Combining vertebrate mitochondrial 12S rRNA gene sequencing and shotgun metagenomic sequencing to investigate the diet of the leopard cat (Prionailurus bengalensis) in Korea

Abstract

The leopard cat (Prionailurus bengalensis), an endangered species in South Korea, is a small feline widely distributed in Asia. Here, we investigated the diet of leopard cats in the inland areas of Korea by examining their fecal contents using vertebrate mitochondrial 12S rRNA gene sequencing and shotgun metagenomic sequencing. Shotgun metagenomic sequencing revealed that the feces were rich in DNA not only of vertebrates but also of arthropods and plants, but care should be taken when using shotgun metagenomic sequencing to identify vertebrates at low taxonomic levels (e.g., genus level), as it was often erroneous. Meanwhile, vertebrate mitochondrial 12S rRNA gene sequencing was found to be accurate in the genus-level identification, as the genera identified were consistent with the Korean fauna. We found that small mammals such as murids were their main prey. By using these two sequencing methods in combination, this study demonstrated that accurate information about the overall dietary content and vertebrate prey of leopard cats could be obtained. We expect that the continued community efforts to expand the genome database of wildlife, including vertebrates, will alleviate the problem of erroneous identification of prey at low taxonomic levels by shotgun metagenomic sequencing in the near future.

Diversity of fecal parasitomes of wild carnivores inhabiting Korea, including zoonotic parasites and parasites of their prey animals, as revealed by 18S rRNA gene sequencing

“…Few cases of the detection of T. vulpis from felines have been reported, and we do not know the reason. One hypothesis is that the leopard cat concomitantly preyed on the dog parasitized with T. vulpis , because our dietary study identified the dog as a dietary item of leopard cats in our study areas ( Woo et al, 2023 ).…”

“…In addition, in the feces of leopard cats, genera containing species whose intermediate and/or definitive hosts are small mammals such as rodents and/or birds, e.g., the apicomplexa Eimeria ( Todd and Lepp, 1971 ) and Sarcocystis ( Prakas et al, 2019 ), the nematode and Heterakis ( Smith, 1953 ; Katherine Lynn and Robert Byron, 2019 ), and the platyhelminth Neodiplostomum ( Sanmartin et al, 2004 ), were abundantly detected. Our dietary study revealed that murids and birds are the main prey of leopard cats in the inland areas ( Woo et al, 2023 ). It is well known that parasites pass from prey to predators, in which they are digested to death or an infection is established ( Thieltges et al, 2013 ).…”

“…Fecal samples previously collected for diet studies of the wild animals ( Kumari et al, 2019 ; Woo et al, 2022 , 2023 ) were used. A total of 40 fecal samples were collected, consisting of 11, 22, and 7 samples of raccoon dogs, leopard cats, and Eurasian otters, respectively.…”

“…A total of 40 fecal samples were collected, consisting of 11, 22, and 7 samples of raccoon dogs, leopard cats, and Eurasian otters, respectively. Sample metadata are listed in Supplementary Table 1 and sample collection methods are reported in our previous studies ( Kumari et al, 2019 ; Woo et al, 2022 , 2023 ). Briefly, fecal samples of raccoon dogs and Eurasian otters were collected from a waterside area ( Woo et al, 2022 ) and an estuary area ( Kumari et al, 2019 ), respectively.…”

“…Briefly, fecal samples of raccoon dogs and Eurasian otters were collected from a waterside area ( Woo et al, 2022 ) and an estuary area ( Kumari et al, 2019 ), respectively. Fecal samples of leopard cats were collected in inland areas ( Woo et al, 2023 ). Approximately 10 g of each sample was collected with a sterile wooden spatula in the field.…”