Fecal steroid metabolites and reproductive monitoring in a female Tsushima leopard cat (Prionailurus bengalensis euptilurus)

Although the Tsushima leopard cat (Prionailurus bengalensis euptilurus) is one of the most endangered mammals in Japan, its reproductive physiology and endocrinology have been not elucidated. The objective was to establish the non-invasive monitoring of reproductive endocrinology in a female Tsushima leopard cat and to identify the types of fecal reproductive steroid metabolites in this species. Fecal concentrations of estrogen and progestin were determined by enzyme immunoassays, from 60 d before to 60 d after the last copulation, during three pregnancies. Fecal estrogen metabolite concentrations were increased before/around the mating period and after mid-pregnancy. Fecal progestin metabolite concentrations increased after the last copulation and remained high during pregnancy. The gestation period was 65.0 ± 0.6 d (mean ± SD). Fecal extracts were separated by high-performance liquid chromatography for identification of fecal metabolites. Fecal estrogens were identified as estradiol-17β and estrone. Fecal progestins during pregnancy contained 5α-reduced pregnanes: 5α-pregnan-3α-ol-20-one, 5α-pregnan-3β-ol-20-one and 5α-pregnan-3,20-dione, and nonmetabolized progesterone was barely detected in feces. In conclusion, measurement of fecal estrogen and progestin metabolites was effective for noninvasive reproductive monitoring in the Tsushima leopard cat. An immunoassay for fecal estradiol-17β concentrations seemed useful to monitor follicular activity, whereas an immunoassay with high cross reactivity for 5α-reduced pregnanes was useful to monitor ovarian luteal activity and pregnancy.     

A cost-effective blood DNA methylation-based age estimation method in domestic cats,Tsushima leopard cats (Prionailurus bengalensis euptilurus) and Panthera species,using targeted bisulphite sequencing and machine learning models

Individual age can be used to design more efficient and suitable management plans in both in situ and ex situ conservation programmes for targeted wildlife species. DNA methylation is a promising marker of epigenetic ageing that can accurately estimate age from small amounts of biological material, which can be collected in a minimally invasive manner. In this study, we sequenced five targeted genetic regions and used 8–23 selected CpG sites to build age estimation models using machine learning methods at only about $3–7 per sample. Blood samples of seven Felidae species were used, ranging from small to big, and domestic to endangered species: domestic cats (Felis catus, 139 samples), Tsushima leopard cats (Prionailurus bengalensis euptilurus, 84 samples) and five Panthera species (96 samples). The models achieved satisfactory accuracy, with the mean absolute error of the most accurate models recorded at 1.966, 1.348 and 1.552 years in domestic cats, Tsushima leopard cats and Panthera spp. respectively. We developed the models in domestic cats and Tsushima leopard cats, which were applicable to individuals regardless of health conditions; therefore, these models are applicable to samples collected from individuals with diverse characteristics, which is often the case in conservation. We also showed the possibility of developing universal age estimation models for the five Panthera spp. using only two of the five genetic regions. We do not recommend building a common age estimation model for all the target species using our markers, because of the degraded performance of models that included all species.     

Sighting frequency and food habits of the leopard tortoise, Geochelone pardalis, in northern Tanzania

Sighting frequency and food habits of the leopard tortoise (Geochelone pardalis) were studied in northern Tanzania from October 1993 to June 1996. Sighting frequency varied significantly between protected (0.22 mhr^?1) and unprotected sites (0.59 mhr^?1), and between Arusha (0.27 mhr^?1) and Serengeti sites (0.56 mhr^?1). The tortoise diet comprised mostly plants (97.8%) and rarely inorganic matter (2.2%). A total of 47 plant species from 21 families was eaten. Forbs made up 74.5% of the plant items and monocots the remainder. Succulents contributed 51.0% of the plant items eaten while grasses and legumes accounted for 16.8% and 13.5%, respectively. Some plant items were eaten more frequently than their occurrence in the habitats.     

