A non-invasive genetic method to identify the sympatric mustelids pine marten (<Emphasis Type="Italic">Martes martes</Emphasis>) and stone marten (<Emphasis Type="Italic">Martes foina</Emphasis>): preliminary distribution survey on the northern Iberian Peninsula

The closely related mustelids European pine marten (Martes martes) and stone marten (Martes foina) sympatrically inhabit a large area of Europe. However, given our limited knowledge of their bioecological relationships, their extremely elusive behaviour and the fact that their faeces cannot be distinguished on the basis of morphology alone, it is very difficult to monitor their populations. In this study, we describe a reliable non-invasive polymerase chain reaction (PCR)–restriction fragment length polymorphism (PCR-RFLP) method for distinguishing between M. martes and M. foina based on the analysis of deoxyribonucleic acid extracted from faeces samples. The method was specifically designed to avoid possible interference from potential prey mammals and other sympatric carnivores. The procedure consists of PCR amplifying a mitochondrial D-loop region followed by digesting the resulting 276-bp-long amplicons with the restriction enzymes HaeIII and RsaI. To assess the efficiency of this technique, we conducted a preliminary field study across the potential sympatric distribution areas of both marten species in the northern Iberian Peninsula. Out of 359 faeces samples collected, we identified 80 as specimens from the stone marten and 235 from the pine marten. Unequivocal species identification was thus possible in 88% of the faeces samples collected. These findings reveal the combined use of non-invasive genetic sampling and GIS technology to be a reliable and cost-effective procedure for improving our knowledge of the spatial distributions of sympatric marten species. This protocol could also be used to identify and improve information gaps, to develop effective research and management programmes and in population and landscape genetics studies of marten species. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

First record of Skrjabingylus petrowi (Nematode: Metastrongyloidea) in a pine marten Martes martes from Ireland

We report the first record of the parasitic nematode Skrjabingylus petrowi in Ireland, the fifth record of S. petrowi outside Russia. Twenty-three of the nematodes were removed from the skull of a male road kill pine marten Martes martes from Co. Tipperary. The spicule lengths measured in the male nematodes removed ranged from 410 to 500 μm, which is in agreement with the range of 410 to 605 μm previously recorded for S. petrowi. This investigation presents further evidence that anatomical measurements are required for the accurate diagnosis of Skrjabingylus spp. and that S. petrowi has a host preference for Martes spp.     

Efficient species identification of pine marten (<Emphasis Type="Italic">Martes martes</Emphasis>) and red fox (<Emphasis Type="Italic">Vulpes vulpes</Emphasis>) scats using a 5′ nuclease real-time PCR assay

Monitoring wildlife species by DNA identification of samples collected non-invasively is an important tool in conservation management. DNA identification of species from faecal (scat) samples is problematic due to the small quantities and poor quality of the DNA isolated from such samples. This study demonstrates the use of real-time PCR technology in the identification of red fox (Vulpes vulpes) and pine marten (Martes martes). It is shown that real-time PCR can be used to identify fox and pine marten by either melting curve analysis (Tm determination) with SYBR Green 1 detection or by the use of species specific fluorogenic probes. The technique is shown to work efficiently with scat DNA.     

A PCR-RFLP-based method to distinguish sable (<Emphasis Type="Italic">Martes zibellina</Emphasis>) and pine marten (<Emphasis Type="Italic">Martes martes</Emphasis>)

Species identification is an important issue in conservation and a particular focus for wildlife forensics. Molecular biological methods retain a unique power to differentiate between difficult samples that lack other identifiable characteristics. The pine marten (Martes martes) and sable (Martes zibellina) are closely related species with very similar pelage characteristics and are often difficult to distinguish from each other. The sable, however, in contrast to the pine marten, remains an endangered and protected animal in China with both hunting and fur trade strictly prohibited for this species. Here, we present a polymerase chain reaction-based restriction fragment length polymorphism method for distinguishing the two species. We sequenced a 638-bp fragment of cytochrome b gene in 39 sables, 68 pine martens, and 10 stone martens and identified all variable nucleotides. A new primer pair was subsequently designed to amplify a 316-bp fragment containing restriction sites of enzyme BseG I and BamH I that are different among martens. When the fragment was cut using BseG I, the resulting restriction pattern was identical in the sable and pine marten, but differed from all other martens. When cut using BamH I, the fragment generated two diagnostic fragments in the sable which could distinguish them from pine martens. This method was valid for all haplotypes of sable and pine marten thus far identified and has high potentially applicability for the identification of the two species.     

