黑龙江省熊类资源现状及其保护对策

黑龙江省现分布有 2种熊 ,棕熊 (Ursusarctoslasiotus)和黑熊 (Selenarctosthibetanusussuricus)。棕熊广泛分布于全省各个林区 ;黑熊在大兴安岭无分布 ,仅分布于小兴安岭和东部山地的张广才岭、老爷岭和完达山林区 ,其分布北界为北纬 48～ 49°,东经 1 2 8～ 1 2 9°附近。 2种熊的总分布区虽无明显变化 ,但集中分布区明显向大兴安岭、小兴安岭、东部山地的腹地退缩。 1 996～ 2 0 0 0年全省棕熊的分布数量为 (63 1±1 43 )头 (n =463 9) ,种群密度为 (0 0 0 2 2± 0 0 0 0 5 )头 km2 ;黑熊的分布数量为 (1 1 1 0± 2 93 )头 (n =463 9) ,种群密度为 (0 0 0 65± 0 0 0 1 7)头 km2 。全省森工国有林区熊类分布数量 (1 1 71± 2 98)头 (n =61 2 ) ,与1 992年该林区熊类调查结果 (3 0 5 7± 73 0头 ;n =2 1 3 6)相比较 ,全林区 6年间熊类数量减少了 1 886头 ,减少率达 61 7% ,年递减率为 1 6%。熊类资源呈锐减趋势。其主要原因为 :(1 )森林长期超量采伐引起熊类栖息生境总体质量下降 ;(2 )人类活动导致熊类栖息地片断化日趋严重 ;(3 )人为捕杀是造成熊类数量减少的直接原因。最后 ,提出了今后全省熊类保护管理的 5点建议。     

Evidence of a phylogeographic break in the Romanian brown bear (Ursus arctos) population from the Carpathians

We analysed 33 brown bears from the Romanian Carpathians and the Italian Apennines at sequences of the mitochondrial control region and nine polymorphic microsatellite loci with regard to genetic variability and haplotype distribution. The Italian brown bears were monomorphic for mtDNA sequences. The Romanian bears yielded the highest variability found so far in this species. Haplotypes of both previously identified mtDNA lineages (western and eastern) were found in Romania. In the eastern part of the Carpathians western and eastern haplotypes occurred sympatrically, the bears from the western part of the mountain range only exhibited western-type sequences. This pattern provides evidence of a mitochondrial phylogeographic break in the distribution of the eastern lineage within the Romanian Carpathians. Conservation implications of this finding are discussed.     

Quality of frozen-thawed semen in brown bear is not affected by timing of glycerol addition

We have tested several freezing protocols for brown bear semen, modifying the time when glycerol was added (before and after cooling to 5 °C). No differences were found among protocols, indicating a good tolerance of brown bear semen to glycerol. This finding indicates that freezing protocols for brown bear semen could be modified to fit practical solutions which would facilitate preparation of the seminal samples in the field with the addition of glycerol at ambient temperature.     

Effect of basic factors of extender composition on post-thawing quality of brown bear electroejaculated spermatozoa

The improvement of freezing extenders is critical when defining sperm cryopreservation protocols for wild species, in order to create germplasm banks. The aim of this study was to evaluate the effect of additives (Equex Paste and EDTA) supplementation, egg-yolk (10 and 20%) and glycerol (4 and 8%) concentrations and extender osmolality (300 and 320 mOsm/kg) on the post-thawing quality of brown bear semen. Semen was obtained from 20 adult males by electroejaculation, and centrifugated individually (600 × g for 6 min). The pellets were diluted 1:1 in the corresponding extender TTF (TES-Tris-Fructose with the aforementioned variants) and cooled to 5 °C. Then, it was diluted down to 100 × 10⁶ spz/mL, loaded in 0.25 mL straws and frozen at −20°C/min. After thawing (in water at 65 °C for 6s), the semen samples were assessed for motility (CASA), viability (SYBR-14 with propidium iodide), acrosomal status (PNA-FITC with propidium iodide) and mitochondrial activity (JC-1). Extender supplementation with additives rendered significantly higher results for these sperm parameters. Comparing the two percentages of egg yolk, 20% egg yolk showed the highest motility results, percentages of viable spermatozoa and viable spermatozoa with intact acrosome. No differences were detected among samples frozen using 4 or 8% glycerol. For extender osmolality, 300 mOsm/kg showed higher values of VAP, VCL, VSL, and ALH than 320 mOsm/kg. Based on the best performance of sperm motility, viability and acrosome status, we conclude that the most suitable extender to cryopreserve brown bear spermatozoa was TTF adjusted to 300 mOsm/kg, supplemented with 20% egg yolk, 4-8% glycerol, and the additives 1% Equex paste and 2% EDTA.     