泰国Khao Yai国家公园豹猫和云豹的生态和保护

于1997年10月到1999年10月在泰国KhaoYai国家公园对两种同域分布的物种（即豹猫和云豹）进行了研究。使用带诱饵的陷阱捕捉了这些动物,进行麻醉,确定了性别、年龄,并戴上了无线电项圈,对6只雌性豹猫和4只雄性豹猫进行了1—18个月的无线电追踪。豹猫在干旱季节的利用区域大于雨季,雄性利用的区域大于雌性。在夜间和晨昏时节,豹猫活动增加,但是并没有节律性活动。所有豹猫个体在旱季和雨季的活动都相似,但雄性个体的白天活动多于雌性。豹猫的行走距离有性别差异但是没有季节差异;其食物以鼠类为主。分别对1只雌性云豹和1只雄性云豹进行了17个月和7个月的无线电追踪,雌性个体的活动面积为39．4km^2,而雄性个体活动面积为42．2km^2,核心区都是2．9km2。云豹对半绿林的使用大于其它类型的植被,在晨昏和夜间的活动增加,其活动无节律。     

Seasonal change in the food habits of the Iriomote catFelis iriomotensis

The food habits of the Iriomote catFelis iriomotensis were studied by analyzing 177 feces collected monthly from 1987 to 1988. A total of 26 food items were identified. The frequency of lizards and frogs in the feces was higher than those of mammals and birds. The food habits changed greatly seasonally.Eumeces skinks were fed on most frequently, and found in the feces with a similar high frequency occurrence in March–April and July–September, while their proportion to the total number of food items was larger in March–April than in July–September. The cats fed on larg-sized skinks, adultEumeces kishinouyei, more in March–April than in other seasons. The number of skinks sighted in the course of a road census was greater from March to August, and large-sized skinks were sighted more in March–April than in July–August. The cats fed selectively on large-sized skinks in every season. Therefore, changes in the food habits depended on the food availability. Characteristics of food habits in the Iriomote cat are discussed in comparison with the food habits of other felids in temperate and tropical regions.     

Long-term variation in fruiting and the food habits of wild Japanese macaques on Kinkazan Island, northern Japan

We studied the relationship between the food habits of Japanese macaques (Macaca fuscata) and the availability of nuts (Fagus crenata, Zelkova serrata, Carpinus spp., and Torreya nucifera) on Kinkazan Island, northern Japan, from 1997 to 2005 to examine the long-term variations in both food habits and availability. The food habits of the monkeys showed clear seasonal changes: the staple foods were woody leaves and flowers in spring (May and June), woody leaves and seeds or fruits other than nuts in summer (July and August), nuts and seeds or fruits other than nuts in fall (September-November), and herbaceous plants in winter (December-April). The availability of nuts, combinations of masting species, and energy production varied among years. Food habits varied among years, but the magnitude of variability of food habits differed among seasons, with large variability during summer and winter, and small variability in spring. Food availability was poor in summer and winter, but in several years the monkeys were able to consume nuts during those seasons. We emphasize the importance of conducting long-term studies on both food availability and the food habits of animals in the temperate zone.     

Diet and prey profiles of three sympatric large carnivores in Bandipur Tiger Reserve, India

We conducted a field study of diets of three sympatric large carnivores, the tiger Panthera tigris , the leopard Panthera pardus and the dhole Cuon alpinus in Bandipur Tiger Reserve, India, based on analyses of 381, 111 and 181 scats, respectively. The frequency of occurrence of prey items in scats was converted to relative biomass and number of prey consumed using regression equations based on earlier feeding trials. The results showed that although these predators kill ∼11–15 species of vertebrate prey, relatively abundant ungulate species provide 88–97% of biomass consumed by them. Although the dietary niche overlap among the three species was high (Pianka's index of 0.75–0.93), some specialized predation was observed. The largest ungulates, gaur Bos gaurus and sambar Cervus unicolor , provided 73% of biomass consumed by tigers, whereas medium-sized chital Axis axis and wild pig Sus scrofa formed 65 and 83% of the biomass intake of leopards and dholes, respectively. In terms of the relative numbers of prey animals killed by the three predators, chital, which is the most abundant prey species, dominated their diets (tiger=33%, leopard=39% and dhole=73%). The results of the study, in conjunction with earlier work, support the prediction that abundance of ungulate prey species, as well as their availability in different size classes, are both critical factors that facilitate sympatry among the three predators.     

