Dose-dependent in vivo effects of formulated moxidectin on seedling emergence of temperate grassland species.

Sheep function as effective endozoochorous seed vectors in grasslands. Recent laboratory-based studies showed that this important function can be impaired by macrocyclic lactone anthelmintics, which are used to control parasites and enter into the environment mainly via faeces; however, there is a lack of in vivo studies. We conducted a seed-feeding experiment with sheep that included four temperate grassland species from four different families (Achillea ptarmica, Asteraceae; Agrostis capillaris, Poaceae; Dianthus deltoides, Caryophyllaceae; Plantago lanceolata, Plantaginaceae). A series of three feeding trials was carried out after one of two groups of sheep received a single administration of a common oral formulation of the macrocyclic lactone moxidectin. Faeces were collected to determine seedling emergence rate and emergence timing as well as moxidectin concentration via HPLC. Seedling emergence differed significantly between the anthelmintic-treated sheep and the control group. This impact depended on time of seed uptake after anthelmintic administration. Number of emerging seedlings was significantly reduced (27.1 %) when faeces moxidectin concentrations were high (on average 3153 ng g-1; 1 d post treatment) and significantly increased (up to 68.8 %) when moxidectin concentrations were low (≤86 ng g-1; 7, 14 d pt). Mean emergence time was significantly lowered at low moxidectin concentrations. These results demonstrate dose-related effects of deworming on seedling emergence which might affect endozoochory and eventually plant population dynamics in grasslands.

Temperature moderates impact of formulated moxidectin on seed germination of three temperate grassland species.

Formulations of macrocyclic lactone anthelmintics such as moxidectin are regularly administered to sheep to combat parasites. A disadvantage of these pharmaceuticals are their side effects on non-target organisms when entering the environment. Little is known about anthelmintic effects on plant reproduction and whether the effects depend on environmental factors. For ecological and methodological reasons, we aimed at testing whether temperature affects the efficacy of a common moxidectin-based formulation on seed germination. We carried out a germination experiment including three typical species of temperate European grasslands (Centaurea jacea, Galium mollugo, Plantago lanceolata). We applied three temperature regimes (15/5, 20/10, 30/20°C), and a four-level dilution series (1:100-1:800) of formulated moxidectin (i.e., Cydectin oral drench). These solutions represent seed-anthelmintic contacts in the digestive tract of sheep shortly after deworming. In addition, a control was carried out with purified water only. We regularly counted emerging seedlings and calculated final germination percentage, mean germination time and synchrony of germination. Formulated moxidectin significantly reduced percentage, speed and synchrony of germination. A 1:100 dilution of the formulation reduced germination percentage by a quarter and increased mean germination time by six days compared to the control. Temperature moderated effects of the anthelmintic drug on germination in all response variables and all species, but in different patterns and magnitudes (significant anthelmintic x temperature x species interactions). In all response variables, the two more extreme temperature regimes (15/5, 30/20°C) led to the strongest effects of formulated moxidectin. With respect to germination percentage, G. mollugo was more sensitive to formulated moxidectin at the warmest temperature regime, whereas P. lanceolata showed the highest sensitivity at the coldest regime. This study shows that it is important to consider temperature dependencies of the effects of pharmaceuticals on seed germination when conducting standardised germination experiments.

The Anthelmintic Ingredient Moxidectin Negatively Affects Seed Germination of Three Temperate Grassland Species.

In animal farming, anthelmintics are regularly applied to control gastrointestinal nematodes. There is plenty of evidence that also non-target organisms, such as dung beetles, are negatively affected by residues of anthelmintics in faeces of domestic ungulates. By contrast, knowledge about possible effects on wild plants is scarce. To bridge this gap of knowledge, we tested for effects of the common anthelmintic formulation Cydectin and its active ingredient moxidectin on seed germination. We conducted a feeding experiment with sheep and germination experiments in a climate chamber. Three wide-spread plant species of temperate grasslands (Centaurea jacea, Galium verum, Plantago lanceolata) were studied. We found significant influences of both, Cydectin and moxidectin, on germination of the tested species. Across species, both formulation and active ingredient solely led to a decrease in germination percentage and synchrony of germination and an increase in mean germination time with the formulation showing a more pronounced response pattern. Our study shows for the first time that anthelmintics have the potential to negatively affect plant regeneration. This has practical implications for nature conservation since our results suggest that treatments of livestock with anthelmintics should be carefully timed to not impede endozoochorous seed exchange between plant populations.