Abstract The objective of this study was to explore the effects of free-range (FR), part-time free-range (PTFR), and cage system (CS) on behavioral repertoire in Lakha (LK), Mushki (MS), Peshawari (PW), and Sindhi (SN) varieties of Aseel chicken during the growing phase (9 to 18 wk of age). In total, 144 Aseel pullets were allotted to 12 treatment groups in a 3 × 4 (rearing system × Aseel variety) factorial arrangement, according to a randomized complete block design (RCBD). Each treatment group was replicated 3 times with 4 birds in each replicate (12 birds per treatment group). The pullets were randomly marked weekly for identification, and their behavior was observed through the focal animal sampling method. Time spent on different behavioral activities was recorded and converted to a percentage. The data were analyzed using 2-way ANOVA under a factorial arrangement using SAS 9.1, and the behavioral parameters were evaluated. The results indicated greater (P < 0.05) sitting, standing, drinking, preening, and aggressiveness in CS; walking, running, and jumping in PTFR; and foraging and dustbathing in both FR and PTFR, whereas feather pecking was found to be reduced in FR compared with PTFR and CS. Among varieties, PW showed the least feeding/foraging and feather pecking behavior, and greater standing, running, and jumping behavior (P < 0.05). However, SN spent less time in walking and preening, and more time in sitting, drinking, and aggressiveness. Dustbathing was found to be similar in all Aseel varieties (P = 0.135). In conclusion, the PTFR system could be suggested as a substitute for conventional housing systems because it better accommodates normal behavior in Aseel pullets. INTRODUCTION Concern for the welfare of farm animals raised in intensive housing systems is growing (Appleby et al., 2004). Providing more welfare-friendly environments, which may curtail animal suffering, is becoming locally and globally important because of the free trade among different nations (Kim et al., 2017), and Pakistan is no different in facing this transition. Recently, poultry farmers have shifted from conventional to alternative housing systems, such as free-range (FR), enriched colonies, and furnished cages. To ensure the transition to cage-free rearing is smooth, poultry farmers must use innovative strategies for which planning, resources, and logistic techniques are required (Thaxton et al., 2016). Free-range and part-time free-range (PTFR) systems provide natural and vegetation enriched environments to the birds by replacing the cage system (CS), which would improve comfort and facilitate in performing normal behaviors (Altan et al., 2000; Sekeroglu and Sarica, 2010; Diktas et al., 2015). Many global reports, international legislations, and scientific literature suggest that welfare excellence cannot be achieved in poultry maintained in cages or intensive systems (Elson et al., 2012; RSPCA, 2016). Outdoor rearing systems help in minimizing stress in birds and enhance their welfare by providing more opportunities for walking and roaming (Lewis et al., 1997; Fanatico et al., 2006), which will improve their foraging, feed selection, and dustbathing behaviors, and thus their welfare is theoretically enhanced (Ponte et al., 2008). These systems decrease aggressiveness, frustration, and stereotyping in birds when they are given opportunities to forage and dustbathe (Mench et al., 2001). The inability of a chicken to perform dustbathing can be stressful and detrimental to its wellbeing (Fraser and Duncan, 1998). Feather pecking in birds is attributed to several causative factors, and effective strategies are yet to be developed to tackle this problem (Lambton et al., 2010; Wysocki et al., 2010; Lambton et al., 2013). One particular factor exacerbating this vice is the inhibition of foraging activity, such as ground pecking, or lack of environmental stimuli (Dixon et al., 2010; Gilani et al., 2013). The early life experiences of birds in the growing phase are important for the development of feather pecking in the laying phase (Johnsen et al., 1998; Rodenburg et al., 2013). The welfare of chickens is dependent on both the presence of enjoyable activities and the absence of suffering. Aseel chicken varieties are originally from India and are well known for their agility, pugnacity, dogged fighting stamina, and adaptability to the indigenous conditions of the