On the occurrence of the Himalayan Wolf Canis lupus, L. 1758 (Mammalia: Carnivora: Canidae) in the Gaurishankar Conser- vation Area, Nepal; its existence confirmed through sign and visual evidence in Rolwaling Valley

The Himalayan Wolf Canis lupus L., a top predator of the Third Pole, is proposed to be of a distinct wolf lineage (C. himalayensis) relative to the Holarctic Grey Wolf as described by mtDNA analyses. A biodiversity survey organized by the Gaurishankar Conservation Area Project (GCAP) has captured images of wolves in three different regions, and the study team has observed wolf scats in five additional regions above the tree line in Rolwaling Valley. Further, interviews with local herders provided evidence of wolf depredation of livestock in the area. The Rolwaling Valley in the Gaurishankar Conservation Area was the study area which was divided into 12, 4 x 4 km (16 km2) grid cells, each supplied with one camera trap operated continuously from June to November 2019 (only 6 out of 12 cameras functioned for the duration of our study). Wolf detections were recorded by camera traps from Yalung Pass (4,956 m), Tsho-Rolpa glacial Lake (4,536 m) and the Dudhkunda ridgeline (5,091 m). The photo capture rate index (PCRI) for wolves was 0.71. Our study reports the first photographic evidence of the Himalayan Wolf in the Rolwaling Valley.


INTRODUCTION
The Himalayan Wolf Canis lupus is a top predator of the Third Pole (Prater 1971;Menon 2003;Chetri et al. 2017;Boitani et al. 2018;Werhahn et al. 2020). It was proposed to be a distinct wolf lineage (C. himalayensis; Aggarwal et al. 2007) relative to the Holarctic Grey Wolf as described by mtDNA analyses (Sharma et al. 2004;Chetri et al. 2016;Chetri et al. 2017;Werhahn et al. 2017;Boitani et al. 2018). Categorized as 'Least Concern' by the IUCN (Boitani et al. 2018), it is considered 'Critically Endangered' by the National Red List in Nepal (Jnawali et al. 2011). International trade is generally prohibited by CITES (The Convention on International Trade in Endangered Species of Wild Fauna and Flora) with the Himalayan Wolves in Nepal listed under Annex 1 (listed species are the most endangered species and are threatened with extinction; CITES 2019). Furthermore, capture, killing, and trade are strictly prohibited as well by declaring this species under Schedule-I in Nepal by The National Parks and Wildlife Protection Act, 1973(NLC 1973. In and around the lap of the Himalaya, wolves were recorded by researchers at the Indo-Pakistan Himalaya and also from the Himachal Pradesh and Uttarakhand states of India near the western border of Nepal above 3,500 m (Sharma et al. 2004;Bhattacharya & Sathyakumar 2010;Chetri et al. 2016;Werhahn et al. 2017). In Nepal, the wolf is reported from all the Himalayan protected area systems and adjoining wilderness areas above 4,000 m (Subba et al. 2017). The areas that are considered main wolf habitat in Nepal include Manasalu and Annapurna Conservation Area (Chetri et al. 2016, 2017, Subba et al. 2017, Shey Phoksundo National Park and Humla district of western Nepal (Werhahn et al. 2017;Subba et al. 2017), Dhorpatan Hunting Reserve (Jnawali et al. 2011), andKanchenjungha Conservation Area (Jnawali et al. 2011;Subba et al. 2017); and now its existence has also been confirmed in the Gaurishankar Conservation Area (this study).

