Hans Dautel

Geographical distribution of Dermacentor marginatus and Dermacentor reticulatus in Europe.

The goal of this paper is to present up-to-date maps depicting the geographical distribution of Dermacentor species in Europe based on georeferenced sampling sites. Therefore, a dataset was compiled, resulting in 1286 D. marginatus (Sulzer, 1776) and 1209 D. reticulatus (Fabricius, 1794) locations. Special emphasis is given to the region of the European Alps depicting a presumable climate barrier of the mountains and to overlaps in the distribution of both species as well as on the situation in eastern European countries. For the latter newly described Dermacentor findings comprise 59 locations in Romania and 62 locations in Ukraine. The geographical distributions of both species in Europe range from Portugal to Ukraine (and continue to the east of Kazakhstan). Although it is well known that D. marginatus is adapted to a warmer and drier climate at more southern latitudes and D. reticulatus to a moderately moist climate at more northern latitudes, the distribution limits of both species were not well known. Here, the northern and southern distribution limits for both species in Europe, as determined from the georeferenced database, were specified for D. marginatus by the belt of 33-51° N latitude and for D. reticulatus by the belt of 41-57° N latitude. Thus, overlapping species distributions were found between 41° N and 51° N.

Species determination and characterization of developmental stages of ticks by whole-animal matrix-assisted laser desorption/ionization mass spectrometry

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of crude bacterial samples has been introduced as a very cost-efficient and rapid, yet highly informative tool to identify and classify bacteria. The potential of this approach to characterize whole animals, so far preferentially insects, is only evolving. Here, a simple protocol was developed to perform MALDI-MS analysis on extracts from whole ticks of 7 species and 4 developmental stages. Using commercially available software designed for the identification of bacteria, a reference database of spectra was constructed that allowed the species determination of ticks using larvae, nymphs, or adult individuals as starting material. Cluster analysis on the basis of MALDI mass spectra indicated that the primary determinant for the mass spectra was the species, followed by the developmental stages, which formed distinct clusters within the given species. With certain limitations, species identification was also possible using body parts and engorged animals. Spectra of developing Ixodes ricinus eggs showed dramatic changes with time, suggesting that, beyond its usefulness for species determination, MALDI-typing may have applications in developmental biology.

A novel test system for detection of tick repellents

A bioassay was developed to examine the response of ticks towards potential repellents that may protect vertebrates against tick bites. Such tick repellents must be effective despite the unavoidable presence of various attractive host‐derived stimuli. Therefore, a moving‐object‐bioassay (MO‐bioassay) was developed that mimicks body warmth and movement of vertebrates by a rotating and heated drum. Compounds which were tested for their effects on ticks were applied onto a small elevated area of the drum. Ticks were allowed to approach the drum by walking on a glass rod which ended ≈1 mm away from the local elevation. Ticks could cling to this elevation that intermittently passed by, whereas the remaining drum surface was too far away from the tip of the rod to be contacted by the ticks.

Prevention of infectious tick-borne diseases in humans: Comparative studies of the repellency of different dodecanoic acid-formulations against Ixodes ricinus ticks (Acari: Ixodidae)

BackgroundTicks of the species Ixodes ricinus are the main vectors of Lyme Borreliosis and Tick-borne Encephalitis – two rapidly emerging diseases in Europe. Repellents provide a practical means of protection against tick bites and can therefore minimize the transmission of tick-borne diseases. We developed and tested seven different dodecanoic acid (DDA)-formulations for their efficacy in repelling host-seeking nymphs of I. ricinus by laboratory screening. The ultimately selected formulation was then used for comparative investigations of commercially available tick repellents in humans.MethodsLaboratory screening tests were performed using the Moving-object (MO) bioassay. All test formulations contained 10% of the naturally occurring active substance DDA and differed only in terms of the quantitative and qualitative composition of inactive ingredients and fragrances. The test procedure used in the human bioassays is a modification of an assay described by the U.S. Environmental Protection Agency and recommended for regulatory affairs. Repellency was computed using the equation: R = 100 - NR/N × 100, where NR is the number of non-repelled ticks, and N is the respective number of control ticks. All investigations were conducted in a controlled laboratory environment offering standardized test conditions.ResultsAll test formulations strongly repelled nymphs of I. ricinus (100-81% protection) as shown by the MO-bioassay. The majority of ticks dropped off the treated surface of the heated rotating drum that served as the attractant (1 mg/cm2 repellent applied). The 10% DDA-based formulation, that produced the best results in laboratory screening, was as effective as the coconut oil-based reference product. The mean protection time of both preparations was generally similar and averaged 8 hours.Repellency investigations in humans showed that the most effective 10% DDA-based formulation (~1.67 mg/cm2 applied) strongly avoided the attachment of I. ricinus nymphs and adults for at least 6 hours. The test repellent always provided protection (83-63%) against I. ricinus nymphs equivalent to the natural coconut oil based reference product and a better protection (88-75%) against adult ticks than the synthetic Icaridin-containing reference repellent.ConclusionWe found that the 10% DDA-based formulation (ContraZeck®) is an easily applied and very effective natural repellent against I. ricinus ticks. By reducing the human-vector contact the product minimises the risk of transmission of tick-borne diseases in humans.

