Many classical therapeutants are going to be banned in Europe, and an urgent need for alternatives is emerging. This issue can be exemplified by one major parasitic disease in aquaculture and ornamental fish breeding: velvet disease. This disease, caused by dinoflagellates of the genera Amyloodinium and Piscinoodinium, is an important infection affecting cultured freshwater and marine ornamental and food fish, and consistently causes great financial loss to the associated industries. Therapeutants available contain copper, malachite green, or methylene blue, and which can be toxic to non‐target organisms in the surrounding environment. As a result, these chemicals are banned for use by the aquaculture industry in several countries, and a prohibition for commercial ornamental fishkeeping is likely to follow in most countries. Increasing development of resistance to therapeutants, and growing public awareness for animal welfare and environmental protection, have prompted research in the areas of alternative treatment options and immunostimulants. Hydrogen peroxide and peracetic acid are possible ‘green’ therapeutants which do not contribute residues to the environment. Natural feed supplements such as pre‐ and probiotics can increase animal welfare and prevent stress and/or infections. Humic substances are another promising, natural immunostimulants which will be considered in depth. The aim of this review is to provide an overview of risks and benefits of current treatment options and new approaches to replace harmful therapeutants and minimize the number of toxic residues discharged into the environment. Treatments will be discussed on various parasitic infections and focus, where available, on Amyloodinium and Piscinoodinium.

In recent years, biochars have gained increasing interest in mitigating climate changes and revitalizing contaminated or drained soil. Studies determining their impact on the ecosystem, especially on soil invertebrates, however, are still scarce and the neurotoxic potential of biochars has never been evaluated before. Using the model organism Caenorhabditis elegans we determined the neurotoxic effect of biochar produced from rice straw by pyrolysis at 500 °C at concentrations ranging from 0 to 2000 mg C∙L-1. Biochar had a hormetic effect on locomotion behavior. Furthermore, high concentrations impaired defecation as well as the recognition and response to a chemical attractant. None of the potential toxic chemicals in the biochar had sufficient high concentrations to explain the detected neurotoxic effect. Using electron paramagnetic resonance (EPR) spectroscopy, we detected free radicals in the biochar. Detrimental reaction of free radicals with biotic macromolecules can induce oxidative stress responses and are a potential reason for the evaluated neurotoxic effect of biochar. Overall, we were able to prove that biochars have the potential to act as weak neurotoxins to soil organisms and effects of persistent free radicals should be investigated further.

Aquaculture has become imperative to cover the demands for dietary animal protein. Simultaneously, it has to overcome prejudices from excessive use of antibiotics and environmental impacts. Natural supplements are traditionally applied orally. In this study, we demonstrated another pathway: the gills. Humic substances are immunostimulants and a natural part of every aquatic ecosystem, making them ideal to be used as bath stimulants. Five and 50 mg C/L of a fulvic acid-rich humic substance was added for 28 days to the water of juvenile rainbow trout (Oncorhynchus mykiss). This fulvic acid is characterized by a high content of phenolic moieties with persistent free radicals and a high electron exchange capacity. The high concentration of the fulvic acid significantly increased growth and reduced the food conversion ratio and the response to a handling-stressor. Phagocytosis and potential killing activity of head kidney leukocytes were increased, as well as the total oxyradical scavenging capacity (TOSC) and lysozyme activity in the gills. In conclusion, immunostimulation via gills is possible with our fulvic acid, and the high phenolic content improved overall health and stress resistance of fish.

Inflammation is an essential process as a reaction towards infections or wounding. Exposure to hazardous environmental pollutants can lead to chronic inflammations, where the resolving phase is delayed or blocked. Very contradictory studies have been reported on the pro- and anti-inflammatory effects of humic substances (HSs) leading to significant disagreements between researchers. To a certain extent, this can be attributed to the chemical heterogeneity of this group of xenobiotics. Here we show for the first time that pro- and anti-inflammatory effects can occur by one HSs. We adapted an assay that uses green fluorescence-labeled zebrafish larvae and CuSO₄ to indue an inflammation. In wild-type larvae, exposure to 50 µM CuSO₄ for 2 h activated the production of reactive oxygen species, which can be monitored with a fluorescence dye (H2DCFDA) and a microplate reader. This allows not only the use of wild-type fish but also a temporal separation of copper exposure and inflammatory substance while retaining the high throughput. This modified assay was then used to evaluate the inflammatory properties of a fulvic acid (FA). We found, that the aromatic structure of the FA protects from inflammation at 5 and 50 mg C/L, while the persistent free radicals enhance the copper-induced inflammation at ≥ 300 mg C/L.