脊髓灰质炎病毒感染赤麂细胞株的实验研究

脊髓灰质炎病毒的敏感细胞范围一直局限于灵长类动物细胞。用该病毒感染处理非灵长类的赤麂肺二倍体细胞(KIZ-7901),可诱导9.0—14.7%的细胞出现染色体畸变,除见有染色体或单体断裂外,还见有断片、双着丝粒、环、互换、易位和缺失等。也可引致姐妹染色单体交换率(SCE)的显著提高。脊髓灰质炎病毒感染赤麂细胞后,不产生特异的CPE,不明显影响细胞群体的增殖。应用病毒滴定、包涵体检查,间接免疫荧光试验和电镜检查,都可查见细胞内有病毒或其抗原物质存在。这提示脊髓灰质炎病毒在非灵长类的赤麂细胞内呈现非杀细胞性感染。     

同域分布的黑麂和小麂的时空生态位分化

生态位分化是解释物种共存的重要理论。Schonenr认为空间维度对生态位差异形成的影响程度最高,营养维度次之,时间维度则最后被启动。为验证这一假说,我们于2018年12月至2019年11月,在浙江清凉峰国家级自然保护区龙塘山区域采用公里网格法布设52台红外相机,对同域分布且食性相似的黑麂和小麂种群进行监测,计算平均拍摄率,分析不同季节黑麂和小麂对不同植被类型、海拔、坡位、水源距离的选择差异,并运用核密度估计法分析二者日活动节律及重叠程度。结果显示,(1) 黑麂和小麂对空间生态位的选择出现分化：黑麂偏好针阔混交林,主要栖息于1 301 ~ 1 500 m的高海拔地区,回避下坡位和谷地,在距离水源较近的区域内活动频繁;小麂则偏好落叶阔叶林,主要栖息于901 ~ 1 100 m的中海拔地区,偏好中坡位,回避谷地,对水源距离没有明显的选择倾向。(2) 黑麂和小麂均为晨昏性活动的昼行性动物,二者全年日活动节律重叠程度较高 (Δ4 = 0.86),仅冬季较低 (Δ1 = 0.65)。上述结果支持Schonenr的假说：首先,空间维度对物种生态位分化的影响大于时间维度,龙塘山区域黑麂和小麂主要通过选择不同的栖息地来增加空间回避,降低二者的竞争;其次,这种模式存在季节差异,冬季由于可利用的资源减少,竞争加剧,二者通过调整日活动节律,增加时间生态位分化程度,从而实现同域共存。     

黑麂肝、小肠和大肠的组织学结构及Ghrelin的分布

研究了成年雌性黑麂(Muntiacus crinifrons)的肝、小肠和大肠的组织学结构及Ghrelin的分布。采用H.E染色法观察组织学结构,免疫组化PV-9000两步法并结合DAB显色技术确定Ghrelin的分布。结果表明,黑麂的肝组织分为被膜、肝小叶、肝中央静脉、门管区等结构。被膜为浆膜结构,肝小叶不明显。肝细胞以中央静脉为中心,呈放射状排列。肝血窦的形状不规则。肠黏膜上皮为单层柱状上皮,小肠、盲肠和结肠的黏膜肌层很薄,管壁皱襞与肠绒毛等形态在消化道各部也存在差异。免疫组化结果显示肝细胞中有Ghrelin阳性细胞的表达;在肠道,免疫阳性细胞在十二指肠、空肠、回肠、盲肠和直肠的黏膜、黏膜下层和肌层均有分布,尤其在肠绒毛上皮和黏膜下层分布较多。黑麂肝、小肠和大肠结构与哺乳动物基本相似,但无十二指肠腺;Ghrelin阳性细胞在肝、小肠和大肠均有分布,这表明Ghrelin可能对消化有一定的调节作用。     

Isolation and characterization of microsatellite markers in black muntjac (Muntiacus crinifrons)

The black muntjac (Muntiacus crinifrons) is a vulnerable species found in the mountains of eastern China, about which little is known. Here we develop 11 polymorphic microsatellite loci from a (CA)(n) -enrichment library for the animal. In the analyses of 25 individuals sampled, unbiased expected heterozygosity levels varied from 0.686 to 0.876 and the number of alleles per locus varied from five to nine. Results that 11 microsatellite loci were highly polymorphic indicated that these markers are sufficiently powerful to address such questions as parentage, mating system and population genetic structure of M. crinifrons.     

黑麂线粒体基因组序列分析

采用PCR产物直接测序方法测定了黑麂线粒体基因组全序列 ,初步分析了其基因组特点并定位了各基因的位置 .结果显示 :黑麂的线粒体基因组全序列长度为 1 6 35 7bp ,可编码 2 2种tRNA、2种rRNA、1 3种蛋白质 ,碱基组成及基因位置与小麂、赤麂和其它哺乳类动物的线粒体基因组相似 ;模拟电子酶切图谱与先前的报道基本一致 ;基于细胞色素b的全基因序列 ,分别以最大简约法、N J法、最大似然数法与其它 1 4种鹿类动物的相应序列进行了聚类分析 ,构建出相似的系统进化树 :初步确定了麂亚科动物在鹿科中处于与鹿亚科、北美鹿亚科并列的进化地位 .在此基础上 ,进一步以黑麂、赤麂、小麂的线粒体编码RNA和编码蛋白质的基因序列构建系统进化树 ,分析了三者的亲缘关系 .结果表明 :黑麂和赤麂亲缘关系较近 ,是较新的物种 ,而小麂是较为原始的物种     