Use of mires and food habits of sika deer in the Oze Area,central Japan

Sika deer Cervus nippon entered the Oze Area, a snowy area in central Japan, in the 1990s. The use of mires and the food habits of this species were studied in 1999 and 2000. Deer used the mires immediately after snow melted and did not use the mires in mid summer, although they did use them lightly in autumn. The summer food habits differed from those of the deer population living in Nikko in the lower area: non-Sasa graminoids and other monocotyledonous leaves occupied more (approximately 30%) of the feces in the former population, whereas Sasa nipponica, a dwarf bamboo, was the dominant food (60%) of the latter population. Seasonal changes in fecal composition in the Oze deer were that mire graminoids occupied a considerable portion of their food in spring (11.6%) and summer (17.0%), whereas the amount of dwarf bamboo increased in fall (26.0%). Despite the small size of the mires, Oze deer appear to prefer mires to forests. Nitrogen concentrations of the mire plants did not differ from those of the forest plants; however, foraging efficiency would be greater in the mires and this may explain the preference observed. Despite the small total biomass, leaf biomass of the mire was equivalent to that of the adjacent forest floor. In addition, biomass was concentrated near the ground, or the biomass concentration was greater than that of the forest floor.     

Geographic Variations in Dorsal and Anal Ray Counts of Juvenile Japanese Flounder, Paralichthys olivaceus, in the Japan Sea

Meristic counts of early and late wild settled juveniles of Japanese flounder, Paralichthys olivaceus (Paralichthyidae), in Wakasa Bay, Japan, were compared, with specimens collected from 26 sites spread along ca. 1700km of coastline facing the Japan Sea. Mean dorsal and anal ray counts of the early settled group at Kyoto were significantly larger than those of the late settled group. A discontinuous geographic boundary in mean dorsal and anal ray counts was found around Wakasa Bay and the Noto Peninsula with both means significantly larger in the southern group than in the northern group. Mean ray counts of the early and late settlers at Kyoto were similar to those of the southern and northern groups, respectively. The effect of water temperature on meristic characters was tested on two groups of laboratory-reared juveniles. Mean dorsal and anal ray counts were larger in juveniles reared at higher water temperatures. The early and late settler groups were estimated to hatch in late February and late April, respectively, representing negative response of meristic characters to water temperatures if these two groups hatched out in Wakasa Bay. Differences in the dorsal and anal ray counts between the early and late settled groups at Kyoto can, therefore be attributed to genotypic, not phenotypic variation. This suggests the possibility of transport of larvae of the early group from areas further south to Wakasa Bay by the warm Tsushima Current, a branch of the Kuroshio, which flows northeastward along the coast of the Japan Sea, particularly when the temperature gradation between the south and north areas during these seasons is considered.     

Seasonal food habits of a sika deer population in the warm temperate forest of the westernmost part of Honshu, Japan

The rumen contents of 63 sika deer (Cervus nippon Temminck) shot in the westernmost part of Honshu were analyzed by the point-frame method. The rumen contents were dominated by forbs and the majority of these were evergreen monocotyledonous plants, probably Ophiopogon spp. Evergreen browse leaves (woody leaves), such as Eurya japonica Thunb., Ilex crenata Thunb. and Trachelospermum asiaticum Nakai, were also important, constituting 20–40% of the rumen contents. Other categories accounted for only small proportions, although in summer, graminoids increased but twigs and bark decreased in importance and in fall and winter, fruits and seeds increased in importance. The diet of the deer was characterized by high-quality foods even in winter and by a stability or less-pronounced seasonality throughout the year than found in the northern deer. These results were compared with those from other populations, including a northern sika deer population in Japan.     

