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1.
Sialyl oligosaccharides were separated from two samples of Japanese black bear milk by extraction with chloroform/methanol, gel filtration on Bio Gel P-2, ion exchange chromatography on DEAE-Sephadex A-50 and high-performance liquid chromatography (HPLC) on a TSK gel Amido-80 column. They were characterized by 1H-NMR spectroscopy. The structures of four sialyl oligosaccharides separated from the milk were the following:
Neu5Ac(α2-3)Gal(β1-4)Glc
Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-3) Gal(α1-3)Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6) Gal(β1-4)Glc
Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-3) Gal(α1-3)[Fuc(α1-2)]Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6) Gal(β1-4)Glc
Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-3)[Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc
Keywords: Japanese black bear milk; Milk oligosaccharides; Sialyl milk oligosaccharides; B antigen; α-Gal epitope; Ursidae; Ursus thibetanus japonicus; Japanese black bear  相似文献   

2.
Two trisaccharides, three tetrasaccharides, two pentasaccharides, one hexasaccharide, one heptasaccharide, one octasaccharide and one decasaccharide were isolated from polar bear milk samples by chloroform/methanol extraction, gel filtration, ion exchange chromatography and preparative thin-layer chromatography. The oligosaccharides were characterized by 1H-NMR as follows: the saccharides from one animal: Gal(alpha1-3)Gal(beta1-4)Glc (alpha3'-galactosyllactose), Fuc(alpha1-2)Gal(beta1-4)Glc (2'-fucosyllactose), Gal(alpha1-3)[Fuc(alpha1-2)]Gal(beta1-4)Glc (B-tetrasaccharide), GalNAc(alpha1-3)[Fuc(alpha1-2)]Gal(beta1-4)Glc (A-tetrasaccharide), Gal(alpha1-3)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc, Gal(alpha1-3)[Fuc(alpha1-2)]Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Gl c, Gal(alpha1-3)Gal(beta1-4)GlcNAc(beta1-3)[Gal(alpha1-3)Gal(beta1-4)Glc NAc(beta1-6)]Gal(beta1-4)Glc; the saccharides from another animal: alpha3'-galactosyllactose, Gal(alpha1-3)Gal(beta1-4)[Fuc(alpha1-3)]Glc, A-tetrasaccharide, GalNAc(alpha1-3)[Fuc(alpha1-2)]Gal(beta1-4)[Fuc(alpha1-3)]Glc (A-pentasaccharide), Gal(alpha1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Gl c, Gal(alpha1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)[F uc(alpha1-3)]Glc (difucosylheptasaccharide) and Gal(alpha1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)?Gal(alpha1-3) Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-6)?Gal(beta1-4)Glc (difucosyldecasaccharide). Lactose was present only in small amounts. Some of the milk oligosaccharides of the polar bear had alpha-Gal epitopes similar to some oligosaccharides in milk from the Ezo brown bear and the Japanese black bear. Some milk oligosaccharides had human blood group A antigens as well as B antigens; these were different from the oligosaccharides in Ezo brown and Japanese black bears.  相似文献   

3.
Morphological and genetic features of a new Hepatozoon species, Hepatozoon ursi n. sp., in Japanese black bear (Ursus thibetanus japonicus) were studied. Schizogonic developmental stages were observed in the lungs of Japanese black bears. The schizonts were sub-spherical in shape and 45.7+/-4.6 x 42.7+/-4.5 microm in size. Each mature schizont contained approximately 80-130 merozoites and 0-5 residual bodies. The merozoites were 7.0+/-0.7 x 1.8+/-0.3 microm in size. Intraleukocytic gametocytes were slightly curved, cigar-like in shape and had a beak-like protrusion at one end. The size of the gametocytes was 10.9+/-0.3 x 3.3+/-0.2 microm. The analyses of the18S rRNA gene sequences supported the hypothesis that H. ursi n. sp. is different from other Hepatozoon species. Mature Hepatozoon oocysts were detected in two species of ticks (Haemaphysalis japonica and Haemaphysalis flava) collected on the bears infected with H. ursi n. sp. Two measured oocysts were 263.2 x 234.0 microm and 331.8 x 231.7 microm, respectively. The oocysts contained approximately 40 and 50 sporocysts, respectively. The sporocysts were sub-spherical in shape and 31.2+/-2.5 x 27.0+/-2.9 microm in size. Each sporocyst contained at least 8-16 sporozoites, with the sporozoites being 12.2+/-1.4 x 3.5+/-0.5 microm in size. H. ursi n. sp. is the first Hepatozoon species recorded from the family Ursidae.  相似文献   

