Seroprevalence and risk factors of Toxoplasma gondii infection in invasive raccoons (Procyon lotor) in Central Europe.

Toxoplasma gondii is an obligate intracellular protozoan that causes toxoplasmosis in warm-blooded animals. Most mammals, including humans, can become intermediate host, resulting in subclinical infection or even death. Generally, there is limited information on the epidemiology of T. gondii of game species in Germany. As omnivores, raccoons, which are particularly widespread and abundant in Germany, are particularly exposed to infection the parasite. Here, we report the seroprevalence of T. gondii antibodies from 15 study sites located in Luxembourg and Germany. Using the indirect modified agglutination test (MAT), 170 (37.4%; 95% CI: 33.0-41.9) out of 454 raccoons were surveyed to be T. gondii seropositive. While values ranged from 19.0% to 53.3%, there was no significant difference in seroprevalence between study areas. Animal weight had a strong influence on the presence of T. gondii antibodies in raccoon sera, with heavier animals more likely to be seropositive. Our results show that T. gondii infection is widespread in central European raccoons, suggesting a high degree of ecosystem circulation of the parasite.

Determinants and effects of sinus worm Skrjabingylus nasicola (Nematoda: Metastrongyloidae) infestation in invasive American mink Neovison vison in Germany.

Skrjabingylus nasicola (Leuckart, 1842) are geographically widespread nematodes that parasitize the nasal and frontal sinus cavities of smaller mustelids. As most prior work was solely based on the analysis of bone injuries of museum skull, little is known about the determinants and effects of infestation in the host species. Working on fresh skulls, we aimed to analyze infestation patterns in American mink (Neovison vison Schreber, 1777) from nine study areas in northern Germany and to identify factors that explained infestation prevalence and intensity in the host species. The prevalence (46.7-62.9 %) and infestation intensity values (4.5-10.89 nematodes) reported here were relatively large, especially compared to other American mink populations in Europe. Considering mink diet, our study sites probably harbored a larger number of infested paratenic hosts and climate did not have a substantial negative influence on survival of S. nasicola larvae. We did not observe any significant sex-age differences in either prevalence or intensity of S. nasicola infestation. We did not find a negative impact of an infestation on the host animals' body weight, confirming prior results that the parasite is not a significant mortality factor in mustelids. Our study suggests that this holds even outside the native distributional range where the host's defenses might not be optimally adapted to an autochthonous parasite.

Prevalence and molecular identification of the sinus worm Skrjabingylus petrowi (Nematoda: Metastrongyloidea) from Martes spp. in Germany.

The nematodes of the genus Skrjabingylus (family Metastrongylidae) can parasitise the nasal and frontal sinus cavities of different carnivore species. Until recently, Skrjabingylus petrowi Bageanov & Petrov, 1941, has mainly been described in pine martens (Martes martes Linnaeus, 1758) and sables (Martes zibellina Linnaeus, 1758) sampled in the European part of the former Soviet Union. Newer finds in the stone marten (Martes foina Erxleben, 1777) and from different parts of Europe suggest, however, that the species might have a broader host-species range than previously assumed and be geographically more widespread as well. Since most S. petrowi records have resulted from chance discoveries rather than systematic surveys, very little is known about the prevalence of S. petrowi in marten populations. Here, we report results of a 20-year extensive survey of fresh marten skulls, where we tested 1.059 marten carcasses originating from 248 localities in Germany for the presence of S. petrowi. We identified an infestation in only four M. martes individuals and one M. foina, despite using a reliable identification method. Based on the spicule lengths of the male nematodes, the parasites were identified as S. petrowi and genetic barcoding confirmed the identification of the samples. In a phylogenetic analysis, S. petrowi and Skrjabingylus nasicola (Leuckart, 1842), formed a sister clade to all the other members of the family Metastrongylidae. The low prevalence of S. petrowi is possibly due to its parasitising in the two marten species that are either not very common (M. martes) or predominantly live in urban habitat (M. foina).

Dissections of fresh skulls confirm low prevalence of Troglotrema acutum (Trematoda: Troglotrematidae) in German badgers (Meles meles).

We examined 131 European badgers Meles meles (Linnaeus, 1758) from 67 localities in central Germany for the presence of the cranial trematode Troglotrema acutum, as previous studies based on museum skulls might have underestimated the prevalence of the parasite in this host. We detected the flatworm in only three individuals that originated from the Rhoen Mountains (Thurigina and Bavaria). While the cranium of one host individual showed the lesions and the sponge-like widening of certain regions of the skullcap that are typical of a T. acutum infection, the skulls of the two remaining badgers did not show any deformations. The three badgers were infected by eight, 20, and 49 T. acutum individuals, respectively. Eggs of the trematode parasite were detected in the paranasal sinuses of two badgers. While badgers infected with T. acutum may not show any surface bone lesions, the results of the present study do not contradict the conclusion that the badger is only an accidental host of T. acutum.