The first German map of georeferenced ixodid tick locations

BackgroundGeoreferenced locations of ixodid ticks are required to depict the observed distribution of species. Further, they are used as input data for species distribution models also known as niche models. The latter were applied to describe current and future (projected) tick distributions. Beside model assumptions and selected climate parameters, the number of georeferenced tick locations available as a digital dataset is of fundamental importance for the reliability of such models. For Germany, however, no comprehensive dataset of ixodid tick species exists. The goal of this study was to put together all the available information on ixodid tick locations in Germany to produce such a digital dataset and to visualize it in a map.FindingsA total of 2,044 georeferenced locations of ixodid ticks in Germany were compiled from two existing datasets (altogether 993 locations) and an extensive literature study (1,051 locations). The resulting digital dataset comprises the following tick species: Ixodes ricinus (1,855 locations), Ixodes apronophorus (1), Ixodes frontalis (1), Ixodes hexagonus (1), Ixodes trianguliceps (4), Dermacentor marginatus (77), Dermacentor reticulatus (96), Haemaphysalis concinna (8) and Hyalomma marginatum (1). The data were used to draw a tick map for Germany, showing I. ricinus occurring in the whole federal territory, while D. marginatus has been restricted to the climatically favoured region of the Rhine valley. Clustered locations of D. reticulatus were also documented in the Rhine valley as well as in Berlin and its vicinity.ConclusionsThe introduced map depicts for the first time the available geographical coordinates of ixodid tick locations in Germany. The digital dataset used to draw the map is provided to the scientific community as a basis for further investigations such as species distribution modelling.

Long-term in vitro cultivation of Borrelia miyamotoi

Borrelia are fastidious bacteria some of which are difficult to grow in vitro. Here, we report a method for successful continuous in vitro cultivation of the emerging pathogen Borrelia miyamotoi. The type and quantity of serum as well as the atmosphere were critical for successful in vitro cultivation. Optimal growth was achieved using 50% pooled human serum and an atmosphere of 6% CO2.

The pigeon tick (Argas reflexus): its biology, ecology, and epidemiological aspects.

The European pigeon tick, Argas reflexus (F.), is in central Europe predominantly an urban pest parasitizing wild and domesticated pigeons, Columba livia. Under certain circumstances, however, it also bites humans, occasionally causing an IgE-mediated type-I allergy. Control of A. reflexus is very difficult because of a number of remarkable morphological, physiological, and behavioural features of the tick. The present study aimed at elucidating the distribution and the frequency of occurrence of A. reflexus in Berlin and its possible vector role for Borrelia burgdorferi sensu lato (s. l.). Buildings reported by occupiers to be infested with A. reflexus were personally examined. In addition, the carrier status of this soft tick for B. burgdorferi (s. l.) was investigated in three German towns by an indirect immunofluorescence assay. A total of 188 Berlin buildings was found to be infested with A. reflexus between 1989 and 1998. Infestations were found in 17 out of the 21 districts, clustering in the inner city. There was only a single Borrelia-positive tick out of 800 sampled in Berlin, Leipzig, and Hannover using the genus specific antibody H9724. The same tick was PCR-negative for B. burgdorferi outer surface protein A. Neither these results nor those of other studies on the occurrence of antibodies against B. burgdorferi in pigeons suggest that field populations of A. reflexus and/or pigeons in central Europe harbour the causative agent of Lyme borreliosis.