Evaluation of the qualitative and quantitative effectiveness of three media of centrifugation (Maxifreeze, Cushion Fluid Equine, and PureSperm 100) in preparation of fresh or frozen-thawed brown bear spermatozoa

Centrifugation is a crucial procedure in sperm cryopreservation protocols of brown bear (Ursus arctos), because the semen must be processed to increase sperm concentration and/or clean urine-contaminated samples. The efficacy of three media for centrifugation (Maxifreeze [IMV technologies, L'Aigle, France], Cushion Fluid Equine (Minitübe, Tiefenbach, Germany), and PureSperm [Nidacon, Gothenburg, Sweden]) on the quality of bear spermatozoa was evaluated. In experiment one, two cushioned media used for protecting against mechanical stress during centrifugation were analyzed. In experiment two, a density gradient based on PureSperm was assessed in relation to the maximum retrieval and the quality of fresh spermatozoa, and the freezability of the spermatozoa selected in this density gradient was studied in experiment three. Finally, the selection of frozen-thawed sperm using PureSperm was analyzed in experiment four. Our results indicate that the use of dense isotonic cushion solutions (Maxifreeze, Cushion Fluid Equine) in centrifugation did not improve the quality of recovered spermatozoa compared with standard centrifugation. However, a density gradient prepared with PureSperm improved the quality of spermatozoa in fresh semen and frozen-thawed semen, but the spermatozoa selected from the fresh sample with this density gradient did not show a better resistance to freezing with this density gradient in comparison with the control sample.     

Alternative procedures for the cryopreservation of brown bear ejaculates depending on the flexibility of the “in cooling” period (5 °C)

The adaptability of cryopreservation protocols for brown bear spermatozoa collected under field conditions and frozen in a nearby laboratory (transported for a few hours) or shipped to a reference laboratory for sex sorting (transported for a few days) was evaluated. Forty-nine electroejaculates from 15 mature brown bears were extended to 100 × 10⁶ sperm/mL in a TES-Tris-Fructose based extender and cryopreserved (−20 °C/min to −100 °C and stored at −196 °C). After thawing, the quality of the seminal samples was assessed for total (TM), progressive (PM) motility and kinetic parameters – by CASA –, and viability (VIAB), viable and non-apoptotic status (YOPRO−), high membrane mitochondrial potential (MIT) and intact acrosomes (iACR) – by flow cytometry –. In Experiment 1, we assessed different storage times (0, 0.5, 1 – control –, 4–5, 7–8 and 11–12 h) at 5 °C from final dilution to freezing. After thawing, non-equilibrated samples (0 h) showed lower values of iACR, TM and PM. No significant differences were found for the different periods of equilibration tested. In Experiment 2, we evaluated three long-term storage times (24, 48 and 72 h) at 5 °C before freezing using storage for 1 h as control. The post-thawing quality of brown bear spermatozoa declined markedly after 48–72 h of pre-freezing. In conclusion, our findings suggest the possibility of extending the pre-freezing cooling period up to 24 h post-collection without freezing. This knowledge should enable the adaptation of the freezing protocols for when a special handling conditions are required such as the shipment of seminal samples to technological centers for the pre-freezing application of enhancer spermatic biotechnologies.     