Revisiting the phylogeography and demography of European badgers (Meles meles) based on broad sampling, multiple markers and simulations.

Although the phylogeography of European mammals has been extensively investigated since the 1990s, many studies were limited in terms of sampling distribution, the number of molecular markers used and the analytical techniques employed, frequently leading to incomplete postglacial recolonisation scenarios. The broad-scale genetic structure of the European badger (Meles meles) is of interest as it may result from historic restriction to glacial refugia and/or recent anthropogenic impact. However, previous studies were based mostly on samples from western Europe, making it difficult to draw robust conclusions about the location of refugia, patterns of postglacial expansion and recent demography. In the present study, continent-wide sampling and analyses with multiple markers provided evidence for two glacial refugia (Iberia and southeast Europe) that contributed to the genetic variation observed in badgers in Europe today. Approximate Bayesian computation provided support for a colonisation of Scandinavia from both Iberian and southeastern refugia. In the whole of Europe, we observed a decline in genetic diversity with increasing latitude, suggesting that the reduced diversity in the peripheral populations resulted from a postglacial expansion processes. Although MSVAR v.1.3 also provided evidence for recent genetic bottlenecks in some of these peripheral populations, the simulations performed to estimate the method's power to correctly infer the past demography of our empirical populations suggested that the timing and severity of bottlenecks could not be established with certainty. We urge caution against trying to relate demographic declines inferred using MSVAR with particular historic or climatological events.

Population structure of the mosquito Aedes aegypti (Stegomyia aegypti) in Pakistan.

Eleven microsatellite markers were used to determine the genetic population structure and spread of Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) in Pakistan using mosquitoes collected from 13 different cities. There is a single genetic cluster of Ae. aegypti in Pakistan with a pattern of isolation by distance within the population. The low level of isolation by distance suggests the long-range passive dispersal of this mosquito, which may be facilitated by the tyre trade in Pakistan. A decrease in genetic diversity from south to north suggests a recent spread of this mosquito from Karachi. A strong negative correlation between genetic distance and the quality of road connections shows that populations in cities connected by better road networks are less differentiated, which suggests the human-aided passive dispersal of Ae. aegypti in Pakistan. Dispersal on a large spatial scale may facilitate the strategy of introducing transgenic Ae. aegypti or intracellular bacteria such as Wolbachia to control the spread of dengue disease in Pakistan, but it also emphasizes the need for simple measures to control container breeding sites.

Comparative landscape genetic analyses show a Belgian motorway to be a gene flow barrier for red deer (Cervus elaphus), but not wild boars (Sus scrofa).

While motorways are often assumed to influence the movement behaviour of large mammals, there are surprisingly few studies that show an influence of these linear structures on the genetic make-up of wild ungulate populations. Here, we analyse the spatial genetic structure of red deer (Cervus elaphus) and wild boars (Sus scrofa) along a stretch of motorway in the Walloon part of Belgium. Altogether, 876 red deer were genotyped at 13 microsatellite loci, and 325 wild boars at 14 loci. In the case of the red deer, different genetic clustering tools identified two genetic subpopulations whose borders matched the motorway well. Conversely, no genetic structure was identified in the case of the wild boar. Analysis of isolation-by-distance patterns of pairs of individuals on the same side and on different sides of the motorway also suggested that the road was a barrier to red deer, but not to wild boar movement. While telemetry studies seem to confirm that red deer are more affected by motorways than wild boar, the red deer sample size was also much larger than that of the wild boars. We therefore repeated the analysis of genetic structure in the red deer with randomly sub-sampled data sets of decreasing size. The power to detect the genetic structure using clustering methods decreased with decreasing sample size.

Using isolation-by-distance-based approaches to assess the barrier effect of linear landscape elements on badger (Meles meles) dispersal.

As the European badger (Meles meles) can be of conservation or management concern, it is important to have a good understanding of the species' dispersal ability. In particular, knowledge of landscape elements that affect dispersal can contribute to devising effective management strategies. However, the standard approach of using Bayesian clustering methods to correlate genetic discontinuities with landscape elements cannot easily be applied to this problem, as badger populations are often characterized by a strong confounding isolation-by-distance (IBD) pattern. We therefore developed a two-step method that compares the location of pairs of related badgers relative to a putative barrier and utilizes the expected spatial genetic structure characterized by IBD as a null model to test for the presence of a barrier. If a linear feature disrupts dispersal, the IBD pattern characterising pairs of individuals located on different sides of a putative barrier should differ significantly from the pattern obtained with pairs of individuals located on the same side. We used our new approach to assess the impact of rivers and roads of different sizes on badger dispersal in western England. We show that a large, wide river represented a barrier to badger dispersal and found evidence that a motorway may also restrict badger movement. Conversely, we did not find any evidence for small rivers and roads interfering with badger movement. One of the advantages of our approach is that potentially it can detect features that disrupt gene flow locally, without necessarily creating distinct identifiable genetic units.

