With 47% land coverage in 2016, agricultural land was one of the largest terrestrial biomes in Germany. About 70% of the agricultural land was cropped area with associated pesticide applications. Agricultural land also represents an essential habitat for amphibians. Therefore, exposure of amphibians to agrochemicals, such as fertilizers and pesticides, seems likely. Pesticides can be highly toxic for amphibians, even a fraction of the original application rate may result in high amphibian mortality.
To evaluate the potential risk of pesticide exposure for amphibians, the temporal coincidence of amphibian presence on agricultural land and pesticide applications (N = 331) was analyzed for the fire-bellied toad (Bombina bombina), moor frog (Rana arvalis), spadefoot toad (Pelobates fuscus) and crested newt (Triturus cristatus) during spring migration. In 2007 and 2008, up to 80% of the migrating amphibians temporally coincided with pesticide applications in the study area of Müncheberg, about 50 km east of Berlin. Pesticide interception by plants ranged between 50 to 90% in winter cereals and 80 to 90% in winter rape. The highest coincidence was observed for the spadefoot toad, where 86.6% of the reproducing population was affected by a single pesticide in winter rape during stem elongation with 80% pesticide interception by plants. Late migrating species, such as the fire-bellied toad and the spadefoot toad, overlapped more with pesticide applications than early migrating species, such as the moor frog, did. Under favorable circumstances, the majority of early migrants may not coincide with the pesticide applications of arable fields during spring migration.
To evaluate the potential effect of pesticide applications on populations of the common frog (Rana temporaria), a landscape genetic study was conducted in the vinicultural area of Southern Palatinate. Due to small sample sizes at breeding sites within viniculture, several DNA sampling methods were tested. Furthermore, the novel repeated randomized selection of genotypes approach was developed to utilize genetic data from siblings for more reliable estimates of genetic parameters. Genetic analyses highlighted three of the breeding site populations located in viniculture as isolated from the meta-population. Genetic differentiation among breeding site populations in the viniculture (median pairwise FST=0.0215 at 2.34 km to 0.0987 at 2.39 km distance) was higher compared to genetic differentiation among breeding site populations in the Palatinate Forest (median pairwise FST=0.0041 at 5.39 km to 0.0159 at 9.40 km distance).
The presented studies add valuable information about the risk of pesticide exposure for amphibians in the terrestrial life stage and possible effects of agricultural land on amphibian meta-populations. To conserve endemic amphibian species and their (genetic) diversity in the long run, the risk assessment of pesticides and applied agricultural management measures need to be adjusted to protect amphibians adequately. In addition, other conservation measures such as the creation of new suitable breeding site should be considered to improve connectivity between breeding site populations and ensure the persistence of amphibians in the agricultural land.
Systemic neonicotinoids are one of the most widely used insecticide classes worldwide. In addition to their use in agriculture, they are increasingly applied on forest trees as a protective measure against insect pests. However, senescent leaves containing neonicotinoids might, inter alia during autumn leaf fall, enter nearby streams. There, the hydrophilic neonicotinoids may be remobilized from leaves to water resulting in waterborne exposure of aquatic non-target organisms. Despite the insensitivity of the standard test species Daphnia magna (Crustacea, Cladocera) toward neonicotinoids, a potential risk for aquatic organisms is evident as many other aquatic invertebrates (in particular insects and amphipods) display adverse effects when exposed to neonicotinoids in the ng/L- to low µg/L-range. In addition to waterborne exposure, in particular leaf-shredding invertebrates (= shredders) might be adversely affected by the introduction of neonicotinoid-contaminated leaves into the aquatic environment since they heavily rely on leaf litter as food source. However, dietary neonicotinoid exposure of aquatic shredders has hardly received any attention from researchers and is not considered during aquatic environmental risk assessment. The primary aim of this thesis is, therefore, (1) to characterize foliar neonicotinoid residues and exposure pathways relevant for aquatic shredders, (2) to investigate ecotoxicological effects of waterborne and dietary exposure on two model shredders, namely Gammarus fossarum (Crustacea, Amphipoda) and Chaetopteryx villosa (Insecta, Trichoptera), and (3) to identify biotic and abiotic factors potentially modulating exposure under field conditions.
During the course of this thesis, ecotoxicologically relevant foliar residues of the neonicotinoids imidacloprid, thiacloprid and acetamiprid were quantified in black alder trees treated at field relevant levels. A worst-case model – developed to simulate imidacloprid water concentrations resulting from an input of contaminated leaves into a stream – predicted only low aqueous imidacloprid concentrations (i.e., ng/L-range). However, the model identified dietary uptake as an additional exposure pathway relevant for shredders up to a few days after the leaves’ introduction into the stream. When test organisms were simultaneously exposed (= combined exposure) to neonicotinoids leaching from leaves into the water and via the consumption of contaminated leaves, adverse effects exceeded those observed under waterborne exposure alone. When exposure pathways were separated using a flow-through system, dietary exposure towards thiacloprid-contaminated leaves caused similar sublethal adverse effects in G. fossarum as observed under waterborne exposure. Moreover, the effect sizes observed under combined exposure were largely predictable using the reference model “independent action”, which assumes different molecular target sites to be affected. Dietary toxicity for shredders might, however, be reduced under field conditions since UV-induced photodegradation and leaching decreased imidacloprid residues in leaves and thereby the toxicity for G. fossarum. In contrast, both shredders were found unable to actively avoid dietary exposure. This thesis thus recommends considering dietary exposure towards systemic insecticides, such as neonicotinoids, already during their registration to safeguard aquatic shredders, associated ecosystem functions (e.g., leaf litter breakdown) and ultimately ecosystem integrity.