Linking landscape composition and biological factors with exposure levels of rodenticides and agrochemicals in avian apex predators from Germany.

Intensification of agricultural practices has resulted in a substantial decline of Europe's farmland bird populations. Together with increasing urbanisation, chemical pollution arising from these land uses is a recognised threat to wildlife. Raptors are known to be particularly sensitive to pollutants that biomagnify and are thus frequently used sentinels for pollution in food webs. The current study focussed on anticoagulant rodenticides (ARs) but also considered selected medicinal products (MPs) and frequently used plant protection products (PPPs). We analysed livers of raptor species from agricultural and urban habitats in Germany, namely red kites (MIML; Milvus milvus), northern goshawks (ACGE; Accipiter gentilis) and Eurasian sparrowhawks (ACNI; Accipiter nisus) as well as white-tailed sea eagles (HAAL; Haliaeetus albicilla) and ospreys (PAHA; Pandion haliaetus) to account for potential aquatic exposures. Landscape composition was quantified using geographic information systems. The highest detection of ARs occurred in ACGE (81.3%; n = 48), closely followed by MIML (80.5%; n = 41), HAAL (38.3%; n = 60) and ACNI (13%; n = 23), whereas no ARs were found in PAHA (n = 13). Generalized linear models demonstrated (1) an increased probability for adults to be exposed to ARs with increasing urbanisation, and (2) that species-specific traits were responsible for the extent of exposure. For MPs, we found ibuprofen in 14.9% and fluoroquinolones in 2.3% in individuals that were found dead. Among 30 investigated PPPs, dimethoate (and its metabolite omethoate) and thiacloprid were detected in two MIML each. We assumed that the levels of dimethoate were a consequence of deliberate poisoning. AR and insecticide poisoning were considered to represent a threat to red kites and may ultimately contribute to reported decreased survival rates. Overall, our study suggests that urban raptors are at greatest risk for AR exposure and that exposures may not be limited to terrestrial food webs.

Binding of RDX to Cell Wall Components of Pinus sylvestris and Picea glauca and Three-Year Mineralisation Study of Tissue-Associated RDX Residues.

Contamination of soils with the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX, Research Department Explosive) as a result of military applications is a large-area problem globally. Since coniferous trees dominate the vegetation of large areas of military land in Central Europe, particularly in Germany, the long-term fate of (14)C-RDX in the conifers Scots pine and Dwarf Alberta spruce was studied. Acetic acid was the most effective solvent for the removal of extractable RDX residues from homogenates of RDX-laden tree material (85%, 80-90% and 64-80% for roots, wood and needles, respectively). On average, only a fifth of RDX-derived (14)C was bound in non-extractable residues (NER). Within the main cell wall compartments, lignin was the dominant binding site for NER (needles: 32-62%; roots: 38-42%). Hemicellulose (needles: 11-18%; roots: 6-11%) and cellulose (needles: 12-24%; roots: 1-2%) were less involved in binding and a considerable proportion of NER (needles: 15-24%; roots: 59-51%) was indigestible. After three-year incubation in rot chambers, mineralisation of tree-associated (14)C-RDX to (14)CO2 clearly dominated the mass balance in both tree species with 48-83%. 13-33% of (14)C-RDX-derived radioactivity remained in an unleachable form and the remobilisation by water leaching was negligible (< 2%).

Effects of an Anti-Fertility Product on Reproductive Structures of Common Vole Males and Residues of Compounds.

Some rodent species cause significant damage to agriculture and forestry, and some can transmit pathogens to humans and livestock. The common vole (Microtus arvalis) is widespread in Europe, and its population outbreaks have resulted in massive crop loss. Bait-based fertility control could contribute to rodent pest management. Bait containing 4-vinylcyclohexene diepoxide (VCD) and triptolide (TP), registered as ContraPest®, was delivered to male common voles for 14 or 28 consecutive days. The effects on reproductive structures and residues in the liver and testes were assessed. There was no effect on testis weight, sperm viability, sperm motility and oxidative stress in sperm cells. Results regarding the mitochondrial membrane potential of sperm, DNA fragmentation and progressively motile sperm cells were inconclusive. However, there was an increase in morphological sperm defects in voles treated for 14/28 days and fewer normal sperm cells in voles treated for 28 days. There were no TP residues in the testes, few and low TP residues and no VCD residues in liver tissues, making considerable secondary exposure to non-target species unlikely. Treatments with VCD + TP seemed to have minor effects on the reproductive organs of males. Further studies should evaluate the effect of VCD + TP on females and on the reproductive success of common voles and other pest rodent species.