Use of commercial extenders and alternatives to prevent sperm agglutination for cryopreservation of brown bear semen

The objective of this study was to evaluate different bovine and canine commercial semen extenders for cryopreservation of brown bear ejaculates and the effect of semen collection directly into extender on sperm agglutination. Semen samples were obtained by electroejaculation from 13 adult males. In experiment 1, eleven ejaculates from eight bears were used to evaluate Bioxcell and Andromed as extenders, whereas in experiment 2, nine ejaculates from six bears were used to evaluate Triladyl canine, CaniPro, and Extender 2 as extenders. An extender specifically developed for brown bears (Test-Tris-fructose-egg yolk-glycerol, TTF-ULE/bear) served as a control extender in both experiments. After thawing, total and progressive sperm motility and sperm viability were greater (P < 0.05) for TTF-ULE/bear and Andromed extenders than for Bioxcell in experiment 1 and greater (P < 0.05) for TTF-ULE/bear extender than for Triladyl Canine, CaniPro, and Extender 2 in experiment 2. In experiment 3, addition of handling extender (TTF-H) to the semen collection tube for eight ejaculates from seven bears resulted in less (P < 0.05) sperm agglutination in fresh samples (score 0.5 ± 0.2 vs. 1.8 ± 0.4 in diluted and control samples, respectively) with no effect on pre-freeze and post-thawing semen quality. In conclusion, TTF-ULE/bear is the most suitable extender for brown bear semen cryopreservation, but comparable results can be obtained with the commercial extender Andromed. In addition, collection of ejaculates directly in TTF-H extender decreases sperm agglutination in fresh samples.     

Brown bear sperm double freezing: Effect of elapsed time and use of PureSperm® gradient between freeze–thaw cycles

The use of sexed spermatozoa has great potential to captive population management in endangered wildlife. The problem is that the sex-sorting facility is a long distance from the semen collection place and to overcome this difficulty two freeze–thaw cycles may be necessary. In this study, effects of refreezing on brown bear electroejaculated spermatozoa were analyzed. We carried out two experiments: (1) to assess the effects of the two freezing–thawing cycles on sperm quality and to analyze three different elapsed times between freezing–thawing cycles (30, 90 and 180 min), and (2) to analyze the use of PureSperm between freezing–thawing cycles to select a more motile and viable sperm subpopulation which better survived first freezing. The motility, viability and undamaged acrosomes were significantly reduced after the second thawing respect to first thawing into each elapsed time group, but the elapsed times did not significantly affect the viability and acrosome status although motility was damaged. Our results with the PureSperm gradient showed higher values of viability in freezability of select sample (pellet) respect to the rest of the groups and it also showed a significant decrease in the number of acrosome damaged. In summary, the double freezing of bear semen selected by gradient centrifugation is qualitatively efficient, and thus could be useful to carry out a sex-sorting of frozen–thawed bear spermatozoa before to send the cryopreserved sample to a biobank. Given the low recovery of spermatozoa after applying a selection gradient, further studies will be needed to increase the recovery rate without damaging of the cell quality.     

黑龙江省熊类年龄构成、性比及繁殖情况的初步研究

熊类是重要的狩猎动物,肉、皮、骨、脂均可利用,熊掌和熊胆尤为名贵,因而具有很高的经济价值。 目前,国际上对于熊类生态学的研究日益深入。近二十年来多次举行国际熊类研究专题讨论会并出版了论文集。研究方向集中于熊类的群体生态,行为生态及种群的管理方面并正朝着集约管理和合理利用的方向发展。     

蜂桶寨自然保护区小熊猫对生境的选择

2002年4月至11月,在邛崃山系宝兴蜂桶寨自然保护区设点,应用资源选择函数和资源选择指数研究了小熊猫对生境的选择和利用。结果表明,影响小熊猫生境选择的关键因子是水源距离、竹子基径、灌木密度;次关键因子是坡位、树桩密度、倒木密度;次要因子是坡向、乔木密度;而郁闭度、坡度、植被类型、乔木高度、灌木高度、树桩高度、树洞密度、人为干扰的影响不明显。小熊猫喜欢在水源较近(<250m)、竹子长势良好(基径大于4 0mm)、灌木和乔木密度大(大于1 5株 20m2)、树桩和倒木数量多(>1根 400m2)、中上坡位、南坡的针叶林或针阔混交林活动。小熊猫的生境资源选择函数为:logit(P)=-13 527-3 180×水源距离 2 702×竹子基径 2 582×灌木密度 2 134×树桩密度 2 104×坡位 1 622×倒木密度-1 066×坡向 0 934×乔木密度……小熊猫对生境的选择概率为:P=elogit(P) (1 elogit(P))     

