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Despite the successful conservation of the Przewalski’s horse (PH), Equus ferus przewalskii Groves, 1986 in captivity and reintroduction attempts in Mongolia, there is little knowledge on the genetic variability of these populations. We studied 76 non-metric characters of 130 PH skulls to assess epigenetic variability within and epigenetic distance between temporal and geographical populations of the PH, as well as their fluctuating asymmetry. Our assessment includes skull samples of original native, captive, and reintroduced PH populations. We found the epigenetic variability of original (Iev = 0.21) and captive (Iev = 0.22) PH populations to be similar; the epigenetic variability of reintroduced PH populations was slightly higher (Iev = 0.25). We compared the degree of epigenetic distance among original native, captive, and reintroduced PH populations spanning over 110 years. A low degree of fluctuating asymmetry (FA = 0.05 - 0.08) found in all populations we examined indicates no signs of reduced developmental stability. This suggests that the PH populations are not under undue environmental stress, and there is no evidence of effects of inbreeding depression.

Additional file 1: Table S1. Pearson’s correlations among all weather variables measured during the research. Table S2. Pearson’s correlations among all the weather variables and the percentage representation of each studied behavioural categories.

With increasing livestock numbers, competition and avoidance are increasingly shaping resource availability for wild ungulates. Shifts in the dietary niche of wild ungulates are likely and can be expected to negatively affect their fitness. The Mongolian Gobi constitutes the largest remaining refuge for several threatened ungulates, but unprecedentedly high livestock numbers are sparking growing concerns over rangeland health and impacts on threatened ungulates like the Asiatic wild ass (khulan). Previous stable isotope analysis of khulan tail hair from the Dzungarian Gobi suggested that they graze in summer but switch to a poorer mixed C3 grass / C4 shrub diet in winter, most likely in reaction to local herders and their livestock. Here we attempt to validate these findings with a different methodology, DNA metabarcoding. Further, we extend the scope of the original study to the South Gobi Region, where we expect higher proportions of low-quality browse in the khulan winter diet due to a higher human and livestock presence. Barcoding confirmed the assumptions behind the seasonal diet change observed in the Dzungarian Gobi isotope data, and new isotope analysis revealed a strong seasonal pattern and higher C4 plant intake in the South Gobi Region, in line with our expectations. However, DNA barcoding revealed C4 domination of winter diet was due to C4 grasses (rather than shrubs) for the South Gobi Region. Slight climatic differences result in regional shifts in the occurrence of C3 and C4 grasses and shrubs, which do not allow for an isotopic separation along the grazer-browser continuum over the entire Gobi. Our findings do not allow us to confirm human impacts upon dietary preferences in khulan as we lack seasonal samples from the South Gobi Region. However, these data provide novel insight into khulan diet, raise new questions about plant availability versus preference, and provide a cautionary tale about indirect analysis methods if used in isolation or extrapolated to the landscape level. Good concordance between relative read abundance of C4 genera from barcoding and proportion of C4 plants from isotope analysis adds to a growing body of evidence that barcoding is a promising quantitative tool to understand resource partitioning in ungulates.

In climates with seasonally limited precipitation, terrestrial animals congregate at high densities at scarce water sources. We hypothesize that viruses can exploit the recurrence of these diverse animal congrega- tions to spread. In this study, we test the central prediction of this hypothesis — that viruses employing this transmission strategy remain stable and infectious in water. Equid herpesviruses (EHVs) were cho- sen as a model as they have been shown to remain stable and infectious in water for weeks under labo- ratory conditions. Using fecal data from wild equids from a previous study, we establish that EHVs are shed more frequently by their hosts during the dry season, increasing the probability of water source contamination with EHV. We document the presence of several strains of EHVs present in high genome copy number from the surface water and sediments of waterholes sampled across a variety of mamma- lian assemblages, locations, temperatures and pH. Phylogenetic analysis reveals that the different EHV strains found exhibit little divergence despite representing ancient lineages. We employed molecular approaches to show that EHVs shed remain stable in waterholes with detection decreasing with increas- ing temperature in sediments. Infectivity experiments using cell culture reveals that EHVs remain infectious in water derived from waterholes. The results are supportive of water as an abiotic viral vector for EHV

