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Subterranean ecosystems are among the most widespread environments on Earth, yet we still have poor knowledge of their biodiversity. To raise awareness of subterranean ecosystems, the essential services they provide, and their unique conservation challenges, 2021 and 2022 were designated International Years of Caves and Karst. As these ecosystems have traditionally been overlooked in global conservation agendas and multilateral agreements, a quantitative assessment of solution-based approaches to safeguard subterranean biota and associated habitats is timely. This assessment allows researchers and practitioners to understand the progress made and research needs in subterranean ecology and management. We conducted a systematic review of peer-reviewed and grey literature focused on subterranean ecosystems globally (terrestrial, freshwater, and saltwater systems), to quantify the available evidence-base for the effectiveness of conservation interventions. We selected 708 publications from the years 1964 to 2021 that discussed, recommended, or implemented 1,954 conservation interventions in subterranean ecosystems. We noted a steep increase in the number of studies from the 2000s while, surprisingly, the proportion of studies quantifying the impact of conservation interventions has steadily and significantly decreased in recent years. The effectiveness of 31% of conservation interventions has been tested statistically. We further highlight that 64% of the reported research occurred in the Palearctic and Nearctic biogeographic regions. Assessments of the effectiveness of conservation interventions were heavily biased towards indirect measures (monitoring and risk assessment), a limited sample of organisms (mostly arthropods and bats), and more accessible systems (terrestrial caves). Our results indicate that most conservation science in the field of subterranean biology does not apply a rigorous quantitative approach, resulting in sparse evidence for the effectiveness of interventions. This raises the important question of how to make conservation efforts more feasible to implement, cost-effective, and long-lasting. Although there is no single remedy, we propose a suite of potential solutions to focus our efforts better towards increasing statistical testing and stress the importance of standardising study reporting to facilitate meta-analytical exercises. We also provide a database summarising the available literature, which will help to build quantitative knowledge about interventions likely to yield the greatest impacts depending upon the subterranean species and habitats of interest. We view this as a starting point to shift away from the widespread tendency of recommending conservation interventions based on anecdotal and expert-based information rather than scientific evidence, without quantitatively testing their effectiveness., (© 2022 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.)

Groundwater fauna (stygofauna) comprises organisms that have adapted to the dark subterranean environment over a course of thousands and millions of years, typically having slow metabolisms and long life cycles. They are crucial players in the groundwater of oxygenic aquifers, and contribute to various ecosystem services. Today's knowledge of their sensitivity to anthropogenic impacts is incomplete and a critical analysis of the general relevance of local findings is lacking. In this review, we focus on those areas with the highest interference between humans and stygofauna: cities. Here is where local pollution by various contaminants and heat strongly stresses the unique groundwater ecosystems. It is demonstrated that it is difficult to discern the influence of individual factors from the findings reported in field studies, and to extrapolate laboratory results to field conditions. The effects of temperature increase and chemical pollution vary strongly between tested species and test conditions. In general, previous findings indicate that heating, especially in the long-term, will increase mortality, and less adapted species are at risk of vanishing from their habitats. The same may be true for salinity caused by road de-icing in cold urban areas. Furthermore, high sensitivities were shown for ammonium, which will probably be even more pronounced with rising temperatures resulting in altered biodiversity patterns. Toxicity of heavy metals, for a variety of invertebrates, increases with time and chronic exposure. Our current knowledge reveals diverse potential impacts on groundwater fauna by urban pollution, but our insights gained so far can only be validated by standardized and long-term test concepts., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

We present field data on the effects of heavy rainfall after drought on the mobility of glyphosate and redox conditions in a clayey floodplain soil. By applying glyphosate together with deuterated water as conservative tracer in combination with time resolved in situ redox potential measurements, the spatial and temporal patterns of water infiltration and pesticide transport as well as the concomitant changes of the redox conditions were revealed. Our findings demonstrate that shrinkage cracks in dry soils can serve as effective transport paths for atmospheric oxygen, water and glyphosate. The rain intensity of a typical summer storm event (approx. 25 mm within one hour) was sufficient to translocate deuterated water and glyphosate to the subsoil (50 cm) within 2 hours. Soil wetting induced partial closure of the shrinkage cracks and stimulated microbial activity resulting in pronounced dynamics of in situ soil redox conditions. Redox potentials in 40 to 50 cm depth dropped permanently to strongly reducing conditions within hours to days but fluctuated between reducing and oxidizing conditions in 10 to 30 cm depth. Our findings highlight the close link between the presence of macropores (shrinkage cracks), heavy rainfall after drought, redox dynamics and pesticide translocation to the subsoil and thus call for further studies addressing the effects of dynamic redox conditions as a limiting factor for glyphosate degradation.