毛冠鹿与3种麂属动物的线粒体细胞色素b 的系统进化分析

采用PCR直接测序法,获得毛冠鹿、小鹿、赤麂和黑麂等4种麂亚科动物线粒体细胞色素b核酸序列366bp,麂属动物之间的序列差异为3.5%-4.6%,毛冠鹿属与麂属之间的序列差异为9.29%-10.11%,麂属3种动物分歧时间约为142-184万年,毛冠鹿与麂属动物的分歧时间约为370万年。结合鹿科其余3亚科动物的同源序列,用MEGA软件的Neighbor-Joining Method(NJ法）和最大简约法构建系统进化树。结果表明,小麂、赤麂与黑麂组成一个单系群,毛冠鹿细胞色素b核苷酸序列与麂属的分化程度似已达到属间水平。     

浙江黑麂栖息地评价及保护对策

运用地理信息系统技术,在分析黑麂栖息地的地形、植被、水系和人为干扰等地理特征的基础上,系统研究九龙山和古田山自然保护区黑麂栖息地分布、栖息地质量与空间格局。结果表明,在九龙山自然保护区有黑麂潜在栖息地3172．6hm^2,由于人类活动的影响导致栖息地丧失501．5hm^2,目前尚存的黑麂适宜栖息地2671．1hm^2。它们主要分布于上寮坑、九龙山、内北坪、外九龙和大岩前附近区域。古田山自然保护区有黑麂潜在栖息地4635．75hm^2,在人类活动影响下丧失栖息地1118．11hm^2,目前尚存的黑麂适宜栖息地3517．64hm^2。它们主要分布于青尖、古田山、巧观尖、催顶尖附近区域。目前九龙山和古田山自然保护区内的黑麂栖息地基本上处于多斑块破碎化状态。为了使黑麂能得到更好的保护,对这两个研究区域黑麂栖息地的恢复和重建工作亟待加强。     

Evidence of higher levels of testosterone during the velvet period in muntjac than in other cervids

Previous studies have shown that despite having a clear seasonal fluctuation in fecal testosterone concentration, the significantly lower testosterone levels found in velvet stags of the nonseasonal breeder muntjac (Muntiacus sp.) apparently did not stop their spermatogenesis as in other deer species. In the present study, in vitro cultivated Leydig cells isolated from adult stags of three native deer species of Taiwan were treated with androstenedione, with or without adding human chorionic gonadotropin. Results showed that, unlike the two seasonal breeders, sika deer (Cervus nippon) and sambar deer (Rusa unicolor), Leydig cells of velvet muntjac had no dramatic reduction in or even maintained the full capability of their testosterone productivity compared with the hard-antlered stage. The decrease in fecal testosterone level observed earlier in muntjac during the velvet period was probably due to a reduction of number of Leydig cells. These results support the hypothesis that testosterone production in muntjac during its velvet period might never be low enough to trigger the quiescent phase of the reproduction cycle.     

妊娠黑麂（Muntiacus crinifrons）大体解剖分析

为黑麂的保护和人工繁殖提供生物学资料,对一只因盗猎致死的妊娠黑麂进行了解剖。其特点是：上腭有15排腭褶;上颌右侧犬齿脱落,前臼齿和臼齿的齿冠磨损较严重,齿式为0·1·3·3/3·1·3·3=34;瘤胃、网胃、瓣胃、皱胃容积分别为71.6%、9.4%、11.4%、7.6%;小肠7248 mm、大肠5269 mm,分别是体长的6.9倍和5.0倍;肝3叶,重598.5 g,无胆囊;左肺3叶,右肺4叶,重474.4 g;心脏重184 g,占身体总重量的0.8%;双角子宫,胚胎重98.5 g,长14.8 mm,胚体弯曲呈＂C＂形,眼泡明显,鼻窝、耳泡、鳃弓、肢芽出现。     

Spatial dynamics of Chinese Muntjac related to past and future climate fluctuations

Climate fluctuations in the past and in the future are likely to result in population expansions, shifts, or the contraction of the ecological niche of many species, and potentially leading to the changes in their geographical distributions. Prediction of suitable habitats has been developed as a useful tool for the assessment of habitat suitability and resource conservation to protect wildlife. Here, we model the ancestral demographic history of the extant modern Chinese Muntjac Muntiacus reevesi populations using approximate Bayesian computation (ABC) and used the maximum entropy model to simulate the past and predict the future spatial dynamics of the species under climate oscillations. Our results indicated that the suitable habitats for the M. reevesi shifted to the Southeast and contracted during the Last Glacial Maximum, whereas they covered a broader and more northern position in the Middle Holocene. The ABC analyses revealed that the modern M. reevesi populations diverged in the Middle Holocene coinciding with the significant contraction of the highly suitable habitat areas. Furthermore, our predictions suggest that the potentially suitable environment distribution for the species will expand under all future climate scenarios. These results indicated that the M. reevesi diverged in the recent time after the glacial period and simultaneously as its habitat’s expanded in the Middle Holocene. Furthermore, the past and future climate fluctuation triggered the change of Chinese muntjac spatial distribution, which has great influence on the Chinese muntjac’s population demographic history.