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The degree of introgressive hybridization between the Scottish wildcat and domestic cat has long been suspected to be advanced. Here, we use a 35‐SNP‐marker test, designed to assess hybridization between wildcat and domestic cat populations in Scotland, to assess a database of 295 wild‐living and captive cat samples, and test the assumptions of the test using 3,097 SNP markers generated independently in a subset of the data using ddRAD. We discovered that despite increased genetic resolution provided by these methods, wild‐living cats in Scotland show a complete genetic continuum or hybrid swarm structure when judged against reference data. The historical population of wildcats, although hybridized, clearly groups at one end of this continuum, as does the captive population of wildcats. The interpretation of pelage scores against nuclear genetic data continues to be problematic. This is probably because of a breakdown in linkage equilibrium between wildcat pelage genes as the two populations have become increasingly mixed, meaning that pelage score or SNP score alone is poor diagnostic predictors of hybrid status. Until better tools become available, both should be used jointly, where possible, when making management decisions about individual cats. We recommend that the conservation community in Scotland must now define clearly what measures are to be used to diagnose a wildcat in the wild in Scotland, if future conservation action is to be effective.

23Environmental DNA (eDNA) metabarcoding can identify terrestrial taxa utilising aquatic habitats 24 alongside aquatic communities, but terrestrial species' eDNA dynamics are understudied. We 25 evaluated eDNA metabarcoding for monitoring semi-aquatic and terrestrial mammals, 26 specifically nine species of conservation or management concern, and examined 27 spatiotemporal variation in mammal eDNA signals. We hypothesised eDNA signals would be 28 stronger for semi-aquatic than terrestrial mammals, and at sites where individuals exhibited 29 behaviours. In captivity, we sampled waterbodies at points where behaviours were observed 30 ('directed' sampling) and at equidistant intervals along the shoreline ('stratified' sampling). We 31 surveyed natural ponds (N = 6) where focal species were present using stratified water 32 sampling, camera traps, and field signs. eDNA samples were metabarcoded using vertebrate-33 specific primers. All focal species were detected in captivity. eDNA signal strength did not differ 34 between directed and stratified samples across or within species, between semi-aquatic or 35 terrestrial species, or according to behaviours. eDNA was evenly distributed in artificial 36 waterbodies, but unevenly distributed in natural ponds. Survey methods deployed at natural 37 ponds shared three species detections. Metabarcoding missed badger and red fox recorded by 38 cameras and field signs, but detected small mammals these tools overlooked, e.g. water vole. 39Terrestrial mammal eDNA signals were weaker and detected less frequently than semi-aquatic 40 mammal eDNA signals. eDNA metabarcoding could enhance mammal monitoring through 41 large-scale, multi-species distribution assessment for priority and difficult to survey species, and 42 provide early indication of range expansions or contractions. However, eDNA surveys need high 43 3 spatiotemporal resolution and metabarcoding biases require further investigation before 44 routine implementation. 45 46

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. AbstractAim: We explore the phylogeography of Himalayan wolves using multiple genetic markers applied on a landscape-scale dataset and relate our findings to the biogeographic history of the region. Location: Himalayas of Nepal, the Tibetan Plateau of China and mountain ranges of Central Asia. Taxon: Himalayan wolf (also called the Tibetan wolf), Canis lupus chanco. Methods: We present a large-scale, non-invasive study of Himalayan wolves from across their estimated range. We analysed 280 wolf scat samples from western China, Kyrgyzstan and Tajikistan at two mtDNA loci, 17 microsatellite loci, four nonsynonymous SNPs in three nuclear genes related to the hypoxia pathway, and ZF genes on both sex chromosomes. | 1273 WERHAHN Et Al.