Indo-Pak sub-continent (Singh, 2001). This is a slow-growing breed and considered to be one of the ancestors of Cornish and Plymouth Rock, the parents of today's modern broiler (Dohner, 2001). Being naturally scavengers, Aseel chickens may be used for meat production in FR and PTFR systems. Hens of this breed have an average laying rate of 67.57% at peak production (Haunshi et al., 2013). Cockfighting is illegal in the United States, and Louisiana was the last state to ban this practice in 2007 (www.americancowboychronicles.com/2014/04/its-called-cockfighting.html). Initially, the Aseel chicken was used in cockfighting competitions, but many developing countries, including Pakistan, are now banning this practice (http://www.raising-chickens.org/aseel.html). Among the Aseel varieties existing in Pakistan, Lakha (LK) is the heavier one, characterized by a reddish brown plumage with black or white mottling, with hens varying in BW from 2.5 to 3.2 kg. Mushki (MS) is medium-sized with a black plumage containing a blackish dark pigmentation in beak and shanks, with hens ranging in BW from 2.5 to 3.0 kg. Peshawari (PW) is small in size, having a wheaten-colored plumage with BW fluctuating from 2.2 to 2.5 kg in hens. Sindhi (SN) is the heaviest among all, possessing a reddish brown plumage with hard and short feathers, with hens having BW ranging from 2.75 to 3.5 kg (Babar et al., 2012; Usman et al., 2014). Various behavioral traits, including fearfulness, feather pecking, and aggressiveness, vary from strain to strain or breed to breed (Jones and Hocking, 1999). The welfare of farm animals may be improved through the selection of indigenous breeds adapted to the local inclement environmental conditions (Koknaroglu and Akunal, 2013). Although a lot of work has been conducted on genetics, physiology, and growth rates of the Aseel chicken, there is little information available on its behavior in differing housing systems. Therefore, a pilot study was conducted to compare the behavior patterns of 4 varieties of Aseel chicken (LK, MS, PW, and SN) housed in 3 different rearing systems (FR, PTFR, and CS). We hypothesized that the different varieties of indigenous Aseel chicken may express their behavior differently when maintained in different housing systems. MATERIALS AND METHODS Ethics The experiment was conducted according to the approved protocol and guidelines of the University of Veterinary and Animal Sciences (UVAS) Lahore Animal Ethics Committee, devised for the care and use of experimental animals for scientific purposes. Animals and Housing Two-hundred-sixteen one-day-old female chicks, 54 from each variety, were procured from the hatchery of the Avian Research and Training (ART) Center UVAS Lahore. The early rearing, hatch to 6 wk of age, was performed at the Indigenous Chicken Genetic Resource Center (ICGRC) Ravi Campus UVAS Lahore, in a 4-tiered cage (1.8 m L × 1.2 m W × 0.4 m H, stocking density = 25 birds/m2). The cage was located in an environment-controlled house (temperature ranging from 21 to 32°C and RH ranging from 60 to 75%) and effectively fitted with the adjustable feeders, nipple drinkers, and removable dropping trays. The present study was conducted during the mo of August to October 2015. In total, 144 Aseel pullets, 36 from each variety, were randomly selected and assigned to 12 treatment groups in a 3 × 4 (rearing system × Aseel variety) factorial arrangement under a randomized complete block design (RCBD). Each treatment group was replicated 3 times with 4 pullets per replicate. The experimental animals were properly tagged and maintained in an independent open-sided poultry house (6.11 m L × 6.11 m W, 37.33 m2) oriented north to south, equipped with a 3-tiered rearing cage (4.88 m L × 2.44 m W, 35.72 m2, stocking density = 2.68 pullets/m2). The cage was partitioned into 24 symmetrical units (1.22 m L × 1.22 m W × 0.61 m H) and allocated to PTFR and CS (4 birds per unit). Accessible feeders and nipple drinkers were adequately installed on both sides of the cage. The photoperiod was 12L:12D (0600–1800 h), and it remained constant throughout the experiment. Similar hygienic and prophylactic measures were adopted in all the housing systems. The birds were vaccinated against Newcastle disease virus (LaSota) and infectious bronchitis virus (H 120) according to the locally recommended vaccination schedule. Range Area. A patch of fertile