The Yi River, the second longest river in Shandong Province, China, flows through Linyi City and is fed by three tributary rivers, Beng River, Liuqing River, and Su River in the northeastern part of the city. In this study, we determined the concentrations of five heavy metals (Cr, Ni, Cu, Zn, and Pb) in water, sediment, and aquatic macrophyte samples collected from the junction of the four rivers and evaluated the potential ecological risk of heavy metal pollution. Most of the heavy metals in water were in low concentrations with the water quality index (WQI) below 1, suggesting low metal pollution. The sediments showed low heavy metal concentrations, suggesting a low ecological risk based on the potential ecological risk index (RI) and the geo-accumulation index (I_geo). The aquatic plant species Potamogeton crispus accumulated considerable amounts of heavy metals, which were closely related to the metal concentrations of the sediment. The plant species Salvinia natans also showed an excellent metal accumulation capability. Based on our results, the junction of the four rivers is only slightly polluted in terms of heavy metals, and the plant species P. crispus is a suitable bioindicator for sediment heavy metal pollution.

In order to improve the performance of methane production from agro-waste, a batched four-stage–two-phase anaerobic digestion (4S2P-AD) system was designed to combine the advantages of both anaerobic co-digestion (co-AD) and two-phase AD. The initial separation of two phases was performed using rice straw (RS) as a feedstock in acidogenic phase and cow manure (CM) in methanogenic phase at low inoculum/substrate (I/S) ratios of 0.5 and 0.2 and a high organic loading of 60 g volatile solid (VS)/L. The periodic round-trip reflux of leachate during the 4S2P-AD process facilitated re-inoculation throughout the four stages. The results showed that this round-trip reflux also dispersed toxic ammonia, balanced the carbon/nitrogen ratio, unified the microbial community structure, and led to the selection of Methanosarcina (relative abundance > 80%) as the dominant methanogens. With the abilities to overcome volatile fatty acid accumulation, shorten lag times, improve biodegradability, and foster synergistic effects, it was verified that the 4S2P-AD process can maintain efficient and stable methanogenesis from high-solid lignocellulosic feedstock. The averaged methane production throughout the four stages of 4S2P-AD was 234 mL/g VS. This result is 96% higher than the averaged methane production obtained from the four one-step AD groups using mono-feedstock, and 91% higher than that obtained using co-feedstock. This study provides a scientific reference for the development of new processes of bio-methane production from agro-waste with a high fermentation capacity and stability in the future.