An extended mtDNA phylogeography for the alpine newt illuminates the provenance of introduced populations

Many herpetofauna species have been introduced outside of their native range. MtDNA barcoding is regularly used to determine the provenance of such populations. The alpine newt has been introduced across the Netherlands, the United Kingdom and Ireland. However, geographical mtDNA structure across the natural range of the alpine newt is still incompletely understood and certain regions are severely undersampled. We collect mtDNA sequence data of over seven hundred individuals, from both the native and the introduced range. The main new insights from our extended mtDNA phylogeography are that 1) haplotypes from Spain do not form a reciprocally monophyletic clade, but are nested inside the mtDNA clade that covers western and eastern Europe; and 2) haplotypes from the northwest Balkans form a monophyletic clade together with those from the Southern Carpathians and Apuseni Mountains. We also home in on the regions where the distinct mtDNA clades meet in nature. We show that four out of the seven distinct mtDNA clades that comprise the alpine newt are implicated in the introductions in the Netherlands, United Kingdom and Ireland. In several introduced localities, two distinct mtDNA clades co-occur. As these mtDNA clades presumably represent cryptic species, we urge that the extent of genetic admixture between them is assessed from genome-wide nuclear DNA markers. We mobilized a large number of citizen scientists in this project to support the collection of DNA samples by skin swabbing and underscore the effectiveness of this sampling technique for mtDNA barcoding

Cuora mouhotii (Gray 1862) – Keeled Box Turtle

– The Keeled Box Turtle, Cuora mouhotii (Family Geoemydidae), is a small (carapace length up to ca. 250 mm), once poorly-known terrestrial species. Recent extensive research on the species in its natural habitat in northern Vietnam and Hainan, China, has significantly increased available knowledge of the species. Cuora mouhotii is a highly terrestrial turtle from the mesic forests of Southeast Asia. Two subspecies have been described, and intergradient populations are known. Relatively widespread, it is apparently abundant nowhere. Omnivorous, at least in captivity, the species is highly carnivorous in the wild but will also take fruit. Clutch size ranges from 1–9 eggs, with egg dimensions ca. 41 x 26 mm; incubation period in captivity varies from 82 to 120 days. Major threats to the species are loss of forest habitat from logging and commercial trade for Traditional Chinese Medicine and food. To a lesser extent capture for local consumption and both national and international pet trade are factors threatening this species

An isolated crested newt population in Dutch coastal dunes: distribution relict or introduction?

Isolated distribution patches may represent local remnants of a formerly wider range or could have originated by human-mediated expansion beyond the natural range. Distinguishing between these two scenarios is not always straightforward. Northern crested newts (Triturus cristatus) in the Dutch coastal dunes are disconnected from the main species range by over 40 kilometres and whether they have been present historically is unclear. We genotyped crested newts from throughout the Netherlands for an mtDNA marker to determine the provenance of the coastal dune population. Because a closely related species, the Italian crested newt (T. carnifex), has an introduction history in the Netherlands, we also screened eight nuclear DNA SNP markers diagnostic for T. cristatus vs. T. carnifex. The crested newts from the coastal dunes carry a single T. cristatus mtDNA haplotype that naturally occurs in the south, but not the east, of the Netherlands. Therefore, we cannot distinguish if the population represents a natural distribution relict or is derived from an introduction. We find no evidence of genetic admixture with T. carnifex in the coastal dunes, but such admixture is apparent at another Dutch locality (far removed from a previously known genetically admixed population). Our study illustrates how difficult it can be to determine the origin of isolated populations. </p

The conservation paradox of an introduced population of a threatened species: spadefoot toads in the coastal dunes of the Netherlands

   Species that are threatened in their native range, may actually prosper as introduced populations. To investigate how such introduced populations were established involves determining from where within the natural range the founder individuals originated. This can be accomplished through mtDNA barcoding. The common spadefoot toad (Pelobates fuscus) naturally occurs in the south and east of the Netherlands and has shown a rapid decline. Yet, a flourishing introduced population was recently discovered in the coastal dunes in the west of the country. We use mtDNA barcoding to determine the provenance of the introduced population. We sampled both native and introduced populations from the Netherlands and compared our sequences to haplotypes from across the entire distribution range. The mtDNA haplotypes found in the introduced population are distinct from those naturally occurring in the Netherlands and point towards an origin in the Pannonian Basin, on the boundary between Central and Southeastern Europe. Paradoxically, the thriving P. fuscus population in the Dutch coastal dunes should be considered a conservation risk to local biodiversity, even though within the native range in the Netherlands the species is severely threatened. Our study illustrates the complicated conservation questions associated with species that are both native and invasive.</p

Course of an Isolated Ranavirus Outbreak in a Pelobates fuscus Population in The Netherlands