4.
将采自23 头成年圈养黑熊的精液,分别用3 种稀释液(Ⅰ:Tris - 乳- 果- 卵;Ⅱ:柠- 葡- 蔗- 卵;Ⅲ:Tris - 柠- 果- 葡- 卵)稀释并在4℃ 下保存,通过检测精液在不同稀释液稀释条件下的保存时间,筛选出最适稀释液用于精液的冷冻保存;从精液解冻后精子的活率、活力、畸形率、顶体完整率4 个指标,分别从3种冷冻保护剂(甘油3%、3.5% 、4% )、两种冷冻方法(两步冷冻法和自动冷冻法)两个方面进行了比较试验。结果表明:精子活力在0.3 以上时,稀释液Ⅲ保存时间为175.42 ± 3.04 h,显著高于稀释液Ⅰ和稀释液Ⅱ(P ﹤0.01),稀释液Ⅱ保存时间也明显高于稀释液Ⅰ (P ﹤0.01);含3.5% 甘油浓度的稀释液解冻后精子活率(41.75 ± 3.46% )、活力(32.63 ±5.27% )和顶体完整率(85.62 ± 4.58% )显著高于其他两组(P ﹤0.01),并且精子畸形率(29.32 ± 8.22% )明显低于其他两组(35.95 ± 8.04% ,36.07 ±7.72% )(P ﹤0.01);采用自动冷冻法冷冻保存圈养黑熊精液,解冻后精子活率、活力和顶体完整率分别为41.75 ±3.46% 、32.63 ± 5.27%和85.62 ±4.58% ,都明显高于两步冷冻法(P ﹤0.01);解冻后畸形率为29.32 ± 8.22% ,明显低于两步冷冻法(P ﹤0.01)。  相似文献   

5.
6.
Ten polymorphic microsatellite markers were developed for the endangered Formosan black bear (Ursus thibetanus formosanus) from a partial genomic library enriched for GAAA repeat. Polymorphism of these loci was evaluated in 27 Formosan black bear specimens of unknown relationship. The number of alleles per locus ranged from 5 to 15 and the observed heterozygosity of each locus ranged from 0.556 to 0.889. These loci should provide useful molecular tools to study conservation genetics of the Formosan black bear and other Asiatic black bears.  相似文献   

7.
We evaluated the stored body fat of Japanese black bears (Ursus thibetanus japonicus) killed as nuisances in Gifu and Fukushima prefectures, Japan, during 2005-2007. We employed femur marrow fat (FMF), modified kidney fat index (mKFI), and abdominal subcutaneous fat (ASF) as indices for quantitative evaluation. We examined the basic characteristics of these indices, such as seasonality, age and sex dependency, and the quantitative relationship among them. mKFI and ASF increased towards the beginning of the denning period (December), while FMF was relatively stable throughout the sampling period (July-December). In cubs, all indices showed significantly lower values than in the older age classes. There seemed to be a catabolizing order between FMF and mKFI, but not between mKFI and ASF. We also evaluated the yearly change in the indices, and discussed its relevance to the incidence of bear intrusion into human residential areas. Bears nuisance-killed in summer (July-September) 2006 had a significantly larger amount of stored body fat than those killed in summer 2007, although the number of nuisance kills was larger in 2006 than in 2007. This suggests that poor nutritional condition is not a direct cause of bear intrusion.  相似文献   

8.
Two Dirofilaria ursi, six Trichodectes pinguis and numerous Haemaphysalis megaspinosa were obtained from an Asiatic black bear (Ursus thibetanus) shot in Kyushu, Japan. The presence of two parasites specific to black bears may indicate the existence of a wild population, although the bears are regarded to be extinct on Kyushu Island.  相似文献   

9.
This study characterized semen collected from the Japanese black bear, Ursus thibetanus aponicus, to provide information on semen cryopreservation for artificial breeding. Preliminary studies using a beagle dog as the model species showed that sperm concentration and total sperm count were lower in semen collected by electroejaculation than in semen collected by digital manipulation, but that sperm motility, viability and morphology were similar. Characterization of semen obtained from Japanese black bears by electroejaculation under general anesthesia revealed that semen volume and total number of spermatozoa collected were lower; but that sperm concentration, motility, viability and morphology were equivalent to those reported in other ursids. When semen was collected via a catheter inserted into the urethra during the stimulation for ejaculation, the sperm concentration, total sperm count and motility were relatively higher than when semen was collected directly in a test tube. Specific normal semen characteristics (mean +/- SEM) were pH, 7.6 +/- 0.0; volume, 0.212 +/- 0.038 mL; sperm concentration, 361 +/- 100 x 10(6)/mL; total sperm count, 84.0 +/- 32.2 x 106; +++ motility, 30 +/- 5%; motility, 77 +/- 3%; viability 77 +/- 2%; and abnormal morphology, 11+/- 2%. These results suggest that semen can be collected from Japanese black bears by electroejaculation.  相似文献   