Department of Zoology, School of Biology and Biotechnology, National University of Mongolia, University Avenue-1, Ulaanbaatar,Mongolia†Mongolia Program, Wildlife Conservation Society, Amar Street-3, “Internom” Bookstore Building, 3rd Floor, Ulaanbaatar, Mongolia‡Ochir-WWF Mongolia Program, Amar Street-4, P.O. Box 115, PO-20A, Ulaanbaatar, Mongolia§The Nature Conservancy Mongolia Program Office, Amar Street-4, 2nd floor, Ulaanbaatar, Mongolia¶Protected Areas Administration, Mongolia, Great Gobi B Strictly Protected Area, Khovd, Mongolia

For certain remote areas like Mongolia, field-based precipitation, surface and ground water isotopic data are scarce. So far no such data exist for the Mongolian Gobi desert, which hinders the understanding of isotopic fractionation processes in this extreme, arid region. We collected 26 event-based precipitation samples, 39 Bij river samples, and 75 samples from other water bodies in the Dzungarian Gobi in SW Mongolia over a period of 16 months for hydrogen and oxygen stable isotope analysis. δ 2H and δ 18O values in precipitation show high seasonal variation and cover an extreme range: 175 ‰ for δ 2H and 24 ‰ for δ 18O values. The calculated local meteoric water line (LMWL) shows the isotopic characteristics of precipitation in an arid region. Individual water samples fall into one of three groups: within, above or below the 95 % confidence interval of LMWL. Data presented provide a basis for future studies in this region.

For certain remote areas like Mongolia, field-based precipitation, surface and ground water isotopic data are scarce. So far no such data exist for the Mongolian Gobi desert, which hinders the understanding of isotopic fractionation processes in this extreme, arid region. We collected 26 event-based precipitation samples, 39 Bij river samples, and 75 samples from other water bodies in the Dzungarian Gobi in SW Mongolia over a period of 16 months for hydrogen and oxygen stable isotope analysis. δ2H and δ18O values in precipitation show high seasonal variation and cover an extreme range: 175 ‰ for δ2H and 24 ‰ for δ18O values. The calculated local meteoric water line (LMWL) shows the isotopic characteristics of precipitation in an arid region. Individual water samples fall into one of three groups: within, above or below the 95 % confidence interval of LMWL. Data presented provide a basis for future studies in this region.

Accurately estimating abundance of wildlife is critical for establishing effective conservation and management strategies. Aerial methodologies for estimating abundance are common in developed countries, but they are often impractical for remote areas of developing countries where many of the world’s endangered and threatened fauna exist. The alternative terrestrial methodologies can be constrained by limitations on access, technology, and human resources, and have rarely been comprehensively conducted for large terrestrial mammals at landscape scales. We attempted to overcome these problems by incorporating local peoples into a simultaneous point count of Asiatic wild ass (Equus hemionus) and goitered gazelle (Gazella subgutturosa) across the Great Gobi B Strictly Protected Area, Mongolia. Paired observers collected abundance and covariate metrics at 50 observation points and we estimated population sizes using distance sampling theory, but also assessed individual observer error to examine potential bias introduced by the large number of minimally trained observers. We estimated 5671 (95% CI = 3611–8907) wild asses and 5909 (95% CI = 3762–9279) gazelle inhabited the 11,027 km 2 study area at the time of our survey and found that the methodology developed was robust at absorbing the logistical challenges and wide range of observer abilities. This initiative serves as a functional model for estimating terrestrial wildlife abundance while integrating local people into scientific and conservation projects. This, in turn, creates vested interest in conservation by the people who are most influential in, and most affected by, the outcomes.

