Noninvasive Genetic Methods for Species Identification and Dietary Profiling of the Japanese Dormouse Glirulus japonicus from Fecal Samples

Noninvasive methods for species identification and dietary profiling from fecal samples in an artificial nest box were developed for the Japanese dormouse Glirulus japonicus. The species is a natural monument in Japan protected by domestic regulations since 1975. We assessed the mitochondrial cytochrome b gene for species identification and obtained location-specific sequences for Oki and Yamanashi, Japan. This marker was able to identify the dormouse species from feces. We also performed DNA metabarcoding analyses to clarify the invertebrate and plant diets of the Japanese dormouse. Various invertebrates and plants were detected, supporting the omnivorous nature of this species. Furthermore, almost all dietary items were identified to the genus level (82.1% and 99.5% for invertebrates and plants, respectively). The dietary components in Yamanashi Prefecture suggested that the Japanese dormouse mainly consumed Lepidoptera and various plants in summer, and Diptera and Hemiptera (aphids) invertebrates and Actinidia sp. plants in autumn. The latter plants produce fruit in autumn, enabling the dormouse to accumulate fat before hibernation. We discuss the potential and pitfalls of the noninvasive method, including the necessity of local DNA databases, reliability of the global DNA database, sampling procedure to avoid contamination, and individual identification.