10.
Trichinellosis is a meat-borne zoonotic disease caused by nine Trichinella speices and three unclassified genotypes. In Japan, four domestic outbreaks of human trichinellosis are reported sporadically and were associated with the consumption of wild bear meat. This study examined Trichinella prevalence and its species in black bears, Ursus thibetanus japonicus in Iwate prefecture, Japan. Trichinella T9 larvae identified molecularly were first detected in 1.4% (2/144) of the masseters of black bears examined, and their densities were low (1 and 0.3 larvae /g muscle, respectively). Two cytochrome C oxidase I (COI) haplotypes (sequences) of Trichinella T9 were found in distinct bear populations, suggesting that Trichinella T9 populations isolated genetically by bear populations would occur in Japan.  相似文献   

11.
Two trisaccharides, three tetrasaccharides, two pentasaccharides, one hexasaccharide, one heptasaccharide, one octasaccharide and one decasaccharide were isolated from polar bear milk samples by chloroform/methanol extraction, gel filtration, ion exchange chromatography and preparative thin-layer chromatography. The oligosaccharides were characterized by 1H-NMR as follows: the saccharides from one animal: Gal(α1-3)Gal(β1-4)Glc (α3′-galactosyllactose), Fuc(α1-2)Gal(β1-4)Glc (2′-fucosyllactose), Gal(α1-3)[Fuc(α1-2)]Gal(β1-4)Glc (B-tetrasaccharide), GalNAc(α1-3)[Fuc(α1-2)]Gal(β1-4)Glc (A-tetrasaccharide), Gal(α1-3)Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc, Gal(α1-3)[Fuc(α1-2)]Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc, Gal(α1-3)Gal(β1-4)GlcNAc(β1-3)[Gal(α1-3)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc; the saccharides from another animal: α3′-galactosyllactose, Gal(α1-3)Gal(β1-4)[Fuc(α1-3)]Glc, A-tetrasaccharide, GalNAc(α1-3)[Fuc(α1-2)]Gal(β1-4)[Fuc(α1-3)]Glc (A-pentasaccharide), Gal(α1-3)Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-3)Gal(β1-4)Glc, Gal(α1-3)Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-3)Gal(β1-4)[Fuc(α1-3)]Glc (difucosylheptasaccharide) and Gal(α1-3)Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-3){Gal(α1-3)Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6)}Gal(β1-4)Glc (difucosyldecasaccharide). Lactose was present only in small amounts. Some of the milk oligosaccharides of the polar bear had α-Gal epitopes similar to some oligosaccharides in milk from the Ezo brown bear and the Japanese black bear. Some milk oligosaccharides had human blood group A antigens as well as B antigens; these were different from the oligosaccharides in Ezo brown and Japanese black bears.  相似文献   

12.
The population genetic status of the Asian black bear Ursus thibetanus in the western part of Japan was examined using microsatellite DNA markers. Although three fragmented populations have been recognized in this region, i.e., the western Chugoku (WC), eastern Chugoku (EC), and northern Kinki (NK) populations, it was recently pointed out that the NK population has also been divided into two subpopulations, the western (WNK) and the eastern (ENK) populations. We examined the genetic status of these four populations based on their heterozygosities and the genetic distances (F ST ) between them. The mean observed heterozygosity, i.e., genetic diversity, of the six microsatellite loci examined, was significantly lower in WC (0.272; n = 52) and WNK (0.311; n = 66) than that in ENK (0.445; n = 67), respectively. Reflecting geographic distances among populations, F ST was higher between WC and WNK (0.244) and between WC and ENK (0.229) than that between WNK and ENK (0.066), respectively. There was also low but significant genetic differentiation between the two nearest populations (WNK and ENK). Although the status of EC was not conclusively determined because of its small sample size (n = 24), its genetic diversity seemed to be at a level similar to that of WC and WNK. These results suggest that WC, which is isolated from the others, has been losing genetic diversity, and that WNK, which had been thought to form a relatively large population together with ENK, might be isolated genetically from other populations. WNK, as well as WC and EC, should be conserved intensively. Received: August 2, 2001 / Accepted: September 20, 2001  相似文献   