Habitat requirements of Eurasian pine marten <Emphasis Type="Italic">Martes martes</Emphasis> in a Mediterranean environment

Evaluating presence and habitat requirements of small carnivores is essential for their conservation. The Eurasian pine marten Martes martes, often described as a habitat specialist associated primarily with forest habitats, has been recently found to live even in patchily wooded country and in shrublands. We evaluated the environmental factors that determine the distribution of the pine marten in a Mediterranean landscape on the island of Sardinia (central Italy). Camera trapping sessions and scat surveys were carried out to assess the presence of the species, then a potential distribution model was developed using ecological niche factor analysis (ENFA), which requires only presence data. The pine marten selected highest altitudes, shrublands, rocky areas, and woodlands, and avoided urban areas and arable lands. Our results indicate that pine marten distribution in our study area is constrained by these variables. The ENFA analysis provided important clues about the distribution range of M. martes and its preferential environmental conditions, updating knowledge of its ecological requirements in Italy.     

Food habits of genetically identified pine marten (<Emphasis Type=Italic>Martes martes</Emphasis>) expanding in agricultural lowlands (NW Italy)

We assessed the diet of pine marten (Martes martes) expanding in the heavily human-altered agricultural plain of the River Po, northern Italy. Between February 2008 and November 2009, surveys were carried out twice a week during seasonal sampling sessions of three to four consecutive weeks. To distinguish the faecal samples from those of sympatric carnivores, a polymerase chain reaction-restriction fragment length polymorphism method was applied. The availability of small mammals was assessed by the analysis of 59 barn owl (Tyto alba) pellets. A total of 109 pine marten faeces were analysed. Its diet consisted of fruit, rodents, lagomorphs and birds. Seasonal variation occurred for fruit, which prevailed in summer, and rodents, which were more preyed upon in autumn. In winter, the diet of the pine marten was almost totally based on vertebrates, and lagomorphs were the main source of protein in summer. Use of small mammals differed significantly from their availability, voles, particularly bank vole Myodes glareolus, being preferred to mice (Apodemus sp.) and rats (Rattus sp.). Medium-size mammals formed about 18% of pine marten diet, a value generally reported for high-latitude habitats. In agricultural areas, the pine marten proved to be an opportunistic predator, able to face the reduced availability of small mammals by preying upon medium-size prey and fruit.     

Life cycle of <Emphasis Type="Italic">Aylax hypecoi</Emphasis> (<Emphasis Type="Italic">Insecta: Hymenoptera: Cynipidae</Emphasis>), a gall inducer on <Emphasis Type="Italic">Hypecoum</Emphasis> ssp. (<Emphasis Type="Italic">Papaveraceae</Emphasis>)

The life cycle and developmental stages of Aylax hypecoi (Trotter, 1913, Hymenoptera: Cynipidae: Aylacini) were studied in detail. Aylax hypecoi is known to induce galls in fruits of two Hypecoum species — H. imberbe and H. geslini (Papaveraceae) and the larva develops in host plant fruits. The morphology and development of egg, larva and pupa were investigated, which has previously not been done. The shape and size of terminal-instar larvae and associated galls are sex-specific. Overwintering stage, adult emergence and flying periods, and egg productivity were studied also.     

Non-invasively determined multi-site variation in pine marten <Emphasis Type="Italic">Martes martes</Emphasis> density,a recovering carnivore in Europe

Pine martens (Martes martes) are subject to national and international conservation legislation throughout most of their European range. Yet population density and abundance estimates, which are a key component of effective conservation management, are lacking in many countries. In this study, a large-scale non-invasive survey was undertaken in 14 forested study sites throughout Ireland to assess variation in pine marten density and abundance. Pine marten hair samples were collected in each study site and analysed using genetic techniques to determine individual identity data. Density and abundance estimates were obtained using spatially explicit capture-recapture models and CAPWIRE. Across all study sites, a total of 93 individual pine marten were identified and captured 217 times. Estimated pine marten density varied from 0 to 2.60 individuals per km² of forested habitat, with all but a single site having estimated densities of ≤1 pine marten per km² of forest habitat. Mean population abundance estimates across all study sites ranged from 0 to 27 individuals. Spatially explicit capture-recapture models on combined data across all 14 study sites provided a mean density estimate of 0.64 (95% CI 0.49–0.81). Combining this with data on the current distribution and estimated area of forest habitat occupied by the species in Ireland, the total pine marten population abundance of pine marten in Ireland was estimated at 3043 (95% CI 2330–3852) individuals. This research has conducted the largest scale investigation of pine marten density and abundance in any part of its global distribution and provided an improved basis for future population assessment and monitoring of this species.     