Comparison of baiting strategies in common vole management.

BACKGROUND: Worldwide, pest rodents can cause extensive damage to agriculture, forestry, food storage, and infrastructure and pose a risk to public health and livestock due to the spread of zoonotic pathogens. In Europe, the most common pest rodent species is the common vole (Microtus arvalis). Management during periodic outbreaks largely relies on rodenticidal bait with zinc phosphide. Efficient baiting with rodenticides or possibly anti-fertility products in the future require baiting methods that allow a sufficient proportion of the population to consume an effective dose of bait. We used a bait with the quantitative marker ethyl-iophenoxic acid (Et-IPA) to evaluate baiting strategies in enclosure experiments. This wheat-based bait with Et-IPA was placed in bait boxes or directly into the tunnel system entrances in different seasons and common vole abundances. Voles were live-trapped, individually marked and blood samples were collected to relate Et-IPA blood residues to bait uptake. RESULTS: The percentage of animals consuming bait was not heavily affected by the baiting strategy but voles had higher Et-IPA blood residues if tunnel baiting was used in autumn and if bait boxes were used in winter. Non-reproductive as well as lighter animals tended to have higher Et-IPA blood residues than reproductive individuals, whereas sex had no effect. Population density had a negative effect on the probability of residues present as well as on Et-IPA blood concentration. CONCLUSION: The results of this study might help to improve baiting techniques to manage overabundant rodent pest species regardless of the compounds to be delivered. © 2024 Julius Kühn-Institut. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Exposure assessment of anticoagulant rodenticides in the liver of red foxes (Vulpes vulpes) in Slovenia.

The study deals with the environmental residues of anticoagulant rodenticides (ARs) in Slovenia to evaluate the toxicological risk of secondary poisoning of red foxes (Vulpes vulpes) as representatives of non-target wildlife, and in relation to the investigated use patterns of ARs and specific local parameters in Slovenia. From 2019 to 2022, 148 liver tissue samples of adult red foxes were collected from almost all state geographical regions. The samples were extracted with methanol/water (2:1, v/v), cleaned-up using a solid supported liquid-liquid extraction, and measured by liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) with reporting limits of 0.5 to 5.0 ng/g. Residues of at least one rodenticide were detected in 77.7 % of the samples. The second generation ARs of bromadiolone, brodifacoum and difenacoum were the most frequently found, appearing in 75.0, 51.4, and 18.9 % of the samples, respectively. Concentrations of pooled ARs ranged from 1.5 to 2866.5 ng/g with mean and median values of 601.4 and 350.2 ng/g, respectively. We determined bromadiolone and brodifacoum at concentrations of ≥800 ng/g in 10.8 and 10.1 % of the samples, and 1.4 and 0.7 % of the samples contained residues >2000 ng/g, respectively. These concentrations are much higher than those found in comparable studies in Europe and elsewhere in the world. Residues of ARs were detected in all monitored statistical regions of Slovenia, with higher concentrations in the eastern parts of the country. First generation ARs were found in only 9.5 % of samples, and residues were below 10 ng/g with one exception (coumatetralyl with 55 ng/g). The results of the study indicate a serious toxicological risk for red foxes in Slovenia as part of the Western Balkans, and will contribute to the growing body of knowledge about the protection of European ecosystems, as wildlife is not limited by national borders.

Ethyl-iophenoxic acid as a quantitative bait marker for small mammals.