Study area
The Gaurishankar Conservation Area (GCA) is one of the newest protected areas of Nepal, covering 2,179 km 2 in the northern part of the Ramechhap, Dolakha, and Sindhupalchok districts of the Bagmati Province of Federal Democratic Republic of Nepal (Bajracharya et al. 2011, NLC 2074. It extends between 85°46.8'- 86°34.8' East & 27°34.2'-28°10' North, and ranges in elevation 980-7,134 m (Bajracharya et al. 2011;GCAP 2013). The GCA was established in 2010 (GoN 2010) connecting two national parks in the high mountains (i.e., Sagarmatha National Park in the east and Langtang National Park in the west) with the northern boundary extending to the Nepal-China border (Bajracharya et al. 2011). The valley is situated in the northeastern corner of the GCA, approximately 50 km west of Mt. Everest (Sacherer 2011).
Our study area included 16 major vegetation types, river valleys of Koshi River basin, snow-capped mountains, and temperate-alpine grasslands supporting 565 species of vascular plants, 76 (71 species described in GCAP, 2013 and five added from this study) species of mammals, 252 species of birds, 12 species of amphibians, 27 reptiles, and 27 species of fishes (Bajracharya et al. 2011;GCAP 2013 Gaurishankar Conservation Area includes two 'beyul' (a Tibetan word meaning sacred) valleys named Lapchi and Rolwaling where the majority of human inhabitants (i.e., the Sherpa community) follow Buddhism where as in other valleys, there is a mosaic of Hindu-Buddhist religion occurring (Bajracharya et al. 2011;Sacherer 2011;GCAP 2013). The Rolwaling valley is sacred landscape guided by Tibetan Buddhism and inhabited by the Sherpa community. Followers of the 'Padmasambhava' sects of Buddhism (Sacherer 2011) strictly obey and maintain a ban on animal sacrifice and consumption of animal meat, and people in this region believe that consuming meat products and burning of garlic brings misfortune (Lama 2019). Thus, wild animals are not harvested for their meat, providing some measure of wildlife protection. Traditionally, the valley dwellers rely on the agro-pastoral economy with transhumance animal herding, the primary cause of human-wolf negative interaction in this valley. Although, the killing of animals is prohibited, the last known wolf pack in this area was poisoned by Yak herders ~50 years ago (Sherpa 2019).
The Rolwaling Valley (215 km 2 ) covers the landscapes of lower temperate forests up to the alpine zone. Major vegetation types of the study area were Quercus forests, lower-temperate mixed broadleaved forests, upper J TT temperate broadleaved forests, rhododendron forests, and upper temperate conifer forests (Bajracharya, et al. 2011). The higher areas are covered by Abies-Juniper forests and birch-rhododendron forests, whereas the alpine zone comprises alpine shrub land, scrubland, open grassland, glaciers, and rocky outcrops (Bajracharya et al. 2011;GCAP 2013

Blocks and grids
We divided the GCA into five blocks as defined by geographical barriers and ease of research management and we selected the Rolwaling Valley block for this study. A recent research objective in this valley was to assess mammalian diversity, with grids created to target Snow Leopard detection (Jackson et al. 2005). The valley covers 215 km 2 with accessible areas divided into 16 km 2 square grids ( Figure 1) following Jackson et al. (2005). We focused our monitoring efforts on those grids above 3,000 m targeting large mammals J TT of the upper temperate-alpine region with low human habitation (Bajimaya 2000). Altogether, we placed one camera in each of the12 grids that were 3,000-5,500 m in elevation.

Camera trapping
In the current study, the target species were the elusive species of the highlands including the Snow Leopard, Wolf, Lynx and their prey species. Camera trapping was selected as the primary method given feasibility and logistical challenges. (i.e., limited resources precluded transect and genetic surveys).
The study team tried to minimize disturbance to wildlife throughout the survey, although livestock herders traveled periodically to the survey area. Cameras were placed far from major trekking routes and the primary trails used by herders were omitted for camera security reasons. This also minimized the chance of capturing images of non-target animals (livestock) and humans. Cameras were left unattended for an extended period (around six months), during which the possibility of wildlife occurrence was estimated to be the highest based on various signs and marks of animals.
Bushnell Trophy Camera Brown (Model 119436) and Bushnell Trophy Camera w/viewscreen (Model 119455) were used for monitoring animals. Cameras operated continuously for six months between 09 June and 09 November 2019. Cameras were set to image capture mode with one minute lag between triggers with three images captured per trigger. No fixed camera height was applied as per the objective of the study but cameras were set to focus around 30 cm height over the trail i.e. some cameras were tilted to achieve the specified focal height.
We identified sites of likely animal movement within each grid by visually examining the site characteristics. The sites with high frequency of signs of animal like tracks, scat deposits, pellets, rubbing on trees, scent marks, and trail junctions were selected for camera installment, following the Snow Leopard monitoring manual for Nepal (Bajimaya 2000). Moreover, valley bottoms and ridgelines, where the likelihood of megafauna movement is high (Jackson et al. 2005), were also selected for camera deployment. No baits or trail modifications were used.

