**1. Introduction**

To ensure that captive breeding programs are more robust and sustainable, it is important to ethically and legally ensure optimum animal welfare [1]. The rapid developmental processes of humans are a ffecting the natural habitats of wildlife. Thus, wildlife reservoirs, zoos, and enclosures must be adapted to minimize the e ffect of these changes [2]. Today, captive breeding is one of the most important conservation tools [3], providing an opportunity to the rare endangered species to produce stable populations for eventual release into the wild [4].

In the meantime, enclosures and other facilities for wild animals are under severe pressure to limit animals to small areas [5]. Thus, animal welfare subjects, particularly those related to the captive wild species, are rapidly recognized [6,7]. It is important to ensure the optimum levels of animal welfare of captive animals for the production and maintenance of healthy, viable populations [8]. To make captive breeding more robust, it is important to determine the main factors of species welfare and, most importantly, the welfare of every individual of a particular group [9].

Animal welfare assessment protocols can be designed by gathering information through simple inspections, animal observations, and visiting animal facilities and enclosures [10]. Animal welfare assessment protocols for livestock (poultry, cattle, and pigs) have already been developed under the auspices of the Welfare Quality ® project. These protocols are mainly based on animal-based indicators, in addition to also having resource or environmental measures [11]. Animal-based measures can be directly recorded by observing animals, including their physical appearance, health, and behavior. Unlike animal-based measures, environmental measures assess the available resources for these animals in captivity, and the animal itself is not taken into account.

*Ovis orientalis* (urial) is a wild sheep that closely resembles Marco Polo sheep in general body appearance [12]. In Pakistan, the urial is represented by three subspecies: *Ovis vignei vigeni* (Ladakh urial), which is restricted to northern areas (Gilgit-Baltistan) of Pakistan; *Ovis vignei punjabiensis* (Punjab urial) found in the Salt Range (Punjab) and the northwestern province of Khyber Pakhtunkhwa (formerly known as the North-West Frontier Province); and *Ovis vignei blanfordi* (Baluchi urial), which is found in the southwestern province of Balochistan [13]. The Punjab urial is a gregarious ungulate, and most big herds include females, lambs, and immature males. It has been observed that the urial generally prefers grasses, but can also be found foraging on shrubs [14].

The species has been declared as vulnerable globally, with a declining population trend, according to the International Union for Conservation (IUCN) list of threatened species [15], and is endangered in Pakistan [16–18]. In Pakistan, wild ungulates are reared in captivity, but there are no standardized methods and protocols to measure the welfare of these captive ungulates. Therefore, this current study aimed to (i) design and develop a baseline welfare assessment protocol for captive Punjab urial based on the livestock welfare assessment protocol from the Welfare Quality ® project, (ii) implement this welfare assessment protocol in facilities hosting Punjab urial, and (iii) sugges<sup>t</sup> recommendations for better captive breeding and management.

#### **2. Materials and Methods**

For this study, we selected the subspecies *Ovis orientalis punjabiensis* because of its availability in captivity. The current study was conducted in two steps. In the first step, the welfare assessment protocol was developed, and, in the second step, the newly established protocol was implemented at captive facilities housing Punjab urial. We developed the welfare assessment protocol by combining results from other studies on the biology and behavior of the species and the sheep welfare assessment protocol [19], as both domestic sheep and Punjab urial belong to the family Bovidae.

To obtain information on the general biology and behavior of the species, we used Google Scholar and Web of Science search engines using "Ovis vignei" as keywords. Limited scientific published information is available regarding the biology of this species in natural habitats. Moreover, no work has been conducted to investigate the problems faced by this species in captivity. More than 31 scientific published papers were reviewed. Most of these published work focused on population status, population dynamics, diet ecology, and habitat, of which two papers [12,20] provided useful detailed information on the behavior and general biology of the species that was utilized for the welfare assessment protocol.

For developing the Punjab urial welfare assessment protocol, four basic principles were taken into account: good feeding, good housing, good health, and suitable behavior [10,11]. These principles led to twelve criteria (see Section 3), which in turn allowed the development of welfare assessment indicators [21]. After combining information from [19,22], we developed an extensive set of 31 welfare indicators for Punjab urial (see Section 3).

The final version of the welfare assessment protocol was then applied to three different groups of captive Punjab urial at two different facilities—Cherat Wildlife Park (CWP) in Nowshera and Manglot Wildlife Park (MWP) in Nizampur—in the month of August 2019. Both of these parks are located in the Nowshera District of Khyber Pakhtunkhwa Province, Pakistan. The captive breeding program was launched in 2008 at CWP with a single pair of founder animals, while in 2012 it was initiated at MWP, also with a single pair. The groups of both programs were mixed herds, including adult males, sub adult males, adult females, sub adult females, and lambs. CWP had one group consisting of 23 individuals (*n* = 23) with a mean age of 3.21 ± 2.21 years. At MWP, two groups were present (*n* = 6 and *n* = 8), with mean ages of 3.16 ± 1.57 and 3.33 ± 1.69 years, respectively. Groups were represented by coding their facilities (centers) as captive Punjab urial-1 (CU1) at CWP, and captive Punjab urial-2 (CU2) and captive Punjab urial-3 (CU3) at MWP. The protocol was applied by the same person.