Bait markers are indispensable for ecological research but in small mammals, most markers are invasive, expensive and do not enable quantitative analyses of consumption. Ethyl-iophenoxic acid (Et-IPA) is a non-toxic, quantitative bait marker, which has been used for studying bait uptake in several carnivores and ungulates. We developed a bait with Et-IPA, assessed its palatability to common voles (Microtus arvalis), and determined the dose-residue-relation for this important agricultural pest rodent species. Et-IPA concentrations of 40 to 1280 µg Et-IPA per g bait were applied to wheat using sunflower oil or polyethylene glycol 300 as potential carriers. In a laboratory study, common voles were offered the bait and blood samples were collected 1, 7, and 14 days after consumption. The samples were analyzed with LC-ESI-MS/MS for blood residues of Et-IPA. Sunflower-oil was the most suitable bait carrier. Et-IPA seemed to be palatable to common voles at all test concentrations. Dose-dependent residues could be detected in blood samples in a dose-dependent manner and up to 14 days after uptake enabling generation of a calibration curve of the dose-residue relationship. Et-IPA was present in common vole blood for at least 14 days, but there was dissipation by 33-37% depending on dose. Et-IPA meets many criteria for an "ideal" quantitative bait marker for use in future field studies on common voles and possibly other small mammal species.

Noninvasively Collected Fecal Samples as Indicators of Multiple Pesticide Exposure in Wild Birds.

Pesticide use poses a potential hazard to wild birds that use agricultural farmland as their foraging habitat. Whereas most current pesticide studies have found residues in liver samples and single active substances, noninvasive sampling methods and data on a wide variety of agrochemicals are needed to determine pesticide exposure of living wild birds for postregistration monitoring. We collected feces during autumn migration of Eurasian skylarks (Alauda arvensis), a species that commonly forages in winter cereal crops. Birds were kept in paper bags until we measured their body condition, individually marked and released them. We analyzed the feces dropped in paper bags for the presence of 80 pesticides including rodenticides and degradation products. Nine active substances from fungicides and herbicides commonly used in grain and maize fields were detected individually, or in combination, in 25% of the samples. We found no significant differences in body condition between exposed and unexposed birds, but Eurasian skylarks without pesticide residues had a better body condtion score on average than birds with pesticide residues. Pesticide determination in noninvasively collected fecal samples allows a refined risk analysis, which takes pesticides used in the habitats of birds into account. It allows the search for the sources of pesticide contamination, but also enables research into potential deleterious effects on the fitness of farmland birds. Environ Toxicol Chem 2022;41:201-207. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Spatial variation of rodenticides and emerging contaminants in blood of raptor nestlings from Germany.

Wildlife exposures to pest controlling substances have resulted in population declines of many predatory species during the past decades. Many pesticides were subsequently classified as persistent, bioaccumulative, and toxic (PBT) and banned on national or global scales. However, despite their risks for non-target vertebrate wildlife, PBT substances such as anticoagulant rodenticides (ARs) are still permitted for use in Europe and have shown to threaten raptors. Whereas risks of ARs are known, much less information is available on emerging agrochemicals such as currently used PPPs and medicinal products (MPs) in higher trophic level species. We expect that currently used PPPs are relatively mobile (vs. lipophilic) as a consequence of the PBT criteria and thus more likely to be present in aqueous matrices. We therefore analyzed blood of 204 raptor nestlings of three terrestrial (red kite, common buzzard, Montagu's harrier) and two aquatic species (white-tailed sea eagle, osprey) from Germany. In total, we detected ARs in 22.6% of the red kites and 8.6% of the buzzards, whereas no Montagu's harriers or aquatic species were exposed prior to sampling. ΣAR concentration tended to be higher in North Rhine-Westphalia (vs. North-Eastern Germany) where population density is higher and intense livestock farming more frequent. Among the 90 targeted and currently used PPPs, we detected six substances from which bromoxynil (14.2%) was most frequent. Especially Montagu's harrier (31%) and red kites (22.6%) were exposed and concentrations were higher in North Rhine-Westphalia as well. Among seven MPs, we detected ciprofloxacin (3.4%), which indicates that risk mitigation measures may be needed as resistance genes were already detected in wildlife from Germany. Taken together, our study demonstrates that raptors are exposed to various chemicals during an early life stage depending on their sampling location and underpins that red kites are at particular risk for multiple pesticide exposures in Germany.