13.
In this study, we tried to detect protozoan blood parasites from the liver or blood of 156 Japanese black bears (Ursus thibetanus japonicus) in Iwate Prefecture of Japan by polymerase chain reaction. Two amplicons (approximately 540 bp and 480 bp) were detected by amplification for V4 hyper-variable regions of the 18S rRNA gene. Approximately 540-bp products were obtained in 119 samples (76.3%) and were considered to be DNA of Hepatozoon ursi. Approximately 480-bp products were obtained in 22 samples (14.1%) and were considered to be DNA of Babesia species. The nucleotide sequences (1635 bp) of the 18S rRNA gene of Babesia sp. were very similar (99.3%) to those (AY190123, AY190124) of Babesia sp. detected previously from Ixodes ovatus. Phylogenetic analysis showed that Babesia sp. detected in this study closely related to Babesia sp. derived from raccoons in Japan and the U.S.A. This is the first report of Babesia species detected from Japanese black bears.  相似文献   

14.
We examined changes in the concentrations of serum progesterone (P4), estradiol-17beta (E2), FSH, LH, prolactin (PRL), and inhibin to determine their interaction and their effect on the reproductive endocrine controls of pregnant and nonpregnant female Japanese black bears. Fourteen female bears were used in this study over a 2-yr period. In the first year, six of the bears were divided into two groups; a pseudopregnant group and a nonpregnant group. In the second year, the remaining eight bears were also divided into two groups; a pregnant group and a nonpregnant group. Pregnant and pseudopregnant bears had similar P4 trends with both groups exhibiting a significant increase in December, which is the suspected time of implantation in pregnant bears. These trends correlated with an increase in PRL levels, whereas low levels of LH were maintained throughout the year. Nonpregnant bears maintained low concentrations of P4, and compared with pregnant and pseudopregnant bears, they also exhibited a delayed elevation in PRL. Luteinizing hormone activity varied among individual animals, but regardless of reproductive status, fluctuation patterns of E2, FSH, and inhibin did not differ among bears. Our results suggest that PRL may play a luteotropic role in both pregnant and pseudopregnant bears, and is possibly responsible for inducing reactivation of the dormant corpus luteum that precedes implantation in the Japanese black bear.  相似文献   

15.
We obtained the complete mitochondrial genome of U.thibetanus mupinensis by DNA sequencing based on the PCR fragments of 18 primers we designed. The results indicate that the mtDNA is 16,868 bp in size, encodes 13 protein genes, 22 tRNA genes, and 2 rRNA genes, with an overall H-strand base composition of 31.2% A, 25.4% C, 15.5% G and 27.9% T. The sequence of the control region (CR) located between tRNA-Pro and tRNA-Phe is 1422 bp in size, consists of 8.43% of the whole genome, GC content is 51.9% and has a 6bp tandem repeat and two 10bp tandem repeats identified by using the Tandem Repeats Finder. U. thibetanus mupinensis mitochondrial genome shares high similarity with those of three other Ursidae: U. americanus (91.46%), U. arctos (89.25%) and U. maritimus (87.66%).  相似文献   

16.
Developing a better understanding of the reproductive physiology and breeding condition peculiar to wild Japanese black bears (Ursus thibetanus japonicus) is crucial for estimation of their habitat distribution. The aim of this study was to clarify the changes in morphology of the genital organs, cellular proliferation in the endometrium and sex steroid hormone concentrations along with the reproductive cycle in Japanese black bears. Samples were collected from a total of 24 female Japanese black bears (1-15 presumptive years old) that were caught in the wild in Iwate prefecture during the period between August 1999 and September 2005. Twenty-two out of the 24 animals were hunted from May to October. The ovaries from the 24 animals and the uteri from 23 animals were observed macroscopically and histologically to examine the relationship between morphology of the genital organs and the month of the year the animal was caught. The staining pattern of proliferating cell nuclear antigen (PCNA) in the endometrium was characterised. Peripheral concentrations of oestradiol-17beta and progesterone were determined by radioimmunoassay. All the animals that had a corpus luteum (n=12) were captured from August to October. The thickness of the endometrium in the animals captured from August to October (n=16) was significantly greater than those in animals captured from May to July (n=5) (P<0.05). From August to October, the thickness of the endometrium and the ratio of the area of the uterine glands to the area of the endometrium in the animals with a corpus luteum (n=12) were significantly greater than those without a corpus luteum (n=4) (P<0.01). Positive PCNA staining was only observed in the uteri of two animals captured in May. There was a significantly positive correlation between plasma progesterone concentrations and the thickness of the endometrium (rho=0.589, P<0.05). There were also significantly positive correlations between the progesterone to oestradiol-17beta ratio (P4/E2 ratio) and the thickness of the endometrium (rho=0.710, P<0.05), and between the P4/E2 ratio and the ratio of the uterine gland area in the endometrium (rho=0.626, P<0.01). These data suggest that the corpus luteum is formed during or just after the breeding season and that the cells in the endometrium and the uterine glands, which proliferate in the early breeding season, grow and develop under the influence of progesterone and oestradiol-17beta during the period of delayed implantation in Japanese black bears.  相似文献   

