[Dianchi Lake macroinvertebrate community succession trends and retrogressive analysis].

Historical records and data from yield surveys conducted in 2009 and 2010 were used to investigate macroinvertebrate community succession trends in Dianchi Lake. Species richness has declined from 57 in the 1980s to 32 in 2010, representing a species loss of 44%. Among the major benthic groups, the highest rate of loss was recorded for mollusks (75%) and aquatic insects (39%). Surveys in 2009 and 2010 across the lake revealed that the total density was 1776 ind/m2, comprising oligochaetes (1706 ind/m2) and chironomids (68 ind/m2). Over a nearly twenty-year span (1992-2010), the density and biomass of oligochaetes first increased sharply (1992-2002) and then declined gradually (2002-2010). Further, chironomids have decreased gradually while the proportion of abundant species has increased. Limnodrilus hoffmeisteri became the sole dominant species with an average relative abundance of 74.1%. Cosmopolitan species, such as Einfeldia sp., disappeared across the lake; instead, tolerant species such as Chironomus plumosus, Ch. attenuatus and Tanypus chinensis became the common. Mollusk community structure has become simpler and many native species have gone extinct. Species of concern include Margarya melanioides, M. mondi, M. mansugi and Cipangopaludina dianchiensis, all rated as critically endangered by the IUCN. We found that the Shannon-Wiener index declined in Dianchi Lake, particularly in Caohai Lake, from 2.70 in the 1950s to 0.30 in 2009 and 2010. Species richness and biodiversity was significantly negative correlated with total phosphorus and total nitrogen. Factors responsible for the benthic community retrogression described here include habitat destruction, lowering of water quality, outbreaks of blue-green algae, extinction of submerged plants and lack of germplasm resources.

[Anaerobic biodegradation of microcystin by bacterial community from sediment of Dianchi Lake].

Aerobic biodegradation has been identified as the main attenuation mechanism for microcystin, but the role of anaerobic microcystin biodegradation remains unclear. To elucidate this process, we assessed the potential for anaerobic microcystin LR biodegradation by sediment microbial community from Dianchi Lake and evaluated the effects of environmental factors and additional nutrient sources on the rates of anaerobic biodegradation. The results showed that microcystin LR was rapidly degraded from 5 mg/L to below detection limit within 2 days, demonstrating that the indigenous microorganisms can efficiently degrade microcystin LR under anaerobic conditions and can use microcystin LR as a sole nitrogen source. The rates of anaerobic microcystin LR biodegradation increased with increasing incubation temperature within the experimental range of 15-30 degrees C. Anaerobic microcystin LR biodegradation was slower (pH = 5.0) or even ceased (pH = 3.0) at acidic pH, but there was no difference in the rates at neutral (pH = 7.0) and alkaline (pH 9.0, 11.0) conditions. The addition of glucose decreased pH of the culture by producing acidic compounds and therefore significantly inhibited the anaerobic biodegradation of microcystin LR, but with the addition of NO3-, this inhibition disappeared. NO3- amendment also retarded the biodegradation of microcystin LR, demonstrating that NO3- was not used as a terminal electron acceptor. These findings suggest that anaerobic biodegradation might be another main attenuation mechanism for microcystin LR in sediments and present a significant bioremediation potential.

[Biodegradation of LAS in the anaerobic water of a representative inlet of Lake Dianchi].

Biodegradation and degradation kinetics of anion-surfactant (LAS) in the anaerobic water of a representative inlet (Haihe River) of Lake Dianchi under different incubation conditions were studied by the 'river die-away' test method. The influences of temperature, pH, initial concentration of LAS, aeration condition and added nutrients (NH4Cl or NaH2PO4) on the biodegradation of LAS in the water were investigated. The results demonstrate that LAS can be biodegraded by microorganisms in the water and that the percentage of degradation of LAS was more than 95% after 26 d. The biodegradation of LAS fit the second kinetic model. Incubation temperature, initial concentration of LAS, aeration and added nutrients (NH4Cl or NaH2PO4) can all affect the biodegradation of LAS. When the incubation temperature increased from 10 degrees C to 25 degrees C, the biodegradation rate (p) of LAS increased from 0.21 d(-1) to 0.90 d(-1). The LAS degradation rate increased from 0.72 d(-1) under anaerobic condition to 1.97 d(-1) under continuous aeration condition. The increased initial concentrations of LAS lead to decrease of the biodegradation rate. NaH2PO4 accelerated the degradation of LAS but added NH4Cl instead inhibited degradation. In our experiment, pH value (7.05-9.44) had little influence on the biodegradation of LAS.

[Subchronic toxic effects of fish muscle-bound MCLR].