Data analysis
Photocapturerate index (PCRI) is used as an index of animal abundance because of its general relationship with the density of target species (Rovero & Marshall 2009: Lahkar et al. 2018. Although, its application is better suited to the prey species (Rovero & Marshall 2009;Lahkar et al. 2018), we applied it to predators as well because of limited data available for capture recapture analysis and because individual identification of wolves was not reliable.
We calculated total operation time summing up data from all cameras that were functional (6 out of 12) (i.e., six cameras were not functional: four cameras triggered continuously resulting in filled SD cards within a couple of days, rain water leakage damaged the storage device in one camera, and one camera was lost during this study). Every photo event was recorded by a photo analysis using a digital projector. To define a photographic event, 30 minutes between events of same species was used to assure independent data points.

Additional lines of evidence
Wolf scats were opportunistically observed and recorded during other field work. Also, interviews were conducted with local herders to obtain information on wolf depredation of livestock in the area.

RESULTS AND DISCUSSION
The PCRIs for mammals and birds were calculated separately (Table 1). Among mammals, Pika was the most frequently photographed, followed by cattle (Yak), Red Fox, Yellow-bellied Weasel, and Wolf (0.71 animals per 100 nights, i.e., 0.71 PCRI values) (Table 1). Ungulate prey species that are frequently observed along mountain slopes, such as the Himalayan Tahr, Common Goral, and the Himalayan Serow, had lower PCRI values. However, game birds such as Blood Pheasant, Himalayan Snow Cock, and Himalayan Monal, were frequently photographed (Table 1).
Based on the photographic evidence, all wolf detections consisted of single wolves, no packs or pairs were detected in the six captures of wolf ( Figure  2). Sniffing on scent sprays and travel were common behaviors observed. The movements of wolf were recorded by three cameras located at 5,091 m, 4,536 m, and 4,956 m in the Rolwaling Valley. This is the first ever photographic record of wolf presence, not only in Rolwaling Valley, but also in the GCA.

Scat observation and depredation history
Scats of wolves were observed during transect walks for finding suitable camera sites (Figure 3). Identifiable scats were observed over 3,900 m on Rhododendron anthopogon dominated scrublands, on the human/ domestic animal tracks. Also, tracks were observed in moraines. Recently, a couple of livestock depredation events near Naa village of the valley were recorded. In both occasions herders had managed to chase down small packs of Wolves (Lama 2019; Sherpa 2019). Table  2 and Figure 2 show the sites where signs of Wolf were found and general site characteristics. Earlier, a reconnaissance survey carried out by GCAP (2016) also observed Wolf scat in the Numbur Valley (behind Yalung Peak).
Although, the presence of the wolf in GCA was reported based on interviews and indirect signs (Bajracharya et al. 2011), our study confirms its presence through photographs, scat observations, and information from livestock depredations (Figure 4). This paper reports the first visual proofof the wolf in Rolwaling Valley. To our knowledge, this is probably the first ever photograph of the wolf in the Gaurishankar landscapes visually confirming its re-colonization in the valley.

CONCLUSION
This study provides the first ever photographic evidence confirming the presence of the Himalayan Wolf in the northern area (Rolwaling region) of GCA in Nepal. Six wolf-detection events were obtained by camera trapping, all consisting of single wolves. Also, the five wolf scats and tracks that were observed during camera sites selection and depredation information from local interviewees further confirmed the wolf's presence in this area. The sites, where the evidence of wolf were confirmed, also overlapped with areas used by snow leopard and red foxes. Although, this study was not intended to quantify human-carnivore conflict, interviews confirmed that Snow Leopards are not the only predators in Rolwaling region that accounts for human wildlife interactions, especially in relation to the depredation of domesticated animals such as mountain goat and sheep, as well as Yak and 'Djokpa' (cross breed between Yak and cow). Our study aimed to assess the mega faunal biodiversity in the Rolwaling area. Additional research is warranted, specifically a more in-depth assessment of the status, habitat range, behavior and ecological role of the Wolf. In part because wolves may be contributing to human wildlife conflict in the high Himalayan regions, conservation interventions may be needed to prevent local extinction of this species as a result of human-retaliatory killings. Furthermore, because the taxonomy of this Wolf is being debated and may result in a unique species identified, we suggest a precautionary conservation strategy be developed and implemented. www.threatenedtaxa.org The Journal of Threatened Taxa (JoTT) is dedicated to building evidence for conservation globally by publishing peer-reviewed articles online every month at a reasonably rapid rate at www.threatenedtaxa.org. All articles published in JoTT are registered under Creative Commons Attribution 4.0 International License unless otherwise mentioned. JoTT allows allows unrestricted use, reproduction, and distribution of articles in any medium by providing adequate credit to the author(s) and the source of publication.