Using genetic methods to investigate dispersal in two badger (Meles meles) populations with different ecological characteristics.

Understanding the dispersal behaviour of a species is important for understanding its ecology and evolution. Dispersal in the Eurasian badger (Meles meles) is believed to be very limited, with social groups forming primarily through the retention of offspring. However, most of our knowledge of dispersal in this species comes from studies of high-density populations in the United Kingdom, where badgers are atypical in their behaviour, physiology, ecology and prey specialization. In this study we use genetic methods to compare dispersal patterns in a British and a Swiss population that differ in their ecology and demography. We present well-supported evidence that badgers disperse much further in the low-density continental population, where dispersal may also be female biased. Limited dispersal thus seems not to be an intrinsic behavioural characteristic of the species. Rather, dispersal patterns seem to vary depending on population demography and, ultimately, habitat quality and characteristics. This could have important management consequences, as dispersal can affect the impact of local extinction, and host dispersal has a particularly important role in disease transmission. Even though concentrated studies of a species in a single location may not provide representative data for the species, there are few mammalian studies that compare demography and dispersal patterns across contrasting habitats. Our results provide an example of phenotypic plasticity and suggest that dispersal is determined by the interaction of individual, social and environmental factors that may differ between populations.

Genetic structure and assignment tests demonstrate illegal translocation of red deer (Cervus elaphus) into a continuous population.

Molecular forensic methods are being increasingly used to help enforce wildlife conservation laws. Using multilocus genotyping, illegal translocation of an animal can be demonstrated by excluding all potential source populations as an individual's population of origin. Here, we illustrate how this approach can be applied to a large continuous population by defining the population genetic structure and excluding suspect animals from each identified cluster. We aimed to test the hypothesis that recreational hunters had illegally introduced a group of red deer into a hunting area in Luxembourg. Reference samples were collected over a large area in order to test the possibility that the suspect individuals might be recent immigrants. Due to isolation-by-distance relationships in the data set, inferring the number of genetic clusters using Bayesian methods was not straightforward. Biologically meaningful clusters were only obtained by simultaneously analysing spatial and genetic information using the program baps 4.1. We inferred the presence of three genetic clusters in the study region. Using partial Mantel tests, we detected barriers to gene flow other than distance, probably created by a combination of urban areas, motorways and a river valley used for viticulture. The four focal animals could be excluded with a high certainty from the three genetic subpopulations and it was therefore likely that they had been released illegally.

Reliable microsatellite genotyping of the Eurasian badger (Meles meles) using faecal DNA.

The potential link between badgers and bovine tuberculosis has made it vital to develop accurate techniques to census badgers. Here we investigate the potential of using genetic profiles obtained from faecal DNA as a basis for population size estimation. After trialling several methods we obtained a high amplification success rate (89%) by storing faeces in 70% ethanol and using the guanidine thiocyanate/silica method for extraction. Using 70% ethanol as a storage agent had the advantage of it being an antiseptic. In order to obtain reliable genotypes with fewer amplification reactions than the standard multiple-tubes approach, we devised a comparative approach in which genetic profiles were compared and replication directed at similar, but not identical, genotypes. This modified method achieved a reduction in polymerase chain reactions comparable with the maximum-likelihood model when just using reliability criteria, and was slightly better when using reliability criteria with the additional proviso that alleles must be observed twice to be considered reliable. Our comparative approach would be best suited for studies that include multiple faeces from each individual. We utilized our approach in a well-studied population of badgers from which individuals had been sampled and reliable genotypes obtained. In a study of 53 faeces sampled from three social groups over 10 days, we found that direct enumeration could not be used to estimate population size, but that the application of mark-recapture models has the potential to provide more accurate results.

Abdominal wall Actinomyces abscess associated with an intrauterine device. A case report.

An abdominal wall Actinomyces abscess occurred in a woman with an intrauterine device. Contributing factors were local trauma, spread from surrounding colonized body sites and symbiotic growth of other anaerobes. The diagnosis was based on the histologic finding of the sulfur granule. Special studies may be needed to distinguish this condition from other, similar ones (Nocardia, botryomycosis).