P < 0.01, n = 120), but also to the soluble reactive phosphorus concentration in interstitial water (r = 0.454, P < 0.01, n = 42). In the laboratory experiment, the addition of organic matter (dry materials of an aquatic macrophyte) into the sediment significantly enhanced alkaline phosphatase activity and soluble reactive phosphorus release. However, in the treatment with organic matter added and aeration, this release was generally prevented in spite of an increase in APA. Hence, sediment organic matter can effectively accelerate phosphorus release by enzymatic hydrolysis and anaerobic desorption. The latter mechanism seems to be more important.

The phosphorus release pathways and their mechanisms driven by organic carbon and nitrogen in sediments of eutrophic shallow lakes

To reveal phosphorus (P) release pathways from sediment and their mechanisms induced by organic matter enrichment, 116 sampling sites (including surface water and sediment) in 29 shallow lakes with different eutrophic degrees in Wuhan city, China, were investigated from July 2011 to November 2011. Empirical relationship and structural equation model indicated that the decomposition of total organic matter (TOM), including proteins (PRT), carbo-hydrates (CHO) and lipids (especially PRT) mediated by extracellular enzymes, accelerated the formation of anaerobic status. On the other hand, coupled nitrification-denitrification caused by ammonium (NH4+-N) accumulation due to PRT decomposition further aggravated anaerobic status and nitrate removal in terms of the increase of dehydrogenase activity (DHA). As a consequence, ferric iron was reduced to ferrous iron and soluble reactive phosphorus (SRP) was released from iron-bound phosphorus (Fe(OOH)~P) in sediments. In addition, extracellular alkaline phosphatase can be induced by organic carbon and nitrogen on condition that the input of nitrogen (N) and carbon (C) exceeded by far that of P. Taken together, enrichment of N and C can result in P release through the formation of anaerobic status and alkaline phosphatase production. Hence, we indicated that a close coupling existed among C, N and P cycles.

Isolation and Characterization of Organic Phosphorus-Mineralizing Bacteria in Sediment of a Chinese Large Shallow Eutrophic Lake (Lake Taihu)

The organic phosphorus mineralizing bacteria (OPB) play an important role in phosphorus cycling in lake sediment, to which less attention has been paid. Monthly sediment samples in 2009 ending in October, together with the samples from different seasons (May, July, October, and December) in 2008, were collected at 6 sites in a Chinese large shallow eutrophic lake (Lake Taihu). The sediment OPB numbers ranged from 2.2 × 106 to 1.79 × 108 cells g−1 (dry weight) at different sampling sites and in different seasons, with the average being 3.88 × 107 cells g−1 (dry weight). Its number was highest at the most polluted site and peaked in spring and summer, which can be explained by the enrichment of organic matter in sediment. Furthermore, there existed a significant positive relationship between the OPB numbers and alkaline phosphatase activities in the sediment. The 6 OPB strains isolated from the sediment were distinct in terms of their colony morphology on the yolk agar, biochemical characteristics and phosphorus release abilities. According to the 16S rDNA sequences, these OPB belong to Bacillus cereus, Stenotrophomonas maltophilia, Stenotrophomonas sp., Bacillus cereus, Xanthomonas sp., Pseudomonas sp. They were distinguished from the OPB species recorded in a Chinese small shallow eutrophic lake whose sediment organic content was remarkably higher. Taken together, organic matter shaped the OPB community not only quantitatively but also qualitatively, which in concert facilitated the enzymatic hydrolysis of organic phosphorus in lake sediment.

Variations in kinetics of alkaline phosphatase in sediments of eutrophic, shallow, Chinese lakes

Kinetics of alkaline phosphatase in sediments of a shallow Chinese freshwater lake (Lake Donghu) were investigated. Spatially, among 20 sites sampled, Vmax and Km values of alkaline phosphatase in surface sediments were higher in the zone adjacent to sites with the highest chlorophyll a concentrations. Vertically, there was a peak in Vmax at intermediate sediment depths in addition to the expected maximum at the surface. Some inhibitors, such as CuSO4, ZnSO4 and Na2WO4, showed significantly different effects on kinetics of alkaline phosphatase in interstitial water and sediments. Moreover, alkaline phosphatase in interstitial water and sediments responded to Na2WO4 in different ways in Lake Taihu. These observations imply that the enzyme is immobilized in sediments, which became more stable with accelerated eutrophication, as suggested by highest alkaline phosphatase activity (APA) in sediments corresponding with highest water column chlorophyll a concentrations in Lake Donghu.

Seasonal Variations in Chlorophyll a Concentrations in Relation to Potentials of Sediment Phosphate Release by Different Mechanisms in a Large Chinese Shallow Eutrophic Lake (Lake Taihu)

The relationship between chlorophyll a and fractionation of sediment phosphorus, inorganic phosphate-solubilizing bacteria (IPB), and organic phosphate-mineralizing bacteria (OPB) was evaluated in a large Chinese shallow eutrophic lake (Lake Taihu) and its embayment (Wuli Bay). At the three study sites, the increase of chlorophyll a concentrations in April paralleled those of the iron bound phosphate accounting for major portion of sediment inorganic phosphate, and in June significantly higher OPB and IPB numbers (especially OPB) in sediment were main contributors to the peaks of chlorophyll a concentration. Even though IPB peaked from February to June, it should serve as an unimportant P source due to the irrelevancy with chlorophyll a and soluble reactive phosphorus (SRP). By contrast, at the other site in the embayment, the calcium-bound phosphate was predominant and solid, which was difficult to be released, and neither IPB nor OPB were detectable in the sediment, indicating weak potential for phosphorus release from the sediment, which was reflected in the small seasonal variation in SRP concentration in water column. Hence, the extents to which the three general mechanisms behind phosphate release from sediment (desorption of iron bound phosphate, solubilization by IPB and enzymatic hydrolysis by OPB) operated were different depending on seasons and sites in Lake Taihu, they may jointly drive phosphate release and accelerate the eutrophication processes.