韩国豹猫与其它豹猫及西表猫之间的遗传分化:基于IRBP、12S rRNA 和Cyt b 序列的分析

利用韩国豹猫的IRBP、12S rRNA 和细胞色素b (Cyt b)序列,并将其与GenBank 中的豹猫及西表猫的上述基因进行比较,以此来重新分析韩国豹猫和其它豹猫及西表猫之间的遗传分化情况。首先,我们从18 只韩国豹猫的IRBP、12S rRNA 和Cyt b 序列分析中分别检测到6 个等位基因、1 个单倍型和3 个单倍型,推测曾经发生 在豹猫种群的瓶颈事件可能是本文发现的不同分子标记之间存在基因型丢失差异的原因。基于Cyt b 序列分析,证实东亚的豹猫形成一个单独的进化枝,本文得到的Cyt b 序列和之前发布结果支持一个分类理论,即认为东亚的豹猫就是东北森林猫。我们发现在IRBP、12S rRNA 基因上,韩国豹猫和西表猫存在相同的序列,支持将西表猫和豹猫视为同一物种。     

Linking genetic diversity and temporal fluctuations in population abundance of the introduced feral cat (Felis silvestris catus) on the Kerguelen archipelago

Linking temporal variations of genetic diversity, including allelic richness and heterozygosity, and spatio-temporal fluctuations in population abundance has emerged as an important tool for understanding demographic and evolutionary processes in natural populations. This so-called genetic monitoring was conducted across 12 consecutive years (1996-2007) at three sites for the feral cat, introduced onto the Kerguelen archipelago fifty years ago. Temporal changes in allelic richness and heterozygosity at 18 microsatellite DNA loci were compared with temporal changes in the adult population abundance index, obtained by typical demographic monitoring. No association was found at the island spatial scale, but we observed an association between genetic diversity and adult population indices from year to year within each study site. More particularly, the magnitude of successive increases or decreases in the adult population abundance index appeared to be the major factor linking the trajectories of genetic diversity and adult population abundance indices. Natal dispersal and/or local recruitment, both facilitated by high juvenile survival when the adult population size is small, is proposed as the major demographic processes contributing to such an observed pattern. Finally, we suggested avoiding the use of the harmonic mean as an estimator of long-term population size to study the relationships between demographic fluctuations and heterozygosity in populations characterized by strong multiannual density fluctuations.     

内蒙古达赉湖地区和蒙古国东方省蒙原羚的夏季食性

2008年7—8月,收集了内蒙古达赉湖地区和蒙古国东方省蒙原羚的新鲜粪便,采用粪样显微组织学分析法研究了蒙原羚的夏季食性。研究结果如下:(1)内蒙古达赉湖地区夏季植物群落在植被盖度、地上生物量、植物种数方面与蒙古国东方省差异均不显著;在内蒙古达赉湖地区和蒙古国东方省共采集常见植物18科43属63种;(2)内蒙古达赉湖地区蒙原羚共采食10科19属22种植物,蒙古国东方省蒙原羚共采食11科24属29种植物;(3)禾本科植物是内蒙古达赉湖地区和蒙古国东方省蒙原羚夏季主要食物,分别占蒙原羚食物组成的79.1%和73.2%;禾本科植物中的羊草、针茅是内蒙古达赉湖地区和蒙古国东方省蒙原羚的主要食物,分别占蒙原羚食物组成的63.6?和57.7%;(4)内蒙古达赉湖地区蒙原羚夏季食物组成生态位宽度Bj=2.0,蒙古国东方省蒙原羚夏季食物组成生态位宽度Bj=2.3;(5)内蒙古达赉湖地区与蒙古国东方省蒙原羚的食物相似性指数PS=0.7;(6)内蒙古达赉湖蒙原羚与蒙古国东方省蒙原羚主要食物秩相关性检验显著相关(P0.05),相关性系数0.6。研究表明尽管内蒙古达赉湖地区蒙原羚主要在草原围栏内活动,但与蒙古国东方省蒙原羚的夏季食性显著相似,这表明有限的活动范围没有对蒙原羚夏季食性产生显著影响。     

Characterization,tissue distribution of resistin gene and the effect of fasting and refeeding on resistin mRNA expression in Siberian sturgeon (Acipenser baerii)

Resistin as an adipokine identified from rodents in 2001 is involved in many biological processes. However, little is known about this gene in fish. We cloned Siberian sturgeon (Acipenser baerii) resistin cDNA of 795 base pairs, encoding 107 amino acids, which showed 38–40% identity to human and rodents. Real-time quantitative PCR showed that the resistin gene was widely distributed in tissues of Siberian sturgeon, with the highest expression in liver. After fasting for 1, 3, 6 and 10 days, the expression of the resistin gene in the liver of Siberian sturgeon decreased significantly, and after refeeding on the 10 days of fasting the resistin mRNA expression increased rapidly, suggesting that resistin may play an important role in liver in response to starvation. Taken together, these results suggest that resistin may be involved in the regulation of energy homeostasis in liver.     