Exposure of passerine birds to brodifacoum during management of Norway rats on farms.

The exposure of non-target wildlife to anticoagulant compounds used for rodent control is a well-known phenomenon. Exposure can be primary when non-target species consume bait or secondary via uptake of poisoned animals by mammalian and avian predators. However, nothing is known about the exposure patterns in passerine birds that are commonly present on farms where rodent control is conducted. We used liquid chromatography coupled with tandem mass spectrometry to screen for residues of anticoagulant rodenticides (ARs) in liver tissue of passerine birds that were present during rodent control with a product containing brodifacoum (BR). The 222 birds of 13 species were bycatch of rodent snap trapping in 2011-2013 on 11 livestock farms run synchronously with baiting. During baiting, ARs were detected in about 30% of birds; 28% carried BR. In liver tissue of 54 birds that carried BR, concentrations ranged from 4 to 7809 ng/g (mean 490 ± 169 ng/g). Among common bird species with AR residues, BR was most prevalent in robins (Erithacus rubecula) (44%) and dunnocks (Prunella modularis) (41%). Mean BR concentration was highest in great tits (Parus major) (902 ± 405 ng/g). The occurrence and concentrations of BR residues were about 30% higher in birds collected close to bait stations compared to birds collected further away. The results demonstrate that several ground feeding songbird species are exposed to ARs used on farms. If BR was present in liver tissue, concentrations were variable, which may imply a combination of primary and secondary exposure of songbirds. Exposure was mostly restricted to the immediate surroundings of farms where bait was used, which might limit the transfer to the wider environment. Efforts should be made to reduce the access for birds to AR bait to prevent high exposure.

Baiting location affects anticoagulant rodenticide exposure of non-target small mammals on farms.

BACKGROUND: Commensal rodents such as Norway rats (Rattus norvegicus Berk.), black rats (R. rattus L.) and house mice (Mus musculus L.) damage stored produce and infrastructure, cause hygienic problems and transmit zoonotic pathogens to humans. The management of commensal rodents relies mainly on the use of anticoagulant rodenticides (ARs). ARs are persistent and bio-accumulative, which can cause exposure of non-target species. We compared the baiting strategies to use brodifacoum (BR) in bait boxes indoors only versus in and around buildings in replicated field trials at livestock farms to assess resulting BR residues in non-target small mammals. RESULTS: When bait was used indoors only, the percentage of trapped non-target small mammals with BR residues as well as BR concentration in liver tissue was about 50% lower in comparison to bait application in and around buildings. These effects occurred in murid rodents and shrews but not in voles that were generally only mildly exposed. During the baiting period, BR concentration in murids was stable but decreased by about 50% in shrews. CONCLUSION: Restricting the application of BR bait to indoors only can reduce exposure of non-target species. The positive effect of this baiting strategy on non-target species needs to be balanced with the need for an effective pest rodent management within a reasonable time. More research is needed to clarify which management approaches strike this balance best.

Effects of anticoagulant rodenticide poisoning on spatial behavior of farm dwelling Norway rats.

Commensal rodent species cause damage to crops and stored products, they transmit pathogens to people, livestock and pets and threaten native flora and fauna. To minimize such adverse effects, commensal rodents are predominantly managed with anticoagulant rodenticides (AR) that can be transferred along the food chain. We tested the effect of the uptake of the AR brodifacoum (BR) by Norway rats (Rattus norvegicus) on spatial behavior because this helps to assess the availability of dead rats and residual BR to predators and scavengers. BR was delivered by oral gavage or free-fed bait presented in bait stations. Rats were radio-collared to monitor spatial behavior. BR residues in rat liver tissue were analyzed using liquid chromatography coupled with tandem mass spectrometry. Norway rats that had consumed BR decreased distances moved and had reduced home range size. Treatment effects on spatial behavior seemed to set in rapidly. However, there was no effect on habitat preference. Ninety-two percent of rats that succumbed to BR died in well-hidden locations, where removal by scavenging birds and large mammalian scavengers is unlikely. Rats that ingested bait from bait stations had 65% higher residue concentrations than rats that died from dosing with two-fold LD50. This suggests an overdosing in rats that are managed with 0.0025% BR. None of the 70 BR-loaded rats was caught/removed by wild predators/scavengers before collection of carcasses within 5-29 h. Therefore, and because almost all dead rats died in well-hidden locations, they do not seem to pose a significant risk of AR exposure to large predators/scavengers at livestock farms. Exposure of large predators may originate from AR-poisoned non-target small mammals. The few rats that died in the open are accessible and should be removed in routine searches during and after the application of AR bait to minimize transfer of AR into the wider environment.