Selectivity of tree species as activity target of brown bear in taiga

Data on trees with signs of activity of brown bears (Ursus arctos L.) were collected in Yar raion of Udmurtia, in the Pechora-Ilych State Reserve (Komi Republic), and the Sayan-Shusha State Reserve (Western Sayan, Krasnoyarsk krai). The forest stands in all studied areas consist of deciduous and coniferous species. The percentage of deciduous trees varies from 10 to 76%, averaging 35%; conifers are represented by four or five species. The author has analyzed papers that describe more than fifty bear trees. The original data on the species composition of bear trees and the fraction of bear trees in the local forest stands was analyzed with Student’s t-test. It has been deduced that bears definitely prefer coniferous species, especially fir and spruce. Bears can also mark deciduous trees, but if conifers are present, they choose coniferous rather than deciduous species.     

The permeability of highway in Gorski kotar (Croatia) for large mammals

The highway from Zagreb to Rijeka stretches 68.5 km through a wildlife core area in Gorski kotar (Croatia). It has 43 viaducts and tunnels, and one specifically constructed (100 m wide) green bridge (Dedin). One quarter of the total highway length consists of possible crossing structures. At Dedin green bridge, a total of 12,519 crossings have been recorded during 793 different days of active infrared monitors being in operation, or 15.8 crossings per day. Two monitored tunnel overpasses had 11.2 and 37.0 crossings per day, respectively, whilst 4.3 crossings occurred per day under one monitored viaduct. Of those crossings, 83.2% were by ungulates and 14.6% by large carnivores. Radio-tracked large carnivores, brown bear (Ursus arctos), grey wolf (Canis lupus) and Eurasian lynx (Lynx lynx), expressed strong positive selection for tunnels and viaducts, whilst avoiding small underpasses or bridges. Selection for the use of Dedin green bridge was equal to its availability. We conclude that this green bridge, constructed as a measure to mitigate the negative effects of the studied highway, served its purpose acceptably. Territorial and dispersing radio-tracked large carnivores crossed the highway 41 times, using both sides of the highway as parts of their home ranges. Overall, the highway in Gorski kotar does not seem to be a barrier. This demonstrates that it is possible to maintain habitat connectivity during the process of planning the highway route.     

莫莫格自然保护区白鹤秋季迁徙停歇期觅食生境选择

2008年秋季(10月8日—20日)及2009年秋季(9月20日—10月16日),通过样方法对觅食生境11个生态因子进行调查,利用卡方检验、资源选择指数和资源选择函数在莫莫格保护区对秋季迁徙停歇期白鹤觅食生境选择进行研究。结果表明,白鹤对距人为干扰源距离、植被密度、盖度、高度、植物性食物密度以及水深均具有选择性,但对宏观尺度干扰因子的选择性较低。其偏好觅食生境的特点为:距一级路>5000m,>二级路1500m以上,>三级路1000m以上,>居民点1000m以上,农田>1000m;植被密度20~50株/m2,盖度<10%,高度<20cm,扁杆藨草密度1~50株/m2,藨草密度1~10株/m2,水深40~60cm。白鹤秋季觅食生境资源选择函数为Logistic(P)=0.663+0.565×与一级道路距离+0.042×与二级道路距离+0.519×与三级道路距离+0.353×与居民点距离+0.169×与农田距离–0.455×植被密度–0.618×植被盖度–0.548×植被高度–0.158×扁杆藨草密度–0.404×藨草密度+0.920×水深,T(x)=eLogistic(p)/[1+eLogistic(p)],模型正确预测率为82.9%。     

甘肃祁连山脉雪豹及其同域分布大型食肉动物时间生态位关系

<正>群落内多物种如何共存是群落生态学和生物多样性研究的核心内容之一。经典物种共存理论强调物种之间的生态位分化,侧重于物种对环境的需求,Hutchinson (1957)提出超体积生态位概念,认为物种适合度是由多个因素共同决定,即物种只有在满足其生态位需求的多维空间,