land measuring 15 m L × 34 m W (stocking density = 0.21 pullets/m2) located adjacent to the same open-sided house was used as the range area. Seasonal vegetation, including legumes (beans, lentils, peas, and pulses) and non-legumes (fodders and grasses), was grown in the range for enrichment. The range forage was kept in a growing state by intervallic mowing; otherwise, it would become mature and fibrous. In the ranging area, there were 2 parallel rows of replicates (one for FR and the other for PTFR) oriented north to south, each comprising 12 identical replicate units. The replicates were dimensioned to 15 m L × 1.25 m W × 2.44 m H with a walkway (15 m L × 4 m W) in the middle of both rows. Partitions among replicates of FR and PTFR were made by using fishing nets. Fresh and hygienic water was available ad-libitum through a nipple drinking system installed in all the treatment groups. The birds were protected from the wild animals and predators by installing a 2.44 m high wire-mesh enclosure surrounding the range area. Free-Range, Part-Time Free-Range, and Cage System. The birds reared in the FR system had a “whole-day” access to the range (0800–1600 h) and were shepherded to the same house (stocking density = 1.88 pullets/m2). The FR birds were maintained on a floor (1600–0800 h) with rice husk as bedding material (15 cm) and offered 25% feed allowance (1700 h) of broiler breeder grower ration, formulated according to the recommendations of NRC (1994). The diet was iso-nitrogenous and iso-caloric (CP 17.00%, ME 3,000 kcal/kg), contained 2.61% calcium, 0.93% phosphorus, 1.09% Lys, and 0.45% Meth. The pullets housed in PTFR were given a “half-day” access to the range (0800–1200 h), and subsequently they were maintained in cages (1200–0800 h) with 50% feed allowance (1700 h) in removable trough feeders. An adjustment period of 2 wk (wk 7 to 8) was given to the birds in PTFR, and they were trained by directing them to the range and replacing in cage to avoid subsequent handling stress. Birds in the CS system were maintained 24 h in cage and offered 100% allowance of the same broiler breeder grower ration (0800 h). The average daily indoor and outdoor temperature (27.5°C) did not fluctuate widely and remained in the range of 24 to 31°C and 20 to 35°C, respectively. Natural cover in the range area, such as shady trees, tall forages, seasonal crops, and woody perennials, plays a vital role in the maintenance of the ecosystem, which protects the birds from thermal extremes, stress, and UV exposure (Fanatico et al., 2016). Data Collection In total, 36 Aseel pullets from 3 different housing systems and 4 varieties (12 birds/housing system; 9 birds/variety) were randomly observed every wk (9 to 18 wk of age) during their peak activity time starting early in the morning and ending at noon (0800–1200 h). The behavioral repertoire of the pullets was recorded through the focal animal sampling method (Lehner, 1992) using visual scans. Prior to visual scanning, at least one h elapsed, allowing the pullets to redistribute normally as the observers moved across the replicates. During the observer-training period, the pullets were accustomed to the presence of the observers and the beep sounds of the stopwatch. The pullets were randomly selected (one bird/replicate) and marked on the neck and tip of the tail feathers with a non-toxic white-colored blot for identification. All 3 observers were positioned gently near the pullets, avoiding any interruption in their activities. The birds were observed for a 15-minute duration by using digital stopwatches fixed at 5 s beeps, and the behavioral range was recorded individually (time spent in each behavior). After each observation, the researchers paused for 5 min, allowing the next focal pullet to regain its normal position. The mutually exclusive behaviors coded were: sitting, standing, walking, running, jumping, feeding/foraging, drinking, preening, dustbathing, feather pecking, and aggressiveness (Table 1). A purpose-designed behavior assessment sheet was used to collect information regarding various behaviors of the pullets. Subsequently, the data were incorporated in an MS-excel sheet and presented as a percentage of time spent in different behavioral activities. Table 1. Ethogram of behavioral repertoire in Aseel pullets. Behavior Description1 Sitting Sitting with hocks resting on ground without any extra activity