Two-year field data on neonicotinoid concentrations in guttation drops of seed treated maize (Zea mays).

We present neonicotinoid concentrations in guttation drops of commonly used maize (Zea mays) cultivars, germinated from seeds coated with active substances (a.s.): i) imidacloprid (IMD), ii) clothianidin (CTN) and iii) thiamethoxam (THM) over two growing seasons. In one variant clothianidin was applied as seed granule. The trial took place at the experimental fields of the Julius Kühn-Institut in Berlin in 2010 and 2011. Data from 2010 are related to a presentation of "Pesticides in guttation droplets following seed treatment - field studies" (Schenke et al., 2011) [1] presented at the SETAC North America conference and only some figures were used in the "Scientific opinion on the science behind the development of a risk assessement of plant protection products on bees (Apis mellifera, Bombus spp. and solitary bees)" (EFSA, 2012) [2]. Only parts of the data from 2011 was presented in relation to the "Exposure of Coccinellidae to guttation droplets on maize seedlings with seed or granule treatment of neonicotinoids" (Schenke and Heimbach, 2014) [3]. The article describes the study sites, the variants of treated maize seeds, sample collection and the analytical methods used to quantify the neonicotinoids and relevant metabolites of IMD (5-OH-IMD and IMD-olefine) and of THM (CTN) in guttation drop samples. The complete field data set is publicly available at the OpenAgrar repository under https://doi.org/10.5073/20180907-142020 (Schenke et al., 2018) [4].

Prey composition modulates exposure risk to anticoagulant rodenticides in a sentinel predator, the barn owl.

Worldwide, small rodents are main prey items for many mammalian and avian predators. Some rodent species have pest potential and are managed with anticoagulant rodenticides (ARs). ARs are consumed by target and non-target small mammals and can lead to secondary exposure of predators. The development of appropriate risk mitigation strategies is important and requires detailed knowledge of AR residue pathways. From July 2011 to October 2013 we collected 2397 regurgitated barn owl (Tyto alba) pellets to analyze diet composition of owls on livestock farms in western Germany. 256 of them were fresh pellets that were collected during brodifacoum baiting. Fresh pellets and 742 liver samples of small mammals that were trapped during baiting in the same area were analyzed for residues of ARs. We calculated exposure risk of barn owls to ARs by comparing seasonal diet composition of owls with AR residue patterns in prey species. Risk was highest in autumn, when barn owls increasingly preyed on Apodemus that regularly showed AR residues, sometimes at high concentrations. The major prey species (Microtus spp.) that was consumed most frequently in summer had less potential to contribute to secondary poisoning of owls. There was no effect of AR application on prey composition. We rarely detected ARs in pellets (2 of 256 samples) but 13% of 38 prey individuals in barn owl nests were AR positive and substantiated the expected pathway. AR residues were present in 55% of 11 barn owl carcasses. Fluctuation in non-target small mammal abundance and differences in AR residue exposure patterns in prey species drives exposure risk for barn owls and probably other predators of small mammals. Exposure risk could be minimized through spatial and temporal adaption of AR applications (avoiding long baiting and non-target hot spots at farms) and through selective bait access for target animals.

Relation between Intensity of Biocide Practice and Residues of Anticoagulant Rodenticides in Red Foxes (Vulpes vulpes).

