Comparison and optimization of extraction protocol for intracellular phosphorus and its polyphosphate in enhanced biological phosphorus removal (EBPR) sludge.

Intracellular polyphosphate (poly-P) plays important roles in Enhanced biological phosphorus removal (EBPR) process, but an effective and reliable protocol for extracting intracellular P and its poly-P in EBPR sludge without hydrolysis of poly-P has not been setup yet. In the study, it was revealed that the severe hydrolysis of intracellular poly-P occurred during the different extraction processes, such as acid (i.e., HClO4, H2SO4 and HCl), basic (i.e., NaOH and KOH) and freezing-grind (under different solid-liquid ratios), but it did not occur during ultrasonic extraction process. The optimal extraction process of the ultrasonic protocol was 10 w/mL of ultrasonic power density and 15 min of ultrasonic time, when the extraction efficiency of intracellular P was 88.24 ±â€¯1.56%. In addition, the extraction efficiency of intracellular P could be furtherly improved by that the 0.75 mol/L LiCl solution was used to resuspend the bacterial cell before ultrasonic extraction (i.e., LiCl-ultrasonic protocol). The ultrasonic protocol was more suitable to extract the intracellular P and its poly-P of EBPR sludge than the other 4 protocols (i.e., PCA-NaOH, EDTA-NaOH, freezing-grind and LiCl-ultrasonic), which had the technical characteristics of (i) with relatively high extraction efficiency of intracellular P, (ii) without hydrolysis of intracellular poly-P, (iii) with weak noise signal in 31P NMR spectrum and (iv) with simple extraction process and short extraction time. It was founded by the ultrasonic protocol that there was the high content (82.88%-89.79% of intracellular P content) of intracellular poly-P with long average chain length (376.4-383.2) in the EBPR sludges. Importantly, it was confirmed that the EBPR process was related to the combined action of extracellular and intracellular poly-P using a new fractionation method of P in EBPR sludge, which included the ultrasonic protocol at high power density for extracting the intracellular P and its poly-P.

Rapid changes in eastern Himalayan alpine flora with climate change.

PREMISE OF THE STUDY: With biodiversity and rates of climate change among the highest, the eastern Himalaya are critical for understanding the interaction of these two variables. However, there is a dearth of longitudinal data sets that address the effects of climate change on the exceptional alpine biodiversity of the Himalaya. METHODS: We established permanent alpine vegetation monitoring plots in three mountain chains of the Hengduan Mountains, the easternmost Himalaya, which have warmed 0.03-0.05°C yr-1 since 1985. Recently, we resampled plots (176 1-m2 quadrat plots and 88 sections of 11 summits in three Hengduan mountain chains) to measure changes in vegetation after 7 years. KEY RESULTS: Over 7 years, Tibetan alpine vegetation increased in number of species (+8 species/summit; +2.3 species/m2 ), in frequency (+47.8 plants/m2 ), and in diversity (+1.6 effective species/m2 ). Stepwise regressions indicated that warmer temperatures, southerly aspects, and higher elevations were associated with greater increases in these vegetation metrics. Unexpectedly, Himalayan endemic species increased (+1.4 species/m2 ; +8.5 plants/m2 ), especially on higher-elevation summits. In contrast, the increase in relative abundance of non-alpine species was greater at lower-elevation summits. Plants used by local Tibetans also increased (+1.3 species/m2 ; +32 plants/m2 ). CONCLUSIONS: As in other alpine areas, biodiversity is increasing with climate change in the Himalaya. Unlike other areas, endemic species are proliferating at the highest summits and are indicators of change.

The roles of loosely-bound and tightly-bound extracellular polymer substances in enhanced biological phosphorus removal.

Extracellular polymeric substances (EPS) have be founded to participate in the process of enhanced biological phosphorus removal (EBPR), but the exact role of EPS in EBPR process is unclear. In this work, the roles of loosely-bound EPS (LB-EPS), tightly-bound EPS (TB-EPS) and microbial cell in EBPR were explored, taking the activated sludge from 4 lab-scale A/O-SBR reactors with different temperatures and organic substrates as objects. It was founded that the P of EBPR activated sludge was mainly stored in TB-EPS, but the P of non-EBPR activated sludge was primarily located in microbial cell. The P release and uptake of EBPR activated sludge was attributed to the combined action of TB-EPS and microbial cell. Furthermore, TB-EPS played an more important role than microbial cell in EBPR process. With the analysis of 31P NMR spectroscopy, both polyP and orthoP were the main phosphorus species of TB-EPS in EBPR sludge, but only orthoP was the main phosphorus species of LB-EPS and microbial cell. During the anaerobic-aerobic cycle, the roles of LB-EPS, TB-EPS and microbial cell in transfer and transformation of P in EBPR sludge were obviously different. LB-EPS transported and retained orthoP, and microbial cell directly anaerobically released or aerobically absorbed orthoP. Importantly, TB-EPS not only transported and retained orthoP, but also participated in biological phosphorus accumulation. The EBPR performance of sludge was closely related with the polyp in TB-EPS, which might be synthesized and decomposed by extracellular enzyme.