Spatial Distribution and Dietary Niche Breadth of Leopard Cats (Prionailurus bengalensis) Inhabiting Margalla Hills National Park, Pakistan

© 2021 Academia Sinica, Taiwan Open Access Spatial Distribution and Dietary Niche Breadth of Leopard Cats (Prionailurus bengalensis) Inhabiting Margalla Hills National Park, Pakistan Hira Fatima1, Tariq Mahmood1,* , Lauren Mae Hennelly2, Muhammad Farooq1, Faraz Akrim3, and Muhammad Sajid Nadeem4 1Department of Wildlife Management, PMAS Arid Agriculture University Rawalpindi 43600, Pakistan. *Correspondence: E-mail: [email protected] (Mahmood). Tel: +923215105689. E-mail: [email protected] (Fatima); [email protected] (Farooq) 2Mammalian Ecology and Conservation Unit, University of California, Davis, USA. E-mail: [email protected] (Hennelly) 3Zoology Department, University of Kotli, Kotli, Azad Jammu & Kashmir, Pakistan. E-mail: [email protected] (Akrim) 4Department of Zoology, PMAS Arid Agriculture University Rawalpindi 43600, Pakistan. E-mail: [email protected] (Nadeem) Received 21 December 2020 / Accepted 1 April 2021 / Published 5 July 2021 Communicated by Jian-Nan Liu The leopard cat (Prionailurus bengalensis) is distributed throughout the Himalayan foothills in Pakistan and occurs in moist temperate and dry coniferous forests. However, the cat species is categorized as “Data Deficient” in Pakistan. In the current study we aimed to investigate the leopard cat distribution and dietary niche in the Margalla Hills National Park, the lowest part of the Himalayan foothills in Pakistan. We recorded direct (field observations, camera trapping) and indirect signs (scats) of the species on 23 trails / tracks in the study area. The leopard cat was recorded at 13 different sampling sites in the park, with an altitudinal range between 664–1441 m asl. The diet composition of the species was investigated through scat analysis, with the species identity of the scats confirmed through the mitochondrial cytB region. The species’ diet comprised both animal and plant matter. The animal-based diet (in terms of frequency of occurrence) contained wild (51.75%) as well as domestic prey (7.69%), and plant species (31.47%). Wild prey included small mammals (rodents, two mongoose species, Asian palm squirrel, and Cape hare), birds, insects and snails. The domestic prey were poultry birds, sheep, goats and dogs. Consumption of wild prey was higher in summer (n = 31 scat), whereas intake of domestic prey was higher in winter (n = 37 scats). The dietary niche breadth was wider 14.84 in summer than winter 10.67. A chi-square test showed a significant difference in seasonal dietary intake of the leopard cat. The study concludes that the leopard cat feeds mainly on wild animal prey and plant species; however, in winter consumption of domestic prey increases. Key words: Spatial distribution, Seasonal diet, Dietary composition, Prey species, Niche breadth. BACKGROUND There are two leopard cat species recognized by the IUCN Cat Specialist Group: the mainland leopard cat (Prionailurus bengalensis) and the Sunda leopard cat (Prionailurus javanensis). The mainland leopard cat is a relatively common and widespread small wild cat (Sunquist and Sunquist 2002; Ross et al. 2015) that is distributed throughout the South and East Asian mainland (Nowell and Jackson 1996). The leopard cat (Prionailurus bengalensis) is a relatively common and widespread small felid (Sunquist and Sunquist 2002) distributed throughout Southeast Asia (Nowell and Jackson 1996). However, its population status varies Citation: Fatima H, Mahmood T, Hennelly LM, Farooq M, Akrim F, Nadeem MS. 2021. Spatial distribution and dietary niche breadth of leopard cats (Prionailurus bengalensis) inhabiting Margalla Hills National Park, Pakistan. Zool Stud 60:34. doi:10.6620/ZS.2021.60-34. Zoological Studies 60:34 (2021) doi:10.6620/ZS.2021.60-34 1 © 2021 Academia Sinica, Taiwan across the countries in its range, being endangered and critically endangered in Korea and Japan, respectively, comparatively stable in China and some parts of India, and data deficient in the Himalayan foothills Afghanistan and Pakistan (Shehzad et al. 2012; Ross et al. 2015). In Pakistan, the leopard cat is mostly associated with Himalayan moist temperate forest, but also dwells further north in the Himalayan dry coniferous forest (Roberts 1997). Because no recent data are available on its population in Pakistan, the status of this species is categorized as “Data Deficient” (Sheikh and Molur 2005). The main threats to this species in Pakistan include hunting by commercial traders for its richly spotted fur mainly for decorations