Molecular analysis for investigating dietary habits: genetic screening of prey items in scat and stomach contents of leopard cats Prionailurus bengalensis euptilurus

Abstract

Background: Among the Felidae recorded on the Korean Peninsula, the tiger Panthera tigris, leopard Panthera pardus, and lynx Lynx lynx are now endangered, while the leopard cat Prionailurus bengalensis, the remaining feline, is a globally threatened carnivore. Herein, we investigated the dietary habits of leopard cats by analyzing prey DNA in scat and stomach contents. We also tested whether prey DNA in scat samples collected from natural habitats could accurately identify prey species from stomach contents. Results: Following a visual analysis of stomach contents from 11 leopard cats killed on the roads, a molecular analysis of the cytochrome b gene of the mitochondrial genome of 56 subsamples of the stomach contents enabled the identification of 7 mammalian species, 1 bird species, and 1 amphibian species. In the analysis of several blind subsamples (e.g., bones) isolated from fecal samples, five prey species were identified using control markers in a denaturing gradient gel electrophoresis (DGGE) technique and an additional sequencing analysis. Conclusions: Our results suggest that the DGGE analysis can serve as a potential tool to study diets, raising the possibility of a non-invasive approach for studying dietary habits of leopard cats.

Molecular dietary analysis of two sympatric felids in the Mountains of Southwest China biodiversity hotspot and conservation implications

“…Most previous studies of the LPC diet were based on microscopic examination of food remains in faeces where generally <10 taxa were identified, often only to the family or higher taxonomic levels 20 21 23 , demonstrating the superior resolution power of the faecal DNA-based method. The number of vertebrate prey categories identified in the present study (37 taxa) considerably exceeds not only reports based on a traditional cloning-Sanger sequencing approach for the LPC diet in the same area (16 taxa) 35 and in South Korea (13 taxa) 25 , but also a study based on NGS analysis of LPC faeces collected in northern Pakistan (18 taxa) 50 . Technical advances represented by the NGS approach offer remarkably higher throughput and economy compared to traditional morphological analysis and sequencing methods, demonstrating the high detection sensitivity and assignment power of molecular-metabarcoding/NGS methods.…”

“…Although LPC is detected at a relatively high frequency by camera-trapping surveys in the study area 18 , the population status of the species is unclear. Analyses of faeces and stomach contents show that the main LPC prey items are rodents (mostly murids), supplemented with other small mammals, such as squirrels and shrews, birds, reptiles, amphibians and fish, across its range 19 20 21 22 23 24 25 26 . Little data are available on their dietary habits in most regions of China.…”

Ecological preferences of large carnivores in remote, high-altitude protected areas: insights from Buxa Tiger Reserve, India

“…Prey remains in carnivore faecal samples can be identified by several methods, including microscopic hair analysis and DNA-based species identification. Although more accurate, DNA analysis of prey remains in a large number of samples by next-generation sequencing (Shehzad et al, 2012) or denaturing gradient gel electrophoresis (Lee et al, 2013) is more costly and technique-intensive. Microscopy of hair samples provides reliable identification at least to genus, and in many cases to the species level.…”

“Guess Who’s Coming to Dinner”: Molecular Tools to Reconstruct multilocus Genetic Profiles from Wild Canid Consumption Remains

“…During the last decades, diet analyses of vertebrate predators have been extensively conducted through multidisciplinary approaches, mainly based on morphological and molecular identifications of their prey remains contained in non-invasively collected faecal materials [ 4 , 5 ]. In particular, most molecular studies on diet have been focused on a simple taxonomic identification of the consumed species [ 6 , 7 ] using vertebrate broad range markers, such as the cytochrome b and the 16S subunit of the mitochondrial DNA, or applying metabarcoding techniques, to genotype prey DNA contained in predator scats [ 7 , 8 , 9 ]. However, the recent spread of anthropogenic hybridisation, which originated from the crossing between native and alien species or wild and domestic populations of the same species, requires the application of more powerful molecular tools.…”