Standing Standing alert, eyes open, without any additional activity Walking Locomotion with normal speed or with quick steps Running An activity of wing-assisted running Jumping Movement of a bird in rebounds by leaping with all feet off the ground Feeding/Foraging A pullet directing its beak into feed trough/toward forages and carrying out pecking, manipulating, or ingesting feed, once or repeatedly Drinking Time spent by a pullet at nipple drinkers, drinking water or manipulating nipple drinkers Preening A pullet directing its beak to the plumage in sequential rotating movements once or repeatedly for straightening feathers through pecking, nibbling, and combing Dustbathing Dustbathing bouts of a pullet either in FR or PTFR, squatting down in the substrate, with the use of wings, head, neck, and legs performing sequential vertical wing shaking Feather pecking Pecking movements of a pullet directed at feathers of its conspecific Aggressiveness A response that delivers somewhat unpleasant, giving or receiving peck forcefully, the beak being above the receiver's head. A pullet chases or is chased by its conspecific in an aggressive manner. Miscellaneous All other extraordinary behaviors not mentioned in the current study Behavior Description1 Sitting Sitting with hocks resting on ground without any extra activity Standing Standing alert, eyes open, without any additional activity Walking Locomotion with normal speed or with quick steps Running An activity of wing-assisted running Jumping Movement of a bird in rebounds by leaping with all feet off the ground Feeding/Foraging A pullet directing its beak into feed trough/toward forages and carrying out pecking, manipulating, or ingesting feed, once or repeatedly Drinking Time spent by a pullet at nipple drinkers, drinking water or manipulating nipple drinkers Preening A pullet directing its beak to the plumage in sequential rotating movements once or repeatedly for straightening feathers through pecking, nibbling, and combing Dustbathing Dustbathing bouts of a pullet either in FR or PTFR, squatting down in the substrate, with the use of wings, head, neck, and legs performing sequential vertical wing shaking Feather pecking Pecking movements of a pullet directed at feathers of its conspecific Aggressiveness A response that delivers somewhat unpleasant, giving or receiving peck forcefully, the beak being above the receiver's head. A pullet chases or is chased by its conspecific in an aggressive manner. Miscellaneous All other extraordinary behaviors not mentioned in the current study 1Modified from Hurnik et al., 1995; Bokkers and Koene, 2003; Zhao et al., 2014. View Large Observer Reliability. For ensuring a high level of inter-observer reliability and synchrony within the observer pairs, prior to the beginning of the study (wk 7 to 8), all 3 researchers were trained together in pairs (comprising 2 of the 3 persons) for conducting direct observations on different behaviors of the pullets. One of the observers was standing inside the open-sided house in front of the cage, and the other 2 were outside in the walkway between FR and PTFR, interchanging them weekly for greater accuracy. Statistics Before analysis, the data were tested for homogeneity, verified for normality, and subjected to the analysis of variance (ANOVA) technique under a factorial arrangement using the GLM procedure (SAS Institute Inc., 2002–03). Housing system and Aseel variety were the main effects, and their interaction was tested for significance. Each replicate was considered as an experimental unit. Comparison among treatment means was made through Duncan's multiple range (DMR) test (SAS Institute Inc., 2002–03) and statistical significance was established at P ≤ 0.05. RESULTS Different rearing systems varied (P < 0.05) in sitting, standing, and walking behaviors (Table 2). The birds housed in CS spent more time in sitting (F2,35 = 313.89, P < 0.0001) and standing conditions (F2,35 = 234.42, P < 0.0001), whereas the birds reared in PTFR spent a greater amount of time in walking (F2,35 = 110.25, P < 0.0001) followed by those housed in FR and CS. The birds reared in PTFR indicated greater time in running (F2,35 = 69.89, P < 0.0001) and jumping (F2,35 = 65.19, P < 0.0001) than those of FR, whereas time spent in feeding was greater (F2,35 = 10.53, P < 0.0023) in both FR and PTFR than CS. The birds maintained in CS spent more time (F2,35 = 10.93, P < 0.0001) at the drinkers than the birds in FR or PTFR. The birds housed in CS demonstrated the highest preening behavior (F2,35 = 136.94, P < 0.0001) followed by those housed in FR and PTFR. The dustbathing behavior could statistically be compared only between FR and PTFR, and the birds showed similar time spent in dustbathing (F2,35 = 1.44, P = 0.2640), both in FR and PTFR. The birds housed in CS and PTFR demonstrated more feather pecking (F2,35 = 12.25, P < 0.0013) than those of FR. Likewise, the birds maintained in CS indicated more aggressiveness (F2,35 = 6.81, P < 0.0106) than those of FR and PTFR. Table 2. Overall behavioral patterns in Aseel pullets housed in 3 different rearing systems (9 to 18 wk of age)1. Behavior4 Effects Housing system2,3 FR PTFR CS P-value Sitting 9.37 ± 0.85b 7.35 ± 0.64c 26.65 ± 0.79a <.0001 Standing 8.02 ± 0.79b 5.97 ± 0.61b 24.62 ± 1.03a <.0001 Walking 15.60 ± 1.99b 24.08 ± 1.71a 0.38 ± 0.06c <.0001 Running 7.71 ± 0.90b 11.41 ± 1.23a 0.00 ± 0c <.0001 Jumping 0.96 ± 0.11b 1.43 ± 0.20a 0.03 ± 0.01c <.0001 Feeding/Foraging 18.47 ± 2.20a 15.57 ± 2.07a 9.62 ± 1.24b 0.0023 Drinking 8.85 ± 1.36b 9.33 ± 1.33b 15.94 ± 1.24a <.0001 Preening 2.81 ± 0.40b 1.33 ± 0.20c 9.38 ± 0.84a <.0001 Dustbathing 13.14 ± 1.24 10.96 ± 1.56 - NS5 Feather pecking 1.04 ± 0.27b 2.09 ± 0.19a 2.13 ± 0.16a 0.0013 Aggressiveness 1.10 ± 0.28b 1.01 ± 0.31b 2.25 ± 0.62a 0.0106 Behavior4 Effects Housing system2,3 FR PTFR CS P-value Sitting 9.37 ± 0.85b 7.35 ± 0.64c 26.65 ± 0.79a <.0001 Standing 8.02 ± 0.79b 5.97 ± 0.61b 24.62 ± 1.03a <.0001 Walking 15.60 ± 1.99b 24.08 ± 1.71a 0.38 ± 0.06c <.0001 Running 7.71 ± 0.90b 11.41 ± 1.23a 0.00 ± 0c <.0001 Jumping 0.96 ± 0.11b 1.43 ± 0.20a 0.03 ± 0.01c <.0001 Feeding/Foraging 18.47 ± 2.20a 15.57 ± 2.07a 9.62 ± 1.24b 0.0023 Drinking 8.85 ± 1.36b 9.33 ± 1.33b 15.94 ± 1.24a <.0001 Preening 2.81 ± 0.40b 1.33 ± 0.20c 9.38 ± 0.84a <.0001 Dustbathing 13.14 ± 1.24 10.96 ± 1.56 - NS5 Feather pecking 1.04 ± 0.27b 2.09 ± 0.19a 2.13 ± 0.16a 0.0013 Aggressiveness 1.10 ± 0.28b 1.01 ± 0.31b 2.25 ± 0.62a 0.0106 a–cMeans with different superscripts in rows within the rearing systems are significantly different (P ≤ 0.05). 1Data have been represented as means ± standard errors. 2FR = Free-range; PTFR = Part-time Free-range; CS = Cage system. 3Data shown represent the treatment means from n = 12 replicates with one pullet in each replicate. 4Behavior of the pullets presented as percentage of time. 5NS = not significantly different (P > 0.05). View Large Among varieties, the SN pullets spent more time in sitting (F3,35 = 6.64, P = 0.0068) compared with those of PW, whereas greater standing time (F3,35 = 4.65, P = 0.0222) was observed in PW than MS, LK, and SN (Table 3). However, the birds of MS, PW, and LK varieties spent more time walking (F3,35 = 4.72, P = 0.0213) than those of SN pullets. The PW variety spent the highest time in running (F3,35 = 3.57, P < 0.0470) as compared with LK and SN. Similarly, higher jumping time (F3,35 = 5.58, P < 0.0124) was observed in PW than LK and SN. Birds of the PW variety spent less time in feeding/foraging (F3,35 = 3.5, P < 0.0495) than SN, MS, and LK, whereas time spent at drinkers was observed to be higher in SN (F2,35 = 3.64, P < 0.0447) than PW. The SN variety spent a greater amount of time in preening (F3,35 = 7.06, P < 0.0054) compared with the others, whereas no difference in dustbathing behavior was observed among the pullets (F3,35 = 2.48, P = 0.1355). Birds of LK, MS, and SN spent more time pecking feathers (F3,35 = 4.87, P < 0.0193) than the PW birds. However, the SN variety spent the most time being aggressive (F3,35 = 10.21, P < 0.0013), followed by LK and PW. Table 3. Overall behavioral patterns in 4 varieties of Aseel pullets (9 to 18 wk of age)1. Behavior4 Effects Aseel variety2,3 LK MS PW SN P-value Sitting 14.86 ± 3.77a,b 13.60 ± 3.69b,c 12.62 ± 4.10c 16.75 ± 4.09a 0.0068 Standing 12.27 ± 3.60b 12.13 ± 3.67b 15.34 ± 4.47a 11.74 ± 3.40b 0.0222 Walking 13.66 ± 5.02a 14.64 ± 4.66a 15.85 ± 5.26a 9.27 ± 3.20b 0.0213 Running 5.32 ± 1.84b 6.87 ± 2.43a,b 8.30 ± 2.98a 5.00 ± 1.69b 0.0470 Jumping 0.60 ± 0.19b 0.89 ± 0.32a,b 1.11 ± 0.39a 0.63 ± 0.21b 0.0124 Feeding/Foraging 15.40 ± 2.37a 16.30 ± 2.02a 10.10 ± 1.15b 16.43 ± 3.77a 0.0495 Drinking 11.76 ± 1.99a,b 12.12 ± 1.41a,b 7.68 ± 1.50b 13.93 ± 2.38a 0.0447 Preening 4.90 ± 1.80a 4.92 ± 1.80a 5.36 ± 1.79a 2.85 ± 1.00b 0.0054 