and coats, captive breeding as pets, interbreeding with domestic cats to make the domesticated Bengal cat and traditional medicinal use of bones (Nowell and Jackson 1996). More recently, this species continues to be hunted throughout most of its range for fur, for food and domestication (Ross et al. 2015). The species also faces retaliatory killing for hunting poulty. Island populations are small and seriously threatened in the Philippines and Japan. The leopard cat consumes a broad spectrum of prey, hence its diet generally constitutes a variety of prey species including small mammals such as shrews and squirrels, birds, reptiles and fish, reflecting the cat’s prey flexibility and habitat diversity in its diet (Grassman et al. 2005; Shehzad et al. 2012). Although the overall diet of the leopard cat is diverse and flexible, there are regional differences across the leopard cat range. In Sabah and Borneo part of Malaysia, the main prey of the leopard cat are mammals (murids), with Whitehead’s rat (Maxomys whiteheadi) being the principal prey species. The preference for Whitehead’s rat was also suggested to relate to its ‘catchability’ rather than its high prey density in oil palm plantations (Rajaratnam et al. 2007). Similarly, in the forests of Singapore, the leopard cat diet mainly consists of mammalian prey, with Oriental house rat (Rattus tanezumi) found in all scats (Chua et al. 2016). Insects were the second most frequently consumed food group (52.3% FOS), among which orthoptera appeared in 29.2% of scats, reptiles (18.5%), birds (15.38%) and amphibians (1.54%) were ranked third through fifth in terms of FOS. Although the leopard cat is generally flexible in its habitat choice and prey selectivity it still faces many threats including habitat loss due to anthropogenic activities, commercial exploitation for fur trade and competition with sympatric species (Izawa 1991). Khan (2009) reported a leopard cat occurrence in Margalla Hills National Park Islamabad, Pakistan along with other 13 mammalian species. However, the park lacks a comprehensive study on leopard cat distribution and diet and there is paucity of information on how food selection is made by this felid species. Therefore, the current study aimed to confirm the occurrence of leopard cat in Margalla Hills National Park, Islamabad, and analyze its diet composition, and niche breadth, through scat analysis. MATERIALS AND METHODS Study area The current study was conducted in the Margalla Hills National Park (MHNP) Islamabad, Pakistan, situated along the northern border of Federal Capital city, Islamabad at 33°43'N longitude and 73°55'E latitude (Fig. 1). Area of the park is approximately 17,386 ha (UNDP/IUCN/MINFA/CDA, 1991), having rugged topography comprising mainly of steep slopes and gullies with elevation ranging from 450 to 1,580 m asl (Jabeen et al. 2009). Its climate is an atypical version of a humid subtropical, with hot summers followed by monsoon season, and accompanied by mild and wet winters. It experiences two rainy seasons each year; summer monsoon (July–September) with heavy rainfall and evening thunderstorms and winter rain (January– March). Average minimum and maximum temperatures are 19.5°C and 33.3°C, respectively (Hussain 1986) and mean annual rainfall is about 94 cm. Average relative humidity during the monsoon period varies between 59% and 67% (Masroor 2011). Study design The occurrence of the leopard cat in the park was recorded by using the “sign survey” method, for which both direct (direct field observations, camera trapping) and indirect signs (mainly scats) of the species were recorded during field surveys following Wemmer et al. (1996). A total of 23 trails / tracks in the park were walked on foot to record the species signs from September 2015–December 2018 (Fig. 1 A, B). We also performed a camera trap survey for 3 months, for which we installed a total of seven (07) camera trap stations (Bushnell Trophy Cam HD Essential 2) in the study area along the selected trails with the most leopard cat activity to collect photographic evidence of its occurrence in the study area. During field visits, information on direct sightings, road kills, camera traps as well as fecal samples (data on site, geographic location, elevation, date and species identification for each scat) of the leopard cat were collected to map the spatial distribution of the species using Quantum page 2 of 12Zoological Studies 60:34 (2021) © 2021 Academia Sinica, Taiwan Geographical Information System (QGIS: Version 2.18) software. During field surveys, we collected scats of the leopard cat wherever we