Environmental flow is vital for maintaining river ecosystem health and ensuring the normal growth of aquatic organisms. The wetted perimeter method is indeed very useful in the assessment of environmental flow due to consideration of stream forms and minimum flow for aquatic life habitat. In this study, a river with obvious seasonality and external water diversion was selected as the typical research object; taking Jingle, Lancun, Fenhe Reservoir, and Yitang hydrological sections as control sections, we improved the existing wetted perimeter method in three aspects: (1) We improved the selection of hydrological data series. The selected hydrological data series should be of a certain length and can well reflect the hydrological changes of wet, normal, and dry years. (2) Different from the traditional wetted perimeter method, which only gives one environmental flow value, the improved method calculates the environmental flow month by month. (3) The improved wetted perimeter method establishes the relationship between native fish survival and environmental flow. Results indicated that the improved wetted perimeter took the survival of the main fishes into consideration, the ratio of the calculated results by the slope method to the multi year average flow was greater than 10%, which can ensure the fishes\' habitat is not being destroyed, and the results are more reasonable. Furthermore, the monthly environmental flow processes obtained were better than the annual unified environmental flow value determined by the existing method and are consistent with the natural hydrological situation and water diversion situation of the river. This study shows that the improved wetted perimeter method is feasible for research of river environmental flow with strong seasonal and large variation of annual flow.

Classical methods, used for large scale treatments such as mechanical or chemical extractions, affect the integrity of extracted cytosolic protein by releasing proteases contained in vacuoles. Our previous experiments on flow processes electroextraction on yeasts proved that pulsed electric field technology allows preserving the integrity of released cytosolic proteins, by not affecting vacuole membranes. Furthermore, large cell culture volumes are easily treated by the flow technology. Based on this previous knowledge, we developed a new protocol in order to electro-extract total cytoplasmic proteins from microalgae (Nannochloropsis salina, Chlorella vulgaris and Haematococcus pluvialis). Given that induction of electropermeabilization is under the control of target cell size, as the mean diameter for N. salina is only 2.5 μm, we used repetitive 2 ms long pulses of alternating polarities with stronger field strengths than previously described for yeasts. The electric treatment was followed by a 24h incubation period in a salty buffer. The amount of total protein release was observed by a classical Bradford assay. A more accurate evaluation of protein release was obtained by SDS-PAGE. Similar results were obtained with C. vulgaris and H. pluvialis under milder electrical conditions as expected from their larger size.

Pulsed electric fields are used to induce membrane permeabilization on cells. In the case of species with cell wall (yeasts, microalgae), it was previously shown that when the pulse duration was several ms long, this resulted in a cytoplasmic soluble protein slow leakage. In this work, we show that a similar consequence can be obtained with different strains of E. coli. Experimental evidences of a resulting wall alteration are described. Pre-industrial flow process pilots are used. As the membrane electropermeabilization can be irreversible by applying a proper choice of the pulse parameters, this approach is used for bacterial inactivation in flow process. It is observed that sub-millisecond pulse trains are more cost effective than longer ones.