Lure sticks as a method to detect pine martens <Emphasis Type=Italic>Martes martes</Emphasis>

Marten species are usually surveyed by trapping, snow tracking or cameratrapping with baits on trees. While testing the efficiency of a monitoring scheme for wildcats Felis silvestris silvestris in north-western Switzerland, we noticed that martens are attracted by lure sticks scented with valerian. On these sticks, the animals leaved some hairs that allowed us to identify the genus Martes by microscopic analysis. Additionally, the animal can be identified on pictures made by a camera trap posed close to the lure stick. In this paper, we compared the efficiency of different methods to find the most appropriate one in order to survey the pine marten Martes martes (Linnaeus, 1758) in Switzerland. For the method of valerian lure sticks we estimated a detectability of 0.08 per 14 days during the whole year. The detectability raised, when we applied American Hawbaker’s marten lure instead of the valerian tincture. In addition, the detectability was higher during the period April to June (p=0.2) compared to the whole year. If we identified the pine marten on the lure stick with pictures from the camera traps we reached a detectability of 0.21 during the whole year. Using only camera traps with baits on trees we could not take any picture of a pine marten.     

A species distribution model for pine marten (<Emphasis Type="Italic">Martes martes</Emphasis>) in the least forested region of Europe

Species distribution modelling is a useful technique that provides data on factors that can influence a species range, identify high suitability areas and model future scenarios. The pine marten (Martes martes) has undergone major historical declines in abundance and distribution in Northern Ireland, similar to that which has occurred throughout its range. Currently, the species is in a phase of range expansion in Northern Ireland, in what is the least forested landscape in Europe. To assess the suitability of this environment for pine marten re-establishment, presence only distribution data combined with landcover data at a 10-km scale were used in a species distribution modelling study using Maxent. The results indicated that approximately 32% (4500 km²) of the land area of Northern Ireland had a high probability of pine marten occurrence. Pine marten distribution was positively associated with the extent of conifer forest landcover types, which also had the highest single attribute contribution to the model. Landcover types that were negatively associated with pine marten distribution included the extent of open, dwarf and urban areas.     

<Emphasis Type="Italic">Bacillus anthracis</Emphasis>, <Emphasis Type="Italic">Francisella tularensis</Emphasis> and <Emphasis Type="Italic">Yersinia pestis</Emphasis>. The most important bacterial warfare agents — <Emphasis Type="Italic">review</Emphasis>

There are three most important bacterial causative agents of serious infections that could be misused for warfare purposes: Bacillus anthracis (the causative agent of anthrax) is the most frequently mentioned one; however, Fracisella tularensis (causing tularemia) and Yersinia pestis (the causative agent of plague) are further bacterial agents enlisted by Centers for Disease Control and Prevention into the category A of potential biological weapons. This review intends to summarize basic information about these bacterial agents. Military aspects of their pathogenesis and the detection techniques suitable for field use are discussed.     

Synthesis of orthogonally protected (<Emphasis Type="Italic">3R</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)- and (<Emphasis Type="Italic">3S</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)-4,5-diamino-3-hydroxypentanoic acids

Summary.  The paper describes two methods of the synthesis of ethyl (3R,4S)- and (3S,4S)-4-[(benzyloxycarbonyl)amino]-5-[(tert-butyloxycarbonyl)amino]-3-hydroxypentanoates, useful for the syntheses of edeine analogs. Differently N-protected (S)-2,3-diaminopropanoic acid was used as a substrate in both procedures. The absolute configuration of newly generated asymmetric carbon atoms C-3 in β-hydroxy-γ,δ-diamino products was assigned by means of ¹H NMR spectroscopy after their transformation into corresponding piperidin-2-ones. Received May 24, 2002 Accepted October 10, 2002 Published online December 18, 2002 Acknowledgment The authors are indebted to the Faculty of Chemistry, Technical University of Gdańsk for financial support. Authors' address: Zbigniew Czajgucki, M. Sc., Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Technical University of Gdańsk, 11/12 Narutowicza St., 80-952 Gdańsk, Poland, Fax +48 58 347 11 44, E-mail: zmczaj@wp.pl     

Genetic analysis of sable (<Emphasis Type=Italic>Martes zibellina</Emphasis>) and pine marten (<Emphasis Type=Italic>M. martes</Emphasis>) populations in sympatric part of distribution area in the northern Urals

Analysis of nucleotide sequences of the mitochondrial DNA (mtDNA) control region (495 bp) of sables (Martes zibellina) and pine martens (M. martes) from allopatric parts of the species ranges has shown a considerable interspecific genetic distance (>3%). In sympatric populations of these species in the northern Urals, differences between two species-specific mtDNA lineages are still large; however, classification of each individual nucleotide sequence with one of the two lineages is not correlated with whether the given animal is phenotypically a sable, a pine marten, or a potential hybrid (the so-called “kidas”). This indicates a high degree of reciprocal introgression of the sable and pine marten mtDNA in the northern Urals and suggests that their interspecific hybridization is common in the sympatric zone.     

A new name in <Emphasis Type="Italic">Pavetta</Emphasis> L. (<Emphasis Type="Italic">Rubiaceae</Emphasis>)

Summary  The name Pavetta modesta (Hiern) S. E. Dawson is a later homonym of P. modesta Bremek. Pavetta crystalensis is proposed as a new name.