Dustbathing 12.49 ± 1.71 9.07 ± 2.13 15.78 ± 1.32 10.87 ± 1.64 NS5 Feather pecking 2.09 ± 0.37a 1.89 ± 0.28a 1.09 ± 0.26b 1.94 ± 0.22a 0.0193 Aggressiveness 1.49 ± 0.43b 1.04 ± 0.20b,c 0.50 ± 0.12c 2.79 ± 0.70a 0.0013 Behavior4 Effects Aseel variety2,3 LK MS PW SN P-value Sitting 14.86 ± 3.77a,b 13.60 ± 3.69b,c 12.62 ± 4.10c 16.75 ± 4.09a 0.0068 Standing 12.27 ± 3.60b 12.13 ± 3.67b 15.34 ± 4.47a 11.74 ± 3.40b 0.0222 Walking 13.66 ± 5.02a 14.64 ± 4.66a 15.85 ± 5.26a 9.27 ± 3.20b 0.0213 Running 5.32 ± 1.84b 6.87 ± 2.43a,b 8.30 ± 2.98a 5.00 ± 1.69b 0.0470 Jumping 0.60 ± 0.19b 0.89 ± 0.32a,b 1.11 ± 0.39a 0.63 ± 0.21b 0.0124 Feeding/Foraging 15.40 ± 2.37a 16.30 ± 2.02a 10.10 ± 1.15b 16.43 ± 3.77a 0.0495 Drinking 11.76 ± 1.99a,b 12.12 ± 1.41a,b 7.68 ± 1.50b 13.93 ± 2.38a 0.0447 Preening 4.90 ± 1.80a 4.92 ± 1.80a 5.36 ± 1.79a 2.85 ± 1.00b 0.0054 Dustbathing 12.49 ± 1.71 9.07 ± 2.13 15.78 ± 1.32 10.87 ± 1.64 NS5 Feather pecking 2.09 ± 0.37a 1.89 ± 0.28a 1.09 ± 0.26b 1.94 ± 0.22a 0.0193 Aggressiveness 1.49 ± 0.43b 1.04 ± 0.20b,c 0.50 ± 0.12c 2.79 ± 0.70a 0.0013 a–cMeans with different superscripts in rows within the varieties are significantly different (P ≤ 0.05). 1Data have been represented as means ± standard errors. 2LK = Lakha, MS = Mushki, PW = Peshawari, SN = Sindhi. 3Data shown represent the treatment means from n = 9 replicate with one pullet in each replicate. 4Same as in Table 2. 5NS = not significantly different (P > 0.05). View Large However, the interaction of housing system and Aseel variety showed similarities (P > 0.05) in sitting (F6,35 = 0.40, P = 0.8676), standing (F6,35 = 1.11, P = 0.4095), walking (F6,35 = 1.82, P = 0.1784), running (F6,35 = 1.81, P = 0.1795), feeding/foraging (F6,35 = 2.67, P = 0.0694), drinking (F6,35 = 0.40, P = 0.8655), preening (F6,35 = 2.27, P = 0.1067), dustbathing (F6,35 = 0.50, P = 0.6953), feather pecking (F6,35 = 0.61, P = 0.7193), and aggressive (F6,35 = 2.26, P = 0.1075) behaviors except for jumping, which varied significantly (F6,35 = 3.45, P = 0.0323), showing a greater fraction of time in PW and MS housed in PTFR. DISCUSSION How the pullets are reared affects welfare throughout their remaining lives. A rearing system that provides hens with better welfare, for instance, cage-free/FR delivers the birds with a greater potential to express natural behaviors (Janczak and Riber, 2015). In the FR system, birds have an ample space to express their normal behaviors, whereas the CS offers little opportunity for birds to manifest their full behavioral repertoire, particularly running, jumping, scratching, and dustbathing. Effect of Housing System on Pullet Behavior In the present study, increased sitting and standing behaviors in CS might be linked to the higher stocking density, leading to impaired activities. A previously conducted study (Irfan et al., 2016) reported enhanced sitting and standing behaviors in turkeys (Maleagris gallopavo) reared in confinement compared with the FR. The higher walking activity observed in the PTFR system was supported by the findings that the range environment helps in reducing stress and improving comfort in birds by providing them complete freedom for walking or roaming (Fanatico et al., 2006; Kperegbeyi et al., 2009) together with leg stretching, perching, and wing flapping, which enhanced their locomotion (Mench et al., 2001). Similarly, Irfan et al. (2016) reported more immobility in birds maintained in confinement compared with those of FR. Previously, there have been studies on different species of birds showing the ameliorative effects of lower stocking densities per se (Sanotra et al., 2002; Buijs et al., 2009: Marchewka et al., 2013). Apparently, at higher stocking densities, birds are less active due to more intervention in their locomotion. Likewise, in the present study, discrepancies in locomotion might have come about just owing to the lower stocking densities in both FR and PTFR (0.21 pullets/m2) compared with CS (2.68 pullets/m2). More jumping and running activities observed in the birds housed in PTFR might be attributed to their intrinsic drive for a capacious environment. It was speculated that the birds in PTFR on having access to the range might have shown their natural frolicking activity (Jones and Mills, 1999; Marin et al., 2001; Goth and Jones, 2003). Similarly, higher locomotion was reported in slow-growing chickens maintained in outdoor production