Kaczensky, P., Enkhsaikhan, N., Ganbaatar, O. & Walzer, C. 2008: The Great Gobi B Strictly Protected Area in Mongolia - refuge or sink for wolves Canis lupus in the Gobi? - Wildl. Biol. 14: 444-456. The Mongolian hunting law does not mention the wolf Canis lupus, which is generally interpreted in the way that wolves can be hunted anytime and anywhere, including in protected areas. We investigated whether the Great Gobi B Strictly Protected Area (SPA), a strict nature reserve in southwestern Mongolia, acts as a refuge or a sink for wolves in the Gobi. Our expectations were that wolves in the Gobi 1) have large ranges similar to those in other equally unproductive habitats, 2) experience a high hunting pressure, and 3) have recently become an important export item for cross-border trade to China. We combined GPS positions of two adult wolves, wolf harvest data and a market survey on wildlife products to address the above questions. Range use of the two collared wolves was huge, but varied widely between the two animals (6,670 km 2 for an adult female and 26,619 km 2 for an adult male) and over time. Reproductive status and residency status were only known during the initial 8-months monitoring period of the female. During this 'resident' period her range size was 1,275 km 2 . Both wolves showed a clear preference of mountainous terrain over flat steppe, suggesting that only 21% of the SPA constitute preferred wolf habitat. Annual harvest in the park and its vicinity averaged 1 wolf/265 km 2 in 2002/03, 1 wolf/120 km 2 in 2003/04 and 1 wolf/310 km 2 in 2004/05. However, hunting pressure was unequally distributed and particularly high in the northeastern corner of the park. During the active monitoring period of wolf F1, 35 wolves were killed within her 'resident' range, suggesting a high hunting pressure. Most wolves were shot from motorised vehicles, possibly explaining the preference of wolves for mountainous terrain which is inaccessible for vehicles. The market surveys revealed products from y2,000 wolves on the two border markets, a huge discrepancy to only 150 CITES permits officially issued annually. Although our data are insufficient to allow a truly quantitative assessment of the impact of human induced mortality on wolf conservation status in the Great Gobi B SPA, it points towards a potentially severe conservation problem requiring further attention.

1. Historically, the overlap zones of wild equids were small in Africa but extensive for Przewalski's horses Equus ferus przewalskii and Asiatic wild asses Equus hemionus in Asia. Currently, the Great Gobi B Strictly Protected Area in south-western Mongolia is the only place where sympatric, free-ranging populations of these equids occur. This provides a unique opportunity to test the hypothesis that Przewalski's horses are primarily adapted to mesic steppes and Asiatic wild asses to arid desert steppes and semi-deserts. Understanding the spatial needs and habitat requirements of these little-studied species is a pre-requisite for setting aside and managing protected areas and planning future re-introductions. 2. From 2001 to 2005, we followed nine Przewalski's horses and seven Asiatic wild asses using satellite telemetry and direct observations to assess differences in their resource selection strategies and social organization. 3. Przewalski's horses had non-exclusive home ranges of 152-826 km 2, selected for the most productive plant communities and formed stable harems groups. 4. Asiatic wild asses had non-exclusive home ranges of 4449-6835 km 2, showed little preferences for any plant community and seemed to live in fission-fusion groups. 5. Synthesis and applications. Our results provide evidence for different resource selection strategies in two sympatric equid species. Our findings indicate that the Gobi areas provide an edge, rather than an optimal habitat for Przewalski's horses. Consequently, only small and isolated pockets of suitable habitat remain for future re-introductions. Asiatic wild asses, on the other hand, need access to large tracts of land to cope with the unpredictable resource distribution of the Gobi. Managers should be aware that protecting habitat where Asiatic wild asses occur does not necessarily benefit Przewalski's horse restoration, whereas setting aside habitat for the conservation of Przewalski's horses will only locally benefit Asiatic wild asses.