Anticoagulant rodenticides (ARs) are commonly used to control rodent infestations for biocidal and plant protection purposes. This can lead to AR exposure of non-target small mammals and their predators, which is known from several regions of the world. However, drivers of exposure variation are usually not known. To identify environmental drivers of AR exposure in non-targets we analyzed 331 liver samples of red foxes (Vulpes vulpes) for residues of eight ARs and used local parameters (percentage of urban area and livestock density) to test for associations to residue occurrence. 59.8% of samples collected across Germany contained at least one rodenticide, in 20.2% of cases at levels at which biological effects are suspected. Second generation anticoagulants (mainly brodifacoum and bromadiolone) occurred more often than first generation anticoagulants. Local livestock density and the percentage of urban area were good indicators for AR residue occurrence. There was a positive association between pooled ARs and brodifacoum occurrence with livestock density as well as of pooled ARs, brodifacoum and difenacoum occurrence with the percentage of urban area on administrative district level. Pig holding drove associations of livestock density to AR residue occurrence in foxes. Therefore, risk mitigation strategies should focus on areas of high pig density and on highly urbanized areas to minimize non-target risk.

Spatial and temporal exposure patterns in non-target small mammals during brodifacoum rat control.

Worldwide pest rodents on livestock farms are often regulated using anticoagulant rodenticides (ARs). Second generation ARs in particular can cause poisoning in non-target species due to their high toxicity and persistence. However, research on exposure of small mammals is rare. We systematically investigated spatial and temporal exposure patterns of non-target small mammals in a large-scale replicated study. Small mammals were trapped at different distances to bait stations on ten farms before, during and after brodifacoum (BR) bait application, and liver samples of 1178 non-target small mammals were analyzed for residues of eight ARs using liquid chromatography coupled with tandem mass spectrometry. BR residues were present in 23% out of 742 samples collected during and after baiting. We found clear spatial and temporal exposure patterns. High BR residue concentrations mainly occurred within 15m from bait stations. Occurrence and concentrations of residues significantly decreased with increasing distance. This pattern was found in almost all investigated taxa. After baiting, significantly more individuals contained residues than during baiting but concentrations were considerably lower. Residue occurrence and concentrations differed significantly among taxa, with the highest maximal residue concentrations in Apodemus species, which are protected in Germany. Although Sorex species are known to be insectivorous we regularly found residues in this genus. Residues of active agents other than brodifacoum were rare in all samples. The confirmation of substantial primary exposure in non-target small mammals close to the baiting area indicates considerable risk of secondary poisoning of predators, a pathway that was possibly underestimated until now. Our results will help to develop risk mitigation strategies to reduce risk for non-target small mammals, as well as their predators, in relation to biocidal AR usage.

Elevated root retention of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in coniferous trees.

For decades, the explosive RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) has been used for military and industrial applications. Residues of RDX pollute soils in large areas globally and the persistence and high soil mobility of these residues can lead to leaching into groundwater. Dendroremediation, i.e. the long-term use of trees to clean up polluted soils, is gaining acceptance as a green and sustainable strategy. Although the coniferous tree species Norway spruce and Scots pine cover large areas of military land in Central Europe, the potential of any coniferous tree for dendroremediation of RDX is still unknown. In this study, uptake experiments with a (14)C-labelled RDX solution (30 mg L(-1)) revealed that RDX was predominantly retained in the roots of 6-year-old coniferous trees. Only 23 % (pine) to 34 % (spruce) of RDX equivalents (RDXeq) taken up by the roots were translocated to aboveground tree compartments. This finding contrasts with the high aerial accumulation of RDXeq (up to 95 %) in the mass balances of all other plant species. Belowground retention of RDXeq is relatively stable in fine root fractions, since water leaching from tissue homogenates was less than 5 %. However, remobilisation from milled coarse roots and tree stubs reached up to 53 %. Leaching from homogenised aerial tree material was found to reach 64 % for needles, 58 % for stems and twigs and 40 % for spring sprouts. Leaching of RDX by precipitation increases the risk for undesired re-entry into the soil. However, it also opens the opportunity for microbial mineralisation in the litter layer or in the rhizosphere of coniferous forests and offers a chance for repeated uptake of RDX by the tree roots.