systems (Dal Bosco et al., 2012). An enhanced feeding/foraging behavior was observed in the pullets housed in FR and PTFR, which might be attributed to the diversity of stimuli in the range, promoting foraging activity in birds (Knierim, 2006). The foraging behavior experienced by the pullets at an early age might be helpful in minimizing the risk of feather pecking in their laying stage (Gilani et al. 2013). Moreover, the FR systems, due to the availability of an ample space (Lewis et al., 1997; Fanatico et al., 2006) provide more opportunities for curiosity, exploration, and foraging to the birds than the single-tiered aviary (Shimmura et al., 2008). Corroborating these findings, greater foraging activity in turkeys (Irfan et al., 2016) and promoted ranging behavior in broilers were observed in FR compared with confinement (Ponte et al., 2008). Similarly, the pullets spent more time at drinkers in CS, which may be correlated to the availability of concentrated rations increasing the birds’ thirst, leading them to drink more frequently (Irfan et al., 2016). More preening behavior observed in the pullets housed in CS might be speculated due to the higher stocking density in cage. Preening in the cage is sometimes perceived as a displacement behavior due either to the situation of conflict or environmental frustration (Duncan and Wood-Gush, 1972), which corresponds to the anecdotal avian knowledge. Likewise, Shimmura et al. (2008) reported more proportions of hens performing preening in a single-tiered aviary (SA) compared with the FR hens, indicating that preening and intensive systems are positively correlated (Elston et al., 2000). Dustbathing is a comfort behavior, characterized by the act of grooming when the pullets move around in sand or dust, facilitating them in maintaining body temperature, insulating, and helping in controlling ectoparasites by absorbing excess oil and then shaking it off. Domestic chickens (Gallus domesticus) contain intrinsic motivation to dustbathe for cleaning their feathers (RSPCA, 2016). In the current study, dustbathing was observed unambiguously in FR and PTFR. CS are deemed unsuitable for the birds, as they allow only for poor expression of natural behaviors, including dustbathing (Appleby et al., 2004). Feather pecking in pullets is a non-aggressive behavior through which the birds peck at the feathers of their conspecifics (Hartcher et al., 2015). Lack of salient external stimuli (e.g., litter, dust, sand, food particles) for dustbathing or inhibition of foraging activity, including ground pecking, is considered an essential factor associated with the feather pecking (Vestergaard et al., 1993; Dixon et al., 2010; Gilani et al., 2013). Less feather pecking observed in FR was apparently due to the availability of extensive space for exercise allowing birds to express their natural behaviors, which might reduce chances of feather pecking. Similar to these findings, Huber-Eicher and Sebo (2001) observed a minimum incidence of feather pecking in birds when roaming and foraging. However, in battery cages, the birds are unable to exercise, peck the ground/forage, ultimately leading to more feather-pecking behavior (Rodenburg et al., 2013). In our study, more aggressiveness was observed in the pullets maintained in CS. The plausible reason might be that the pullets could not exhibit a complete suite of behavioral activities due to scarcity of space, resulting in more antagonistic behavior. Duncan and Wood-Gush (1972) reported that a situation of conflict or frustration in light hybrid strains of hens maintained in cages might lead to stereotyped pacing and increased aggressiveness. On the other hand, FR housing systems decrease frustration and aggression, which might be correlated to the enhanced dustbathing and ground-scratching activities in birds (Mench et al., 2001). Effect of Breed on Pullet Behavior Selecting a suitable breed for the outdoor systems is a necessary complement of an efficient management strategy. Slow-growing robust chicken breeds might be a good choice for the FR systems when the birds are kept in small flocks. Our results on sitting and standing behavior coincide with another study that corresponds to discrepancies in these behaviors among different genotypes of indigenous chicken (Ha et al., 2011), demonstrating that behavioral patterns in birds are genotype dependent (Craig and Muir, 1998; Jones