Rationale In recent years, segmental stable isotope analysis of hair has been a focus of research in animal dietary ecology and migration. To correctly assign tail hair segments to seasons or even Julian dates, information on tail hair growth rates is a key parameter, but is lacking for most species. Methods We (a) reviewed the literature on tail hair growth rates in mammals; b) made own measurements of three captive equid species; (c) measured δ2H, δ13C and δ15N values in sequentially cut tail hairs of three sympatric, free-ranging equids from the Mongolian Gobi, using isotope ratio mass spectrometry (IRMS); and (d) collected environmental background data on seasonal variation by measuring δ2H values in precipitation by IRMS and by compiling pasture productivity measured by remote sensing via the normalized difference vegetation index (NDVI). Results Tail hair growth rates showed significant inter- and intra-specific variation making temporal alignment problematic. In the Mongolian Gobi, high seasonal variation of δ2H values in precipitation results in winter lows and summer highs of δ2H values of available water sources. In water-dependent equids, this seasonality is reflected in the isotope signatures of sequentially cut tails hairs. Conclusions In regions which are subject to strong seasonal patterns we suggest identifying key isotopes which show strong seasonal variation in the environment and can be expected to be reflected in the animal tissue. The known interval between the maxima and minima of these isotope values can then be used to correctly temporally align the segmental stable isotope signature for each individual animal. © 2015 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.

The Przewalski's horse (Equus caballus przewalskii) was extinct in the wild by the mid 1960s. The species has survived because of captive breeding only. The Takhin Tal reintroduction project is run by the International Takhi Group; it is one of two projects reintroducing horses to the wild in Mongolia. In 1997 the first harem group was released. The first foals were successfully raised in the wild in 1999. Currently, 63 Przewalski's horses live in Takhin Tal. Little information exists on causes of mortality before the implementation of a disease-monitoring program in 1998. Since 1999, all dead horses recovered (n = 28) have been examined and samples collected and submitted for further investigation. Equine piroplasmosis, a tick-transmitted disease caused by Babesia caballi or Theileria equi, is endemic in Takhin Tal and was identified as the cause of death of four stallions and one stillborn foal. In December 2000, wolf predation was implicated in the loss of several Przewalski's horses. However, thorough clinical, pathologic, and bacteriologic investigations performed on dead and surviving horses of this group revealed lesions compatible with strangles. The extreme Mongolian winter of 2000-2001 is thought to have most probably weakened the horses, making them more susceptible to opportunistic infection and subsequent wolf predation. Other occasional causes of death since 1999 were trauma, exhaustion, wasting, urolithiasis, pneumonia, abortion, and stillbirth. The pathologic examination of the Przewalski's horses did not result in a definitive diagnosis in each case. Several disease factors were found to be important in the initial phase of the reintroduction, which could potentially jeopardize the establishment of a self-sustaining population.

The Przewalski’s horse (Equus caballus przewalskii) became extinct in the wild during the 1960s. Based on a successful captive breeding program, Przewalski’s horses were reintroduced to the Great Gobi Part “B” strictly protected area (SPA) in SW Mongolia in the late 1990s. The Asiatic wild ass (Equus hemionus hemionus), Przewalski’s horse, and sometimes domestic horses live sympatricly in the Gobi B SPA. Previously published data demonstrates that, as a result of their different requirements and utilization of the park’s resources, their home-range size and social structure differ. Parasitological examinations in the three equid species show how the factors “home range, social structure, and resource selection” significantly impact parasitic burden. Asiatic wild asses are potentially exposed to a higher risk of parasite re-infection due to their temporal aggregation in very large groups. This study demonstrates a highly significant greater parasite load in the Asiatic wild ass for the majority of parasites evaluated (Dictyocaulus arnfieldi, Trichostrongylus axei, Strongyloides westeri, Parascaris equorum) compared to Przewalski’s horses and domestic horses in the same habitat. Domestic horses had higher parasite loads for eggs of strongylids, eggs of anoplocephalidae, and Eimeria leuckarti. The potential risk of cross infection between sympatric living equids is high, as is the cross infection between ruminants and equids. Furthermore, this study reports for the first time the occurrence of lungworms in free-ranging Przewalski’s horses. Whereas, Asiatic wild asses and Przewalski’s horses seem to cope very well with the sometimes high parasite burden, Mongolian domestic horses manifested typical parasite burden symptoms.