encountered it; these were later analyzed to identify leopard cat species through genetic analysis and to investigate the dietary habits of the species. Scat collection To obtain a detailed account of the leopard cat diet, its scat samples were collected year-round through regular surveys on specified trails (23 of selected trails and transects) from September 2015–December 2018. Each field survey was conducted by a team of three to four members including the author and field staff of Islamabad Wildlife Management Board (IWMB) to correctly identify scat morphology and other activity signs of carnivores. Initially, encountered scat morphology was identified in the field, relying on its smell, shape, length, diameter and other physical appearance (such as color and contents of scat e.g., hairs, bones) following Jackson and Hunter (1996). Additionally, deposition site of the scat sample (either on rock or leaves) and other activity signs (scrapes, rubbings and footprints) were also taken into account. All scat samples were collected in self-sealing plastic bags and selected samples were separated and preserved in 95% ethanol for molecular analysis (which were fresh and intact). After the cats were dried, their morphological characteristics such as length, breadth and weight were recorded in the laboratory of the Department of Wildlife Management, PMAS-AAUR for initial investigation and storage. Fig. 1. (A) Map of Pakistan, illustrating the study area, the Margalla Hills National Park (MHNP), Islamabad. (B) Enlarged map of study area, illustrating the 23 transects and trails surveyed during the study period. (C) Map showing spatial distribution of the leopard cat in Margalla Hills National Park Islamabad, Pakistan Islamabad during the present study. N page 3 of 12Zoological Studies 60:34 (2021) © 2021 Academia Sinica, Taiwan Molecular identification of scats Molecular identification of leopard cat scats was carried out in the “Mammalian Ecology and Conservation Unit (MECU)” of the Veterinary Genetics Laboratory at University of California, Davis, USA, for species identification by sequencing ~425 bp of the mitochondrial cytochrome b region. For DNA extraction, we used QIAamp DNA Stool Mini Kits (Qiagen, INC., Valencia, CA). To keep track of possible contamination for each extraction set (12–18 samples) we included a negative control. We used the primer combination of RF14724 and RF15149 (Perrine et al. 2007) to produce 425-bp-long sequences of cytochrome B (5′ portion), which became 354 bp after trimming. The PCR thermal cycler program conditions were denaturation at 94°C for 10 min., then 40 cycles of PCR starting at 94°C for 30 sec., then annealing at 50°C for 30 sec. and elongation at 72°C for 45 sec. These cycles were followed by a 10-min extension step at 72°C. PCR product was cleaned up by performing ExoSAP reaction adding exonuclease I (Exo) and recombinant shrimp alkaline phosphatase (rSAP). We mixed ExoSAP in a micro centrifuge tube and in an appropriately labeled PCR plate. 1 µl od ExoSAP was added to each well while it was placed on ice beads. After a brief spin, we added PCR product to corresponding wells in the ExoSAP plate. Carefully, we sealed the plate and ran exo/sap Program: 37°C for 30 minutes, 80°C for 15 minutes, 15°C forever (ExoSAP recipe for 96 samples). We sequenced the PCR product in the forward direction using Big Dye Terminator v3.1 (Applied Biosystems, Foster City, CA). We placed an ABI plate containing the sequencing product in an ABI 3730 capillary sequencer (Applied Biosystems, Foster City, CA). We used Sequencher 5.4 to import all files from the AB1 folder as well as necessary cytb files (of the leopard cat) to read the sequences file for each sample. Once we got the sequences from all samples, we used Basic Local Alignment Search Tool (BLAST) in GenBank to identify the leopard cat species. The field accuracy rate of the species was calculated by using the formula: sum of true positives and true negatives / the sum of all possible outcomes. Scat analysis Faecal samples of the leopard cat were analyzed in the lab using protocol following Kelly (1991). Each scat was emptied from its plastic bag into a 2.5 mm sieve positioned over a metal tray. Faecal residue was separated by hand for bone, hair, teeth, toenails, feather and diagnostic plant parts (Moore et al. 1974). The mass of each dietary item including hairs, bones, feathers, insects and plant parts were recorded using an electronic weighing balance to compute percent volume. The weights of different remains (e.g., bones, hairs, seeds) in fecal samples reflect the intake that