17.
18.
中国西南山地是亚洲黑熊(Ursus thibetanus)的重要分布地,但是对该物种的活动模式研究很少.2017年7月-2019年8月,本研究以云南兰坪云岭省级自然保护区拉沙山区域亚洲黑熊为研究对象,按照250 m×250 m网格布设218台红外相机,累计有效拍摄日为36 175 d,共获得亚洲黑熊的独立照片(Inde...  相似文献   

19.

The milk oligosaccharides were studied for two species of the Carnivora: the American black bear (Ursus americanus, family Ursidae, Caniformia), and the cheetah, (Acinonyx jubatus, family Felidae, Feliformia). Lactose was the most dominant saccharide in cheetah milk, while this was a minor saccharide and milk oligosaccharides predominated over lactose in American black bear milk. The structures of 8 neutral saccharides from American black bear milk were found to be Gal(β1–4)Glc (lactose), Fuc(α1–2)Gal(β1–4)Glc (2′-fucosyllactose), Gal(α1–3)Gal(β1–4)Glc (isoglobotriose), Gal(α1–3)[Fuc(α1–2)]Gal(β1–4)Glc (B-tetrasaccharide), Gal(α1–3)[Fuc(α1–2)]Gal(β1–4)[Fuc(α1–3)]Glc (B-pentasaccharide), Fuc(α1–2)Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–3)Gal(β1–4)Glc (difucosyl lacto-N-neotetraose), Gal(α1–3)Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–3)Gal(β1–4)Glc (monogalactosyl monofucosyl lacto-N-neotetraose) and Gal(α1–3)Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc (Galili pentasaccharide). Structures of 5 acidic saccharides were also identified in black bear milk: Neu5Ac(α2–3)Gal(β1–4)Glc (3′-sialyllactose), Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)[Fuc(α1–2)Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (monosialyl monofucosyl lacto-N-neohexaose), Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)[Gal(α1–3)Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (monosialyl monogalactosyl lacto-N-neohexaose), Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3){Gal(α1–3)Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–6)}Gal(β1–4)Glc (monosialyl monogalactosyl monofucosyl lacto-N-neohexaose), and Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3){Gal(α1–3)[Fuc(α1–2)]Gal(β1–4)[Fuc(α1–3)]GlcNAc(β1–6)}Gal(β1–4)Glc (monosialyl monogalactosyl difucosyl lacto-N-neohexaose). A notable feature of some of these milk oligosaccharides is the presence of B-antigen (Gal(α1–3)[Fuc(α1–2)]Gal), α-Gal epitope (Gal(α1–3)Gal(β1–4)Glc(NAc)) and Lewis x (Gal(β1–4)[Fuc(α1–3)]GlcNAc) structures within oligosaccharides. By comparison to American black bear milk, cheetah milk had a much smaller array of oligosaccharides. Two cheetah milks contained Gal(α1–3)Gal(β1–4)Glc (isoglobotriose), while another cheetah milk did not, but contained Gal(β1–6)Gal(β1–4)Glc (6′-galactosyllactose) and Gal(β1–3)Gal(β1–4)Glc (3′-galactosyllactose). Two cheetah milks contained Gal(β1–4)GlcNAc(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (lacto-N-neohexaose), and one cheetah milk contained Gal(β1–4)Glc-3’-O-sulfate. Neu5Ac(α2–8)Neu5Ac(α2–3)Gal(β1–4)Glc (disialyllactose) was the only sialyl oligosaccharide identified in cheetah milk. The heterogeneity of milk oligosaccharides was found between both species with respect of the presence/absence of B-antigen and Lewis x. The variety of milk oligosaccharides was much greater in the American black bear than in the cheetah. The ratio of milk oligosaccharides-to-lactose was lower in cheetah (1:1–1:2) than American black bear (21:1) which is likely a reflection of the requirement for a dietary supply of N-acetyl neuraminic acid (sialic acid), in altricial ursids compared to more precocial felids, given the role of these oligosaccharides in the synthesis of brain gangliosides and the polysialic chains on neural cell adhesion.

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