The potential use of biochar from olive mill waste for in situ remediation of metal contaminated soils was evaluated. Biochar was mixed with metal contaminated soil originating from the vicinity of an old zinc smelter. Soil-biochar mixtures were equilibrated for 30 and 90 days. At these time points, Ca(NO3)2 exchangeable metals were determined, and effects of the biochar amendment on soil toxicity were investigated using plants, bacteria, and earthworms. Bean (Phaseolus vulgaris) growth, metal content, antioxidative enzymes activities, and soluble protein contents were determined. Furthermore, effects on soil microbial communities (activity, diversity, richness) were examined using Biolog ECOplates. After 120 days of soil-biochar equilibration, effects on weight and reproduction of Eisenia foetida were evaluated. With increasing biochar application rate and equilibration period, Ca(NO3)2 exchangeable metals decreased, and growth of bean plants improved; leaf metal contents reduced, the activities of antioxidative stress enzymes decreased, and soluble protein contents increased. Soil microbial activity, richness, and diversity were augmented. Earthworm mortality lowered, and their growth and reproduction showed increasing trends.

No abstract

Biological invasions contribute now more than ever to the global homogenization of fauna and flora. Large‐scale monitoring programs are, therefore, needed to detect incipient invasions and to evaluate management interventions. As conventional monitoring methods are constrained by large costs, environmental DNA (eDNA)‐based methods are increasingly recognized as valuable monitoring tools. However, accurately estimating species abundance from eDNA concentrations in natural systems remains challenging and consequently hinders their integration in management applications. Here, we used droplet digital PCR (ddPCR) in eDNA surveys to estimate the abundance of invasive American bullfrogs (Lithobates catesbeianus). We first introduced bullfrog tadpoles in natural ponds to assess the relationship between abundances and eDNA concentrations under field conditions. Next, we combined eDNA sampling with fyke netting in naturally colonized ponds to investigate whether bullfrog eDNA concentrations can estimate bullfrog capture success and conventional abundance measures obtained via depletion sampling. Finally, we evaluated eradication measures by comparing bullfrog eDNA concentrations before and after fyke netting. We found a strong linear relationship between the numbers of introduced tadpoles and eDNA concentrations (r2 = 0.988). Bullfrog eDNA concentrations were not only linearly related to the catch‐per‐unit‐effort (r2 = 0.739), but also to conventional abundance estimates (r2 = 0.716), particularly when eDNA concentrations were standardized for pond area (r2 = 0.834) and volume (r2 = 0.888). Bullfrog tadpoles were only captured when eDNA concentrations exceeded 1.5 copies µl−1, indicating that quantitative eDNA analyses enable the localization of breeding ponds. We found a significant reduction in eDNA concentrations after fyke netting proportional to the number of captured bullfrogs. These results demonstrate that eDNA quantification is a reliable tool that accurately estimates bullfrog abundance in natural lentic systems. We show that quantitative eDNA analyses can complement the toolbox of natural resource managers and facilitate the coordination of eradication campaigns targeting alien invasive species.

Nine adult American bullfrogs (Lithobates catesbeianus) were tagged with an internal radio transmitter and tracked during one year in the valley of the Grote Nete (Belgium). The mean ± SD core range area (KDE50) was 15.00 ± 22.41m2. The home range area (KDE95) had a mean ± SD of 429.78 ± 510.97m2. Shores of larger eutrophic ponds and small temporary pools in alluvial forest were chosen as habitat. The total area used (MCP95) had a mean of 11,086.73 ± 12,239.00m2. The study revealed a mean action radius of 270.78 ± 199.17m and individuals moved up to 742m in a single displacement. These results show that the dispersion of the American bullfrog in a valley system such as the Grote Nete can proceed very rapidly. A positive correlation between weight and distance covered within one movement was found, which could suggest that dominant individuals are capable of covering greater distances in search of optimal habitat for reproduction, foraging or hibernation.