was not digested, and actually not the amount digested. However, it is useful to quantify the left-over undigested remains of food to use these measures to estimate the frequency and volume of different food components ingested. Therefore, although it is not appropriate to use weight as a proxy for intake proportions, weight does reflect the food item consumed by the animal, and the weights of remains measured can be useful in calculating the food items preferred and most or least consumed. Thus, it is assumed that the food remain that are represented maximum in scats, is most heavily consumed and preferred one. However, this method has important limitations because any food item that is totally consumed may not be reflected in the scat remain. Prey species of the leopard cat were identified using medullary pattern of the hairs recovered from scat samples as described by Moore et al. (1974), and a scale replication following Lavoie (1971). Hairs of mammalian prey were identified using light microscope. These prepared slides of samples hairs were then compared with reference hair slides for identification. These reference slides are a collection of light microscopic slides hairs of known mammalian species maintained in the “Mammalian Ecology and Conservation Lab” of the Department of Wildlife Management, PMAS-AAUR, Pakistan. Similarly, other segregated parts (such as bones, feathers, insects) were also compared to reference material for correct identification. Plant remnants recovered from scats were comprised mainly of fruit and seeds eaten by leopard cat, and all recovered seeds and fruit residues along with reference material (known plant material collected from the study area) for identification were sent to Department of Plant Sciences, Quaid-eAzam University, Islamabad in Plant Systematic and Biodiversity Lab for correct identification of plant remains. Dietary niche breadth We computed the dietary niche breadth of the leopard cat species using standardized Levins index (Lst) (Colwell and Futuyma 1971; Levins 1968) based on the following formulae: Σ n i = 1 -1 pi 2 L = ( ) and L−1/n−1 page 4 of 12Zoological Studies 60:34 (2021) © 2021 Academia Sinica, Taiwan Prey species indices Similarly, the prey indices including prey species richness (S), diversity index (H') and prey evenness (E) indices were computed using the following formulae: Diversity Index (H') H'= -Σ [pi × ln pi] where pi represents the prey index and ln pi is the natural log of pi. Prey species richness was calculated by considering the total number of prey species consumed by each carnivore in a specific season. Prey Evenness Index was calculated by using the formula: E = H' / ln of S where, S represents the prey species richness and H' represents the diversity index. Statistical analysis We used the Chi-square test for independence, also called Pearson’s chi-square test, to assess if there is any statistical relationship between two categorical variables. Chi square analysis was performed to assess if there were significant differences (p < 0.05) in the consumption of wild prey species, domestic animals, plant matter and anthropogenic matter in the diet of leopard cat species between summer and winter. We categorized the leopard cat diet into four categories: wild prey, domestic prey, plant materials, and anthropogenic matter; using the total frequencies of each dietary category, we compared dietary intake season wise, that is, in summer and winter, to check if dietary intake differed seasonally. All analysis was performed in SPSS software (Version 20). RESULTS Spatial distribution Among all surveyed sampling sites (N = 23), leopard cat (Prionailurus bengalensis) occurrence was recorded at 13 different sites; evidence consisted of scats, direct sightings, camera traps, and carcasses (Table 1; Fig. 1C; Fig. 2). The altitudinal range of leopard cat distribution in the MHNP was between 664–1441 m. Scats of the species (N = 68) were found at all 13 positive sampling sites. The leopard cat was also sighted in the field at three sampling sites while one carcass was recovered from Trail-5A in the study area. The highest number of field signs of leopard cat was also recorded at Trail-5A. The leopard cat species was camera trapped at only one site, and that camera trap site is shown as blue “asterisk” (*) in figure 1C; one leopard cat carcass was found at one site (Trail-5), shown as a red triangle, while all rest of the sites contained leopard cat scats (Fig. 1B and C). Diet composition Physical characteristics of scats The collected leopard cat scat scats (N = 68) were measured in the laboratory; length = 9.76 ± 0.05 cm, breadth = 1.21 ± 0.01 cm and weight = 7.36 ± 0.04 g. Molecular identification of scats Out of the 68 total scats collected from the field, 56 were confirmed to belong to the leopard cat based on the mitochondrial cytochrome B region. Remaining n = 12 scats were not subjected to molecular analysis just to reduce the cost of analysis, since those scats were collected from the same latrine of the leopard cat and confirmed in the field morphologically based on their shape, size and contour. Similarly, those scats that were found together (12) were also not processed for genetic analysis. However, these 12 scats were included in the final diet analysis of the species. Table 1. Signs of the leopard cat recorded at Margalla Hills National Park Islamabad, Pakistan Type of evidence Numbers recorded (N) Presence recorded (Trails) 13 Scats (N) 68 Direct field sightings 3 Camera trap success 1 Road kills / carcass 1 page 5 of 12Zoological Studies 60:34 (2021) © 2021 Academia Sinica, Taiwan Diet group Analysis of leopard cat scats (N = 68) in the laboratory revealed that they comprised (in terms of percent volume occurrence in scats) of bones, hairs, plant remains, feathers, insects body parts, snails, and anthropogenic matter (Table 2). Bones and hairs were the most voluminous items recovered from the scats analyzed, followed by plant remains. The scats primarily contained remnants of rodents, and a variety of plant species (Table 3). The leopard cat consumed several species of rodents, including house rat (Rattus rattus), house mouse (Mus musculus), Indian gerbil (Tatera indica), Indian Musk Shrew (Suncus murinus), and Indian Mole Rat (Bandicota bengalensis) along with Asian palm squirrel (Funambulus Fig. 2. (A) Camera trapped photograph of leopard cat in MHNP. (B) a carcass of leopard cat recovered from the study area. page 6 of 12Zoological Studies 60:34 (2021) © 2021 Academia Sinica, Taiwan palmarum). Rodents comprised 67.70% (FO) of the diet, more in summer (87.09%) than in winter (51.35%). Wild carnivora in the leopard cat diet were represented up to 10.29%, lagomorphs 4.41%, wild ungulate (wild boar) 2.94%). Invertebrates contributed 13.23% to the leopard cat diet including snails and insects. The diet is comprised of wild prey (including 10 mammal species, birds, insects, and snails), domestic prey (4 species), plants (13 species), and anthropogenic matter (plastic bag material mostly). Wild prey was most frequently consumed (51.75%), followed by plant material (32.41%) that could have been secondarily ingested by eating prey, and domestic prey species (7.69%) (Fig. 3). Among wild prey species, house rat (20.59%) was most frequently eaten, followed by house mouse (11.76) and Indian musk shrew (10.26%). The other wild prey species eaten were small Indian mongoose, grey mongoose, Asian palm squirrel and Cape hare. The wild boar was most probably scavenged by the leopard cat. Birds, insects and snails also contributed in the diet of leopard cat. Among domestic prey items, poultry consumption was highest (5.88%), followed by sheep, goat and dog. Among 13 plant species consumed, most frequently utilized was the daisy (Bellis perennis; 30.88%). The share of anthropogenic matter in the leopard cat diet was approximately 12% (Table 2). Seasonal variation in diet Among the 68 scat samples of leopard cat, 31 were collected in summer while 37 were collected in winter. In general, consumption of wild prey was higher in summer than winter. Similarly, intake of domestic prey was higher in winter than summer. Plant matter intake was higher in winter (97.30%) than summer (35.48%). Prey species richness for the leopard cat was comparatively higher in winter but lower in summer. Similarly, the diversity index was also slightly higher in winter than summer. The prey species evenness index was higher in summer than winter (Fig. 4). Chi-square test for independence showed a significant difference (χ2 = 20.304, d.f. = 3, p = 0.0001; α = 0.05) in dietary intake between summer and winter seasons. Niche Breadth Leopard cat’s dietary niche breadth (L) and standardized niche breadth (Lst Values 0-1) were found wider (L = 14.84, and Lst = 0.43) during summer season but narrower during the winter (L = 10.67, and Lst = 0.30) while the total niche breadth of the leopard cat was L = 17.52 and Lst = 0.52. DISCUSSION In the current study, we recorded different signs of occurrence of the leopard cat at 13 different sampling sites (trails) in the study area, including field sightings (n = 3), camera trap photographs (n = 1), road kills (n = 1), and scats (n = 68). Most leopard