Anaerobic digestion (AD) is used to produce biogas and can offer a solution in waste management. Digestate, the AD by-product, can be applied to soil to improve fertility. However, the response of soil biological communities is not fully understood. There are mixed reports on its impact on earthworm survival. This study aimed to investigate digestate effects on earthworm mortality, and to elucidate potential mechanisms underlying it, if observed, after digestate application to soil. Juvenile and adult Allolobophora chlorotica were used as model organisms and added to microcosms prepared in a glasshouse trial. Five replicated treatments were: liquid Digestate; Osmotic-Stress (i.e. same salt concentration as digestate); Labile-C (i.e. same Biological Oxygen Demand as Digestate); Synthetic-Digestate a mixture of Osmotic-Stress and Labile-C (i.e. same salt concentration and BOD as digestate); and Water as the control. Treatments were applied at two different standardised rates equivalent to the digestate’s N content (i.e. 150 kg N ha⁻¹ eq. or 300 kg N ha⁻¹ eq.). The two development stages of A. chlorotica had different responses to treatments. Adult biomass was significantly greater in the Water control R150 treatment than in Digestate. Significantly lower juvenile biomass was observed in the Digestate R300 treatment than in the Labile-C and Water control treatments. The biomass of adults in the Labile-C R300 treatment was significantly greater than in the Digestate, Osmotic-Stress, Synthetic-Digestate and Water control treatments. Both life-stages exhibited a decline in biomass across all treatments, but the adults had higher mortality rates. The biomass of adults and juveniles declined, respectively, by 90% and 62% for Digestate applied at the lower rate, and by 96% and 90% at the higher rate. Whereas the abundance of adults and juveniles exhibited 80% and 24% drop at the lower rate, and a 90% and 84% drop at the higher rate. This study demonstrates that digestate can have negative impact on earthworm morbidity and mortality when applied to soil at 60% water filed pore space, with most of the total weight loss per pot due to reduced earthworm abundance. A likely hypothesis could be the osmotic stress induced by salts present in the digestate. However, there are other factors that interact with this effect, including possibly anaerobic impacts caused by high water content soils, as well as other mechanisms that have not been fully elucidated through this experimental design. Nevertheless, this work provides the basis for further ecotoxicology studies on the impact of digestate applied to soil. Further, while this works has shown that digestate can negatively impact A. chlorotica survival, whether the same is true for other earthworm species, ecotypes and life-cycle stages warrants further investigation. Considering the important role that worms play in soil health, field scale studies are also required to monitor the impacts of repeated digestate application on earthworm communities.

厌氧消化（AD）用于产生沼气，可以为废物管理提供解决方案。AD副产物Digestate可用于土壤以提高肥力。但是，对土壤生物群落的反应还没有完全了解。关于它对earth生存的影响的报道不一。这项研究旨在调查消化对on死亡率的影响，并阐明消化后施用到土壤后潜在的潜在机制。少年和成人Allolobophora chlorotica用作模型生物，并添加到在温室试验中制备的微观世界。五种重复处理是：液体消化剂；渗透胁迫（即盐浓度与消化液相同）；Labile-C（即与Digestate相同的生物需氧量）；合成消化法（Osmotic-Stress）和Labile-C的混合物（即与消化液相同的盐浓度和BOD）；以水为对照。以相当于消化液氮含量的两种不同的标准化速率（即150 kg N ha ^-1当量或300 kg N ha ^-1当量）进行处理。绿叶假单胞菌的两个发育阶段对治疗有不同的反应。在水控制R150处理中，成人生物量显着大于Digestate。在Digestate R300处理中观察到的幼生物量明显低于在Labile-C和水控制处理中。在Labile-C R300处理中，成年人的生物量显着大于Digestate，渗透压，合成Digestate和水控制处理。在所有治疗中，这两个生命阶段均表现出生物量下降，但成年人的死亡率更高。以较低的比例施用Digestate的成年人和青少年的生物量分别下降90％和62％，以较高的比例分别下降96％和90％。而成年人和少年的丰富度在较低的比率下下降了80％和24％，在较高的比率下下降了90％和84％。这项研究表明，如果将消化液施用到土壤含水量为60％的土壤中，消化液可能会对can的发病率和死亡率产生负面影响，而每盆的总重量损失大部分是由于most丰度降低所致。一个可能的假设可能是消化物中存在的盐引起的渗透压。但是，还有其他因素与此效应相互作用，包括高含水量土壤可能引起的厌氧影响，以及该实验设计尚未完全阐明的其他机理。尽管如此，这项工作为进一步研究消化物对土壤的影响提供了生态毒理学的基础。此外，尽管这项工作表明消化可能会对人体产生负面影响 由于pot含量降低，每个锅的总重量损失大部分。一个可能的假设可能是消化物中存在的盐引起的渗透压。但是，还有其他因素与此效应相互作用，包括高含水量土壤可能引起的厌氧影响，以及该实验设计尚未完全阐明的其他机理。尽管如此，这项工作为进一步研究消化物对土壤的影响提供了生态毒理学的基础。此外，尽管这项工作已经表明，消化会产生负面影响 由于pot含量降低，每个锅的总重量损失大部分。一个可能的假设可能是消化物中存在的盐引起的渗透压。但是，还有其他因素与此效应相互作用，包括高含水量土壤可能引起的厌氧影响，以及该实验设计尚未完全阐明的其他机理。尽管如此，这项工作为进一步研究消化物对土壤的影响提供了生态毒理学的基础。此外，尽管这项工作表明消化可能会对人体产生负面影响 包括高含水量土壤可能引起的厌氧影响，以及尚未通过此实验设计充分阐明的其他机理。尽管如此，这项工作为进一步研究消化物对土壤的影响提供了生态毒理学的基础。此外，尽管这项工作已经表明，消化会产生负面影响 包括高含水量土壤可能引起的厌氧影响，以及尚未通过此实验设计充分阐明的其他机制。尽管如此，这项工作为进一步研究消化物对土壤的影响提供了生态毒理学的基础。此外，尽管这项工作表明消化可能会对人体产生负面影响对于其他earth物种，生态型和生命周期阶段，A。chlorotica的存活情况是否同样如此，值得进一步研究。考虑到蠕虫在土壤健康中的重要作用，还需要进行田间规模研究，以监测重复施肥对earth群落的影响。