Viruses, including human pathogenic viruses, can persist in water. For producing drinking water from surface water via bank filtration, natural attenuation capacities and the fate of viruses during the passage of aquatic sediments are of particular interest. Moreover, the increasing frequency of extreme hydrological events necessitate re-evaluation of the sustainability and efficacy of processes removing viruses. For this purpose, we performed bank sediment filtration experiments using a mesocosm in a technical-scale experimental facility that simulates a field situation under more tightly controlled conditions. We used the bacteriophage MS2 as a surrogate for enteric viruses to study the transport of different viral loads through the bank sediment. Additionally, we simulated a heavy rain event to investigate the re-mobilization of initially attached virus particles. We quantified the abundance of infectious MS2 phages by plaque assay and the total number of MS2 particles by qPCR. Also, we differentiated pore water concentrations by depths of the sediment column and investigated attachment to the sediment matrix at the end of the individual experimental phases. Bank filtration over a vertical distance of 80 cm through sandy sediment revealed a virus removal efficiency of 0.8 log 10 for total MS2 particles and 1.7 log 10 for infectious MS2 particles, with an initial phage concentration of 1.84 × 10 8 gene copies mL -1 . A low load of infectious MS2 (1.9 × 10 6 plaque forming units mL -1 ) resulted in a greater removal efficiency (3.0 log 10 ). The proportion of infectious MS2 phages of the total MS2 particle mass steadily decreased over time, i.e., in the course of individual breakthrough curves and with sediment depth. The simulated pulse of rainwater caused a front of low ionic strength water which resulted in pronounced phage remobilization. The high proportion of infectious MS2 among the detached phages indicated that attachment to the sediment matrix may substantially conserve virus infectivity. Therefore, the re-mobilization of previously attached viruses owing to hydrological extremes should be considered in water quality assessment and monitoring schemes., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Riverbank filtration is an established and quantitatively important approach to mine high-quality raw water for drinking water production. Bacterial fecal indicators are routinely used to monitor hygienic raw water quality, however, their applicability in viral contamination has been questioned repeatedly. Additionally, there are concerns that the increasing frequency and intensity of meteorological and hydrological events, i.e., heavy precipitation and droughts leading to high and low river levels, may impair riverbank filtration performance. In this study, we explored the removal of adenovirus compared with several commonly used bacterial and viral water quality indicators during different river levels. In a seasonal study, water from the Rhine River, a series of groundwater monitoring wells, and a production well were regularly collected and analyzed for adenovirus, coliphages, E. coli, C. perfringens, coliform bacteria, the total number of prokaryotic cells (TCC), and the number of virus-like particles (TVPC) using molecular and cultivation-based assays. Additionally, basic physico-chemical parameters, including temperature, pH, dissolved organic carbon, and nutrients, were measured. The highest log 10 reduction during the >72 m of riverbank filtration from the river channel to the production well was observed for coliforms (>3.7 log 10 ), followed by E. coli (>3.4 log 10 ), somatic coliphages (>3.1 log 10 ), C. perfringens (>2.5 log 10 ), and F+ coliphages (>2.1 log 10 ) at high river levels. Adenovirus decreased by 1.6-3.1 log units in the first monitoring well (>32 m) and was not detected in further distant wells. The highest removal efficiency of adenovirus and most other viral and bacterial fecal indicators was achieved during high river levels, which were characterized by increased numbers of pathogens and indicators. During low river levels, coliforms and C. perfringens were occasionally present in raw water at the production well. Adenovirus, quantified via droplet digital PCR, correlated with E. coli, somatic coliphages, TCC, TVPC, pH, and DOC at high river levels. At low river levels, adenoviruses correlated with coliforms, TVPC, pH, and water travel time. We conclude that although standard fecal indicators are insufficient for assessing hygienic raw water quality, a combination of E. coli, coliforms and somatic coliphages can assess riverbank filtration performance in adenovirus removal. Furthermore, effects of extreme hydrological events should be studied on an event-to-event basis at high spatial and temporal resolutions. Finally, there is an urgent need for a lower limit of detection for pathogenic viruses in natural waters. Preconcentration of viral particles from larger water volumes (>100 L) constitutes a promising strategy., (Copyright © 2021. Published by Elsevier Ltd.)