Nuclear microsatellite and mitochondrial DNA analyses reveal the regional genetic structure and phylogeographical history of a sanguivorous land leech,Haemadipsa japonica,in Japan

Recent molecular studies have indicated that phylogeographical history of Japanese biota is likely shaped by geohistory along with biological events, such as distribution shifts, isolation, and divergence of populations. However, the genetic structure and phylogeographical history of terrestrial Annelida species, including leech species, are poorly understood. Therefore, we aimed to understand the genetic structure and phylogeographical history across the natural range of Haemadipsa japonica, a sanguivorous land leech species endemic to Japan, by using nine polymorphic nuclear microsatellites (nSSR) and cytochrome oxidase subunit one (COI) sequences of mitochondrial DNA (mtDNA). Analyses using nSSR revealed that H. japonica exhibited a stronger regional genetic differentiation among populations (G'_ST = 0.77) than other animal species, probably because of the low mobility of land leech. Analyses using mtDNA indicated that H. japonica exhibited two distinct lineages (A and B), which were estimated to have diverged in the middle Pleistocene and probably because of range fragmentation resulting from climatic change and glacial and interglacial cycles. Lineage A was widely distributed across Japan, and lineage B was found in southwestern Japan. Analyses using nSSR revealed that lineage A was roughly divided into two population groups (i.e., northeastern and southwestern Japan); these analyses also revealed a gradual decrease in genetic diversity with increasing latitude in lineage A and a strong genetic drift in populations of northeastern Japan. Combined with the largely unresolved shallow polytomies from the mtDNA phylogeny, these results implied that lineage A may have undergone a rapid northward migration, probably during the Holocene. Then, the regional genetic structure with local unique gene pools may have been formed within each lineage because of the low mobility of this leech species.     

Bark stripping of hinoki cypress by sika deer in relation to snow cover and food availability on Mt Takahara,central Japan

Bark stripping of hinoki cypress (Chamaecyparis obtusa (Sieb. et Zucc.) Endl.) by sika deer (Cervus nippon Temminck) was monitored for 4years on approximately 300 trees at three sites on Mt Takahara, central Japan. We investigated the effects of snow cover and food availability on seasonal and yearly changes in bark stripping. Bark stripping occurred during snowy periods (January and February) when less food was available because of the snow cover. The most serious bark stripping was observed in 1996 when the snow was the deepest and food resources were the poorest among the 4years of the study. In contrast, bark stripping did not occur in 1999 when the snow was thin and, consequently, more food was available. These results suggest that snow cover and food avail-ability influence the occurrence and intensity of bark stripping both seasonally and annually.     

Influence of artificial food supply on population parameters and dispersal in the Hakone T troop of Japanese macaques

A free-ranging troop of Japanese monkeys was observed for seven years from April 1971 to March 1978 in the Hakone area, Kanagawa Prefecture, Japan. This troop was fed artificially between 1956 and 1977. The artificial food supply was reduced by about half after 1974. The troop-size fluctuated around 100 between 1971 and 1974, but fell drastically from 91 in 1975 to 58 in 1978. Population parameters were compared between the two periods of 1971–1974 and 1975–1977. Clear differences between the two periods were found in primiparous age, ratio of non-natal resident males to all resident males, ratio of newcomers to non-natal resident males, age-specific disappearance rate, and proportion of deserters observed in the study area. A correlation existed between the number of males and females of 5 or more years of age in the troop. The numbers in the three age-sex groups (natal males, non-natal males, and females of 5 or more years of age in the troop) were closely connected with one another. The scarcity of food supply may have caused not only males but also females to disperse. The drastic decrease in troop-size after 1974 could be based on the disappearance of adult females. The cause of the dispersal of young males and adult females could have been mutual competition among the troop members for food, and that in adult males could have been competition for females.