cat occurrence data were recovered around dense vegetation areas that were away from the villages’ outskirts, reaffirming previous findings that the leopard cat prefers areas of thick vegetation cover (Roberts 1997). Based from our signs, the species does not occur uniformly in the MHNP, but its population was found confined to some areas associated with dense and thick forests across the elevational range of 664–1441 m. In Pakistan, the leopard cat is normally associated with Himalayan moist temperate forests, but it also penetrated the Himalayan dry coniferous forests and into dry sub-tropical scrub forests in the foothills (Roberts 1997). Margalla Hills National Park, in this context, comprises of the foothills of the front range of the Himalayas; therefore, MHNP may be the edge of leopard cat distribution in Pakistan. Table 2. Food items (% volume) recovered from leopard cat scats in Margalla Hills National park, Islamabad, Pakistan Recovered Prey items S (n = 31) W (n = 37) Mean ± SE Bones 45.61 34.72 39.69 ± 0.02 Hairs 28.23 21.36 24.49 ± 0.02 Feathers 3.42 6.49 5.09 ± 0.02 Insects Parts 1.30 3.77 2.64 ± 0.01 Plant Remains 11.90 25.86 19.50 ± 0.02 Soil/ Sand 4.69 0.00 2.14 ± 0.01 Snails 0.00 5.35 2.91 ± 0.01 Anthropogenic matter 5.40 2.07 3.59 ± 0.01 *S= Summer season; *W= Winter season. page 7 of 12Zoological Studies 60:34 (2021) © 2021 Academia Sinica, Taiwan Table 3. Frequency of occurrence (%) of prey items identified from the scats of the leopard cat Prey Species S (n = 31) W (n = 37) (%) F Wild Prey Rodents House Rat (Rattus rattus) 29.03 13.51 20.6 House mouse (Mus musculus) 16.13 8.11 11.8 Indian Gerbil (Tatera indica) 12.90 8.11 10.3 Indian Musk Shrew (Suncus murinus) 12.90 8.11 10.3 Indian Mole Rat (Bandicota bengalensis) 9.68 2.70 5.88 Palm squirrel (Funambulus pennantii) 6.45 10.81 8.82 Sub-Total 87.09 51.35 67.70 Carnivora Small Indian mongoose (Urva auropunctatus) 9.68 5.41 7.35 Grey mongoose (Herpestes edwardsii) 3.23 2.70 2.94 Sub-Total 12.91 8.11 10.29 Lagomorpha Cape hare (Lepus capensis) - 8.11 4.41 Sub-Total - 8.11 4.41 Wild ungulates Wild boar (Sus scrofa) 6.45 - 2.94 Sub-Total 6.45 - 2.94 Invertebrates Snails - 8.11 4.41 Insects 6.45 10.81 8.82 Sub-Total 6.45 18.92 13.23 Birds 12.90 8.11 10.3 Domestic Prey Poultry birds (Gallus gallus domesticus) 3.23 8.11 5.88 Sheep (Ovis aries) 6.45 2.70 4.41 Goat (Capra hircus) - 5.41 2.94 Dog (Canis familiaris) - 5.41 2.94 Sub-Total 9.68 21.63 16.17 Plants consumed Amaltas (Cassia fistula) 3.23 0.00 1.47 Peach (Prunus persica) 9.68 0.00 4.41 Shareen (Albizzia lebbek) 6.45 0.00 2.94 Yarrow (Achillea millefolium) 3.23 5.41 4.41 Daisy (Bellis perennis) - 56.76 30.9 Bindweed (Convolvulus arvensis) 3.23 2.70 2.94 Chandan (Chenopodium ambrosioides) - 2.70 1.47 Bair (Zizyphus muritiana) - 8.11 4.41 Bitter apple (Solanum incanum) - 2.70 1.47 Wild pomegranate (Punica granatum) - 8.11 4.41 Alu Bokhara (Prunus bokhariensis royle) 9.68 - 4.41 Granda (Carissa opaca) - 8.11 4.41 Maize (Zea mays) - 2.70 1.47 Soil 16.13 - 7.35 Anthropogenic matter 19.35 5.41 11.8 *s = summer; *w = winter; *F = frequency of occurrence. page 8 of 12Zoological Studies 60:34 (2021) © 2021 Academia Sinica, Taiwan We confirmed scats collected from the field to belong to leopard cat species through molecular analysis, before analyzing them for determine the dietary habits of the species. The analysis of scats showed that the species consumes both wild prey (51.75%), domestic prey (7.69%) and plant matter (32.41%), along with some anthropogenic matter (3.50%) in the study area. Our results revealed presence of a variety of mammals in the dietary menu of the leopard cat, with a high intake of rodents, including five different species—house rat, house mouse, Indian gerbil, Indian musk shrew, and Indian mole rat. These findings are in line with Shehzad et al. (2012) who reported that the leopard cat has a diversified diet, dominated by house rat (in 68% processed scats). In the current study, we report that rodents make up the majority of the leopard cat’s diet year-round. Our results are in line with many previous studies (Austin et al. 2007; Fernandez and de Guia 2011; Shehzad et al. 2012). The leopard cat does not just feed on rodents, but also tree nesting birds (Nowell and Jackson 1996). In Ayubia National Park (ANP) and Chitral Gol National Fig. 4. Seasonal indices in prey species of the leopard cat in Margalla Hills National Park Islamabad, Pakistan. Fig. 3. Percentage frequency (%F) of food items (wild and domestic prey & plant matter) in the diet of the leopard cat occurring in MHNP. page 9 of 12Zoological Studies 60:34 (2021)