Subchronic oral gavage toxicity of MCLR in water and in fish muscle was examined in male Balb/C mice for 13 weeks to assess the safety of aquatic products. The results showed that the liver coefficient (p < 0.05), the activities of ALT and AST (p < 0.01) increased significantly and distinct centrilobular to midzonal hepatucellular occurred after oral gavage of dissolved MCLR at a dose of 68.75 microg/kg (body weight), but neither influence on the activities of BUN and Cr nor histological changes on kidney were observed at any time point. In contrast, the administration of fish muscle-bound MCLR at the same dose resulted in no obvious subchronic toxicity in mice, except that the increase of liver coefficient (p < 0.05) and the activity of ALT (p < 0.01) can be observed only at the first week. It was concluded that the toxicity of fish muscle-bound MCLR was much lower than that of dissolved MCLR.

[Preparative purification and characterization of [Dha7] microcystin RR].

One variant of microcystins was isolated and purified with cyanobacteria natural bloom as the starting material, which was collected in Dianchi Lake, China. The separation protocol involved extraction of cyanobacterial cells by 75% aqueous methanol, isolation by reversed-phase flash chromatography, and purification by reversed-phase semipreparative HPLC. The structure and purity of purified microcystin was identified with electrospray ionization mass spectrometry, UV spectrophotometer, and analytical HPLC. The purified microcystin was assigned as [Dha7]MCRR (purity > 95%), which was a demethylated variant of MCRR. The structure of purified microcystin was identified as cyclo-(Ala-Arg-MeAsp-Arg-Adda-Glu-Dha) with molecular weight of 1023. There was a maximum absorbance at 239 nm in its UV spectrum (200-300 nm). This variant of microcystins occurred frequently, and sometimes could become the main variant in waterbloom from Dianchi Lake.

Selective control of toxic Microcystis water blooms using lysine and malonic acid: an enclosure experiment.

Three enclosures (10 x 10 x 1.5-1.3 m in depth) were set beside Dianch Lake, Kunming, People's Republic of China, for the period from July 28 to August 26, 2002. The enclosures were filled with cyanobacterial (Microcystis aeruginosa) water bloom-containing lake water. Lake sediment that contained macrophytes and water chestnut seeds was spread over the entire bottom of each enclosure. Initially, 10 g/m(2) of lysine was sprayed in Enclosure B, and 10 g/m(2) each of lysine and malonic acid were sprayed together in Enclosure C. Enclosure A remained untreated and was used as a control. The concentrations of lysine, malonic acid, chlorophyll a, and microcystin as well as the cell numbers of phytoplankton such as cyanobacteria, diatom, and euglena were monitored. On day 1 of the treatment, formation of cyanobacterial blooms almost ceased in Enclosures B and C, although Microcystis cells in the control still formed blooms. On day 7 Microcystis cells in Enclosure B that had been treated with lysine started growing again, whereas growth was not observed in Microcystis cells in Enclosure C, which had been treated with lysine and malonic acid. On day 28 the surface of Enclosure B was covered with water chestnut (Trapa spp.) and the Microcystis blooms again increased. In contrast, growth of macrophytes (Myriophllum spicatum and Potamogeton crispus) was observed in Enclosure C; however, no cyanobacterial blooms were observed. Lysine and malonic acid had completely decomposed. The microcystin concentration on day 28 decreased to 25% of the initial value, and the pH shifted from the initial value of 9.2 to 7.8. We concluded that combined treatment with lysine and malonic acid selectively controlled toxic Microcystis water blooms and induced the growth of macrophytes.

Regeneration of autologous tissue-engineered cartilage by using basic-fibroblast growth factor in vitro culture / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of the basic fibroblast growth factor (b-FGF) to regenerate an autologous tissue-engineered cartilage in vitro.</p><p><b>METHODS</b>The Cells were harvested from the elastic auricular cartilage of swine,and were plated at the concentration of 1 x 10(4) cells/cm2 , studied in vitro at two different media enviroments: Group I contained Ham's F-12 with supplements and b-FGF, Group II contained Ham's F-12 only with supplements. The passage 2 cells (after 12.75 +/- 1.26 days) were harvested and mixed with 30% pluronic F-127/Ham's F-12 at the concentration of 50 x 10(6) cells/ml. It was injected subcutaneously at 0.5 ml per implant. The implants were harvested 8 weeks after the vivo culture and examined with the histological stains.</p><p><b>RESULTS</b>The chondrocytes displayed morphologically similar to the fibroblasts in the media containing basic-FGF. The number of cell doublings (after 12.75 +/- 1.26 days) in vitro culture was as the following: Group I, 70; Group II, 5.4. Eight 8 weeks after the vivo autologous implantation, the average weight (g) and volume (cm3) in each group was as the following: Group I, 0.371 g/0.370 cm3 Group II, 0.179 g/0.173 cm3 (P < 0.01). With the b-FGF in vitro culture, the cells were expanded by 70 times after 2 weeks. Histologically, all of the engineered cartilage in the two groups were similar to the native elastic cartilage.</p><p><b>CONCLUSION</b>These results indicate that the basic-FGF could be used positively to enhance the quality and quantity of the seeding cells for the generation of the well-engineered cartilage.</p>