Viruses in the genus Ranavirus (family Iridoviridae) are known to have the potential to adversely affect fish, amphibians, and reptiles. Ranaviruses are associated with large-scale die-offs and rapid population declines in amphibians. The development and progression of an outbreak, however, vary greatly depending on the host species and geographic location. We describe the recurrent course of an outbreak of common midwife toad virus in an isolated population (Staphorst) of common spadefoot toads (Pelobates fuscus) in The Netherlands from 2012 to 2015. After initial mass mortality of toad larvae in 2012, no mass mortality was recorded in 2013 and 2014. In 2015, however, a recurrent outbreak of the virus is believed to have caused high mortality rates among this species in the Staphorst population

The introduction of three cryptic tree frog species in the Dutch coastal dunes challenges conservation paradigms

Invasive species are considered one of the main drivers of the sixth mass extinction. Conservation solutions depend on whether a species is also indigenous to the country it invades (i.e., beyond its native range). In the case of invasive cryptic species, genetic tools are required to establish their identity. We illustrate these issues with the human-mediated colonization of the Dutch coastal dunes by Hyla tree frogs. Although previously assumed to concern the indigenous common tree frog H. arborea, European tree frogs comprise a complex of allopatric cryptic species, meaning the taxonomic identity of introduced Dutch populations warrants investigation. We sequence mtDNA for 164 individuals from native and introduced populations across the Netherlands and compare our dataset with hundreds of Hyla haplotypes previously barcoded in the Western Palearctic. Two of the dune populations carry an mtDNA haplotype of the native species H. arborea that occurs naturally elsewhere in the Netherlands. In contrast, mtDNA assigned to the eastern tree frog H. orientalis was detected in all three other dune populations. In one of these populations mtDNA of the Italian tree frog H. intermedia was also found. Not one, but three species of tree frogs have thus been introduced to the Dutch coastal dunes, only one of them being native to the Netherlands. This situation causes a conservation conundrum as some introduced populations are lawfully protected but could pose a threat to local biodiversity. Regarding the ‘true’ exotic tree frog species, all conservation options should be considered. </p

The limits of mtDNA analysis for determining the provenance of invasive species: a midwife toad example

For most if not all European herpetofauna, range-wide mtDNA phylogeographies have been published. This facilitates establishing the provenance of introduced populations. However, precision is contingent on the spatial genetic structure across the range of the taxon under study and, in particular, from where within that range the introduction was sourced. In the Netherlands, the common midwife toad, Alytes obstetricans, only naturally occurs in the extreme southeast and is on the decline there. Yet, introduced populations thrive elsewhere in the country. We use mtDNA analysis to try to determine the origin of two introduced populations along the Dutch coast, in the city of The Hague and the dune area Meijendel. We compiled a database of hundreds of individuals from throughout the distribution range and added over 130 individuals from both native and introduced populations from the Netherlands, Belgium and Germany. The mtDNA haplotypes found in the introduced populations are associated with postglacial expansion. The main haplotype predominates in the natural range in the Netherlands, but also occurs much more widely across western Europe, north of the Pyrenees. A closely related haplotype, newly identified from The Hague, was not found in the native Netherlands range, suggesting an origin from abroad. The combination of low phylogeographic resolution and low sampling density in the postglacially colonized part of the range hampers our ability to determine the provenance of the introduced A. obstetricans populations. </p

Rarely naturalized, but widespread and even invasive: the paradox of a popular pet terrapin expansion in Eurasia

The North American terrapin, the red-eared slider, has globally recognized invasive status. We built a new extensive database using our own original and literature data on the ecology of this reptile, representing information on 1477 water bodies throughout Eurasia over the last 50 years. The analysis reveals regions of earliest introductions and long-term spatio-temporal dynamics of the expansion covering now 68 Eurasian countries, including eight countries reported here for the first time. We established also long-term trends in terms of numbers of terrapins per aquatic site, habitat occupation, and reproduction success. Our investigation has revealed differences in the ecology of the red-eared slider in different parts of Eurasia. The most prominent expression of diverse signs of invasion success (higher portion of inhabited natural water bodies, higher number of individuals per water body, successful overwintering, occurrence of juvenile individuals, successful reproduction, and establishment of populations) are typical for Europe, West Asia and East Asia and tend to be restricted to coastal regions and islands. Reproduction records coincide well with the predicted potential range based on climatic requirements but records of successful wintering have a wider distribution. This invader provides an excellent and possibly unique (among animals) example of wide alien distribution, without the establishment of reproducing populations, but through the recruitment of new individuals to rising pseudopopulations due to additional releases. Therefore, alongside the potential reproduction range, a cost-effective strategy for population control must take in account the geographical area of successful wintering. Graphical abstrac