**3. Results**

#### *3.1. Development of Welfare Assessment Protocol*

The protocols developed in the current study for welfare assessment of Punjab urial consisted of 4 basic principles, 12 criteria, and 31 indicators (Table 1). Of the 12 criteria of the Welfare Quality® protocol for domestic sheep, some were replaced keeping in view the biology of the species. The species under study is a wild ungulate, thus "lack of minerals" was added to the list of criteria. The current study is a leading step in captive Punjab urial welfare assessment, and will provide a base to investigate more complex and less frequently studied aspects in captive animals.


**Table 1.** Punjab urial welfare quality assessment protocol principles, criteria, and indicators. Animal-based indicators are represented by \*.

#### 3.1.1. Lack of Prolonged Appetite

Since there are no husbandry guidelines for this species, there is no parameter that can be used for the description of this criterion. In many animal welfare assessment protocols, however, body condition is included as an animal-based indicator to measure welfare. Both poor and excessive body conditions can lead to extreme and unexpected results. Poor body conditions can show signs of malnutrition, diseases, or extreme hunger. Poor and weak body conditions can have severe negative effects on the reproduction, behavior, and health of the animals [23].

To indicate the welfare problems, both poor and excessive body conditions can be used (indicator 1.1) [24]. To date, there is no scoring scale for the body condition of Punjab urial; hence, the body condition scoring guidelines for two other wild bovids proposed in [10,24] were followed. Animals were observed from a close distance from behind and the side. Punjab urial was scored as "poor body conditions" if ribs, spine, and pelvis were sharp and prominent, and projected with a concave rump. The animal was scored as "normal body condition" if ribs, pelvis, and spine were difficult to be distinguished from one another, with a flat rump area. The animal was scored as "excessive body conditions" if ribs, spine, and pelvis were not visible because of a thick layer of fat, with a protruding rump area.

#### 3.1.2. Lack of Prolonged Thirst

One of the most important welfare requirements is ad libitum, access to water. Water provision in animal welfare protocol is a resource-based indicator. There should be easy access to water sources for the animals, containing clean and fresh water supplied on a daily basis.

According to the current study, water availability (indicator 2.1), in conjunction with the number of water sources (2.2) and cleanliness of water sources (2.3), should be properly checked. All these indicators were validated in [10,25] for assessing the welfare of captive *Dorcas gazelles* and cows. Water was scored as "dirty" if it was polluted with food wastes or plastics, or was dark green in color with a foul smell. On the contrary, water was scored as "clean" if it was without any contamination or foul smell. Remains of fresh feed were acceptable.

#### 3.1.3. Lack of Minerals

In order to maintain normal physiological functions, wild ungulates usually search for a salt lick. Deficiency of salts can have severe negative e ffects on animal health, appetite, reproduction, and lactation [26]. Since captive animals are confined to limited areas in enclosures, they are more prone to mineral deficiency [27]. The current protocol included two indicators—(3.1) "availability of salt licks" and (3.2) "licking objects"—to check whether the animals were mineral-deficient. Salt-deficient individuals usually lick soil, woods, and even their own urine or that of other individuals [28]. According to the protocol for Punjab urial developed in this study, animals were scored as "mineral-deficient" if they showed any sign of licking objects or urine.