A Density Map of the Tick-Borne Encephalitis and Lyme Borreliosis Vector Ixodes ricinus (Acari: Ixodidae) for Germany

Abstract The castor bean tick Ixodes ricinus (L.) is the principal vector for a variety of viral, bacterial, and protozoan pathogens causing a growing public-health issue over the past decades. However, a national density map of I. ricinus is still missing. Here, I. ricinus nymphs in Germany were investigated by compiling a high-resolution map depicting the mean annually accumulated nymphal density, as observed by monthly flagging an area of 100 m2. Input data comprise ticks collected at 69 sampling sites. The model domain covers an area of about 357,000 km2 (regional scale). Two negative binomial regression models were fitted to the data to interpolate the tick densities to unsampled locations using bioclimatic variables and land cover, which were selected according to their significance by the Akaike information criterion (AIC). The default model was fitted to the complete dataset resulting in AIC = 842. An optimized model resulted in a significantly better value of AIC = 732. Tick densities are very low in urban (green) areas. Maximum annual densities up to 1,000 nymphs per 100 m2 are observed in broad-leaved forests. The tick maps were verified by leave-one-out cross-validation. Root mean square errors of RMSE = 137 and RMSE = 126 nymphs per 100 m2 were estimated for the two models, respectively. These errors are of the order of the interannual variation of the tick densities. The compilation of a high-resolution density map of unfed nymphal I. ricinus for Germany provides a novel, nationwide insight into the distribution of an important disease vector.

Repellent efficacy of DEET, Icaridin, and EBAAP against Ixodes ricinus and Ixodes scapularis nymphs (Acari, Ixodidae) ☆

Repellent efficacy of 10% EBAAP (3-[N-butyl-N-acetyl]-aminopropionic acid, ethyl ester) and 10% Icaridin ((2-(2-hydroxyethyl)-1-piperidinecarboxylic acid 1-methylpropyl ester)) were evaluated against 20% DEET (N,N-diethyl-3-methylbenzamide) in human subject trials against ticks. Responses of host-seeking nymphs of the European castor bean tick (Ixodes ricinus L.; Acari: Ixodidae) and the North American blacklegged tick (I. scapularis Say; Acari: Ixodidae) were compared. Tests were carried out according to the US-EPA standard protocol with ethanolic solutions of the active ingredients of repellents being applied to the forearm of 10 volunteers. The upward movement of ticks was monitored until repellent failure taking up to 12.5 h. Application of 20% DEET resulted in median complete protection times (CPT; Kaplan-Meier median) between 4 and 4.5 h, while 10% EBAAP yielded CPTs of 3.5-4h. No significant differences were found between the efficacies of two repellents nor between the two species tested. The median of the CPT of a 10% Icaridin solution was 5h in nymphs of I. scapularis, but 8h in those of I. ricinus (P<0.01). Based on these studies, EBAAP and Icaridin are efficacious alternatives to DEET in their repellent activity against nymphs of the two Ixodes ticks with Icaridin demonstrating particularly promising results against I. ricinus. Future research should investigate whether similar results occur when adult Ixodes ticks or other tick species are tested.

Efficacy testing of several Ixodes ricinus tick repellents: different results with different assays.

In the European Union (EU), tick repellents for humans need to be registered and approved by the authorities in order to be marketed. As there are currently no specific technical guidelines for product evaluation, we compared 3 different test methods: the mechanical moving object bioassay (MOB), and 2 assays involving human volunteers. For the latter, procedures according to the U.S. Environmental Protection Agency (EPA) and the Stiftung Warentest (StiWa), a German consumer care organization, were used. Two repellents, Autan(®) (AU), based on 20% Picaridin [2-(2-hydroxyethyl)-1-piperidinecarboxylic acid-1-methylpropyl ester], and ZeckWeck (ZW), based on 12.5g/100g Citriodiol™ (main compound: p-menthane-3,8-diol) were tested with all 3 assays. Three repellents, Anti Brumm(®) naturelle, based on 20% Citriodiol™ (main compound: p-menthane-3,8-diol), G090141, based on 20% EBAAP (ethyl buthyl acetyl aminopropionate), and G090152, based on 10% decanoic acid (capric acid), which is contained in Zanzarin(®), were tested according to the EPA and the StiWa procedures. The EPA assay indicated a significantly higher repellency of the products AU and G090141 than the StiWa test, but no difference between assays could be detected for the remaining 3 products. Also the corresponding protection times were significantly longer (approximately 4h) when determined according to EPA versus to StiWa for 3 of the products, whilst the difference was insignificant for ZW and G090152. Additionally, significantly lower numbers of ticks initially walked onto the repellent-treated skin when tested according to EPA versus to StiWa in all products except ZW and G090152. Thus, the StiWa protocol appears to pose higher demands on a repellent than the EPA method. Contrary to expectation, the MOB showed the same or even lower product efficacy when compared to the EPA and StiWa tests. Particularly, the percentage of ticks clinging to repellent-treated filter paper was significantly higher than the proportion of ticks walking onto treated skin in the other assays. This could mean that in nature more ticks may probably cling to a human protected by a given repellent than the EPA or the StiWa assay might suggest. Nevertheless, the MOB produced results that are quite similar to the tests involving human volunteers.