and Hocking, 1999). Relatively more standing noticed in PW might be attributed to its comparatively lower BW, which might be the plausible reason behind its superior standing ability and increased stamina. Slow-growing breeds are well suited to the range environment. In our study, the greater foraging behavior observed in heavier varieties (LK, MS, and SN) coincides with the findings of many other scientists who reported more inclination of the slow-growing chickens to enhanced foraging activity in FR (Nielsen et al., 2003; Jones et al., 2007; Almeida et al., 2012). A study conducted on behavior of the slow-growing genotypes with different growth rates indicated that the slow-growing birds were more active, interested in the observer, and made use of the pasture more efficiently (Lewis et al., 1997; Dal Bosco et al., 2012). Walking behavior varied among different Aseel varieties, being the highest in PW, which might be attributed to the distinct variations in their genetic makeup. Likewise, different breeds of poultry also were reported to show an unambiguous disparity in their walking behavior (Dal Bosco et al., 2012). Among varieties, greater running time (P < 0.05) was revealed by PW, which might be attributed to its comparatively lighter BW (personal observation). Disparities in running behavior of different breeds or strains of poultry also were reported by Craig and Muir (1998) and Jones and Hocking (1999). Comparatively, the highest fraction of time spent in jumping was observed in PW and MS, and the plausible reason behind their enhanced jumping activity might be the lower BW. Additionally, the genetic makeup of PW and MS is considered a potential cause of their greater jumping activity, as reported by geneticists earlier that the behavioral patterns in birds are genotype dependent (Craig and Muir, 1998; Jones and Hocking, 1999). The PW variety revealed less feeding/foraging activity to fulfill its nutrient requirements in shorter intervals of time, which was corroborated by a previously conducted study on feeding behavior in different genotypes of poultry (Dal Bosco et al., 2012). PW spent less time at drinkers, which might possibly be correlated to its lower feed intake depending on its body requirements (personal observation). Likewise, different genotypes of poultry have already been reported to show disparity in their drinking behavior (Ha et al., 2011). In our study, less preening in SN and feather pecking in PW were observed, which might be linked to their distinct genetic makeup, as reported previously that preening varied from strain to strain (Craig and Muir, 1998; Jones and Hocking, 1999) and genotype to genotype (Ha et al., 2011). Feather pecking is reported as a heritable trait in poultry (Bessei and Kjaer, 2015). It was earlier reported that quality husbandry practices paired with genetic selection might help in reducing feather pecking (Rodenburg et al., 2013; Bessei and Kjaer, 2015). The more aggressiveness observed in SN might be associated with its genetic predisposition, as reported earlier that some genotypes (Ha et al., 2011) or strains of poultry were more aggressive and thwarting than others (Craig and Muir, 1998; Jones and Hocking, 1999). Our findings on aggressive behavior in Aseel pullets contrasted with those of Ball and Balthazart (2008) who reported that aggressive behaviors among birds are independent of BW. The non-genetic factors affecting normal behavior in pullets may include time of d, season, environmental temperature, weather conditions, and flock size (Almeida et al., 2012). In all, our findings suggest that PTFR is an effective alternative system and raising Aseel pullets in PTFR could be recommended to substitute the currently used conventional housing systems. This novel approach might help in ameliorating normal behaviors, mitigating stress, and combating production-related problems in Aseel pullets during their upcoming laying cycles. ACKNOWLEDGEMENTS The authors gratefully acknowledge the cooperation and support extended by Indigenous Chicken Genetic Resource Center (ICGRC) Department of Poultry Production Ravi Campus UVAS Lahore, and Higher Education Commission (HEC) Government of Pakistan, Islamabad for facilitating and funding the present research. REFERENCES Almeida G. F., Hinrichsen L. K., Horstead K., Thamsborg S. M., Hermansen J. E.. 2012. 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