ABSTRACT. The Japanese black bear (Ursus thibetanus japonicus) is endangered for extinction in some areas of Japan, and semen collection and cryopreservation are an important means to preserve genetic resources. The aim of this study was to characterize and c ryopreserve semen of free-ranging Japanese black bears. Semen was collected by electroejaculation procedure from 4 free-ranging Japanese black bears at the capture point in the field. Ejaculates containing motile sperm were recovered from all of the animals and ejaculate volume, total sperm count, % motility (percentage of motile spermatozoa), % viability (percentage of spermatozoa that excluded eosin) and % abnormal morphology (range (mean)) were 0.65-2.20 (1.51) ml, 99-1082 (490) × 10 6 , 5-100 (31), 42-97 (66) and 20-87 (53), respectively. Three of the 4 ejaculates were diluted with an egg yolk-TRIS-citrate-glucose extender and cryopreserved in liquid nitrogen. Motile spermatozoa were observed after freezing and thawing in all cases. This study showed that electroejaculation would be a useful method for collecting semen from free-ranging Japanese black bears and that at least motile spermatozoa would be obtained by fre ezing the thus collected electroejaculates. KEY WORDS: cryopreservation, electroejaculation, free-ranging, Japanese black bear, semen.J. Vet. Med. Sci. 66(11): 1371-1376, 2004 The Japanese black bear (Ursus thibetanus japonicus) is a large mammal that inhabits the islands of Honshu and Shikoku in Japan. In recent years, their habitats have become fragmented and shrunken by human activity, and this species even faces extinction in some areas [7]. What is worse, bears have been killed to protect forestry, agriculture, livestock and humans from damage caused by them.Endangered and threatened wild animals are now attracting attention as a genetic resource and for species conservation, and semen has been recovered from various captive non-domestic animals, including fat-tailed dunnarts, koalas, brushtail possums, long-footed potoroos, northern brown bandicoots and ring-tailed possums [32] [4,20,22,24], Hokkaido brown bears [13,14] and Japanese black bears [16]. However, these studies were done with captive animals, and there are no reports on semen collection and cryopreservation in free-ranging bears. Cryopreservation of semen collected from free-ranging animals is one of the most important methods for preservation of genetic resources and useful for artificial breeding. Moreover, characteristics of semen from wild animals may be essential for understanding reproductive status under field conditions, although studies using captive animals, kept under controlled conditions, may provide valuable data to analogize state under field conditions. It is easy to expect that the state of captive animals may differ from that of free-ranging ones to some extent, because of differences in their living environment and diet. It is difficult to obtain semen from free-ranging Japanese black bears due to limited access to the animals and seasonality of their ...