Detection of parasitizing coccidia and determination of host crane species, sex and genotype by faecal DNA analysis

In Japan, the three main crane species are the endangered red-crowned crane (Grus japonensis) inhabiting Hokkaido, the northernmost island of Japan; the vulnerable hooded crane (Grus monacha); and the vulnerable white-naped crane (Grus vipio). Both the hooded and white-naped cranes migrate in winter to Izumi in Kyushu, the southern island of Japan. In this study, we investigated the cranes and their coccidian parasites, through a targeted molecular approach using faecal DNA to develop a noninvasive method for infectious disease research. To determine the origin of noninvasively collected faecal samples, host species were identified by sequencing a region of approximately 470 bp of the mitochondrial 16S ribosomal RNA gene in the faecal DNA. Furthermore, to avoid sample redundancy, individual determination was performed by fragment analysis using microsatellite and sex-linked markers. For microsatellite genotyping, previously reported markers and markers isolated in this study were examined, and seven loci for red-crowned cranes, eight for hooded cranes and six for white-naped cranes displayed polymorphisms. A low error rate was demonstrated by comparing microsatellite data generated from faecal DNA samples with that generated from feather DNA samples, indicating a high reliability. Polymerase chain reaction-based capillary electrophoresis (PCR-CE), employing genetic markers in the second internal transcribed spacer (ITS2) of nuclear ribosomal DNA, was employed to detect crane coccidia. The sensitivity of detection of PCR-CE using faecal DNA was inferior to that with traditional microscopy; however, our results suggest that PCR-CE can depict crane coccidia diversity with higher resolution and it is a useful tool to characterize community composition of coccidia in detail.     

Allozyme evidence for crane systematics and polymorphisms within populations of Sandhill, Sarus, Siberian, and whooping cranes.

Electrophoretic analysis of proteins yielded evidence on the relationships of species of cranes and on genetic diversity within populations of some species. Diversity within the Greater Sandhill crane and a Florida population of the Florida Sandhill crane was similar to that of most other vertebrates, but diversity was low in the Mississippi Sandhill crane, in the Okefenokee population of the Florida Sandhill crane, and within the Siberian and Sarus cranes. Diversity was surprisingly high among whooping cranes, whose number dropped to less than 25 early in this century. Phylogenetic analysis, using both character state and distance algorithms, yielded highly concordant trees for the 15 species. The African crowned cranes (Balearica) were widely divergent from all other cranes. Species of Anthropoides, Bugeranus, and Grus clustered closely but sorted into two lineages: a Whooper Group consisted of the whooping, common, hooded, black-necked, white-naped, and red-crowned cranes of genus Grus; and a Sandhill Group included the Sandhill, Siberian, Sarus, and Brolga cranes of genus Grus, the wattled crane of genus Bugeranus, and the Demoiselle and blue cranes of genus Anthropoides.     

圈养丹顶鹤与白枕鹤杂交繁育

我园一只雄性丹顶鹤与一只雌性白枕鹤于2001年开始自然配对产蛋,3年共产51枚,经人工孵化育雏,目前存活15只。杂种鹤在第4周后进入快速增长阶段,在12周达到最高峰,16周后增长速度明显下降,18周后部分指标停止增长。外观上,出生至3月龄时,幼鹤整体颜色都呈黄褐色,以后变化基本一致。目前,2001及2002年存活杂种鹤颜色以白色为主,夹杂着灰色羽毛。     

Coccidia of whooping cranes

Coccidial oocysts were observed in 6 of 19 fecal samples from free-ranging whooping cranes (Grus americana) and 4 of 16 samples from captive whooping cranes. Eimeria gruis occurred in four free-ranging whooping cranes and E. reichenowi in two free-ranging and two captive whooping cranes. Fecal samples from two captive cranes contained oocysts of Isospora lacazei which was considered a spurious parasite. Oocysts of both species of Eimeria were prevalent in fecal samples collected from three free-ranging Canadian sandhill cranes (G. canadensis rowani) from whooping crane wintering grounds in Texas. These coccidia were prevalent also in fecal samples from 14 sandhill cranes (of 4 subspecies) maintained in captivity at the Patuxent Wildlife Research Center in Maryland.     

Effect of heterospecifics on foraging of endangered red-crowned and white-naped cranes in the Korean Demilitarized Zone

We determined how the presence of heterospecific individuals in the vicinity of a focal individual affects the behavior of two critically endangered species of cranes on their wintering grounds at Cheolwon in the Korean Demilitarized Zone. The red-crowned crane, Grus japonensis, is larger than the white-naped crane, Grus vipio, and it dominates the white-naped crane in aggressive interactions. We showed that the dominant species increases foraging activity in the presence of the subordinate species presumably because of scrounging of food from the subordinate. Because interspecific interactions may affect avian endangerments these behavioral findings should be taken into account when managing winter refuges for the two endangered crane species.     

Pulmonary lesions in disseminated visceral coccidiosis of sandhill and whooping cranes

Fifty cranes, consisting of 46 sandhill (Grus canadensis) and four whooping cranes (Grus americana), were studied. Eighteen sandhill cranes and the four whooping cranes were naturally infected with disseminated visceral coccidiosis (DVC). The remaining sandhill cranes were chicks experimentally infected with oocysts of Eimeria reichenowi and/or E. gruis; five chicks served as controls. There were no clinical signs attributed to respiratory infection. Necropsy of naturally infected adult birds revealed nodules in many organs, including the lung, air sacs, trachea and nares. Artificially infected sandhill cranes and the whooping crane chicks that died from DVC had congestion and consolidated areas in the lung with frothy fluid in the airways. Grossly visible nodules were observed from 10 days postinoculation. Granulomatous pneumonia and tracheitis were observed with light microscopy. Lesions were associated with merogonic and gametogonic stages of eimerian coccidia. Granulomas and granulomatous foci contained parasitized large mononuclear cells. Merogonic stages were seen in lymphoid cells by ultrastructural examination. Oocysts were observed in the trachea and bronchial mucosa and admixed with exudate in the airways, indicating that crane eimerians can complete their life cycle at these sites. Of the few eimeriid coccidia that have extraintestinal stages of development in birds and mammals, only the species in cranes complete their life cycle in both the digestive and respiratory tracts.     

Coccidiosis of sandhill cranes (Grus canadensis) wintering in New Mexico

The feces of 212 sandhill cranes (Grus canadensis) collected in central New Mexico from October 1982 to January 1983 and from October 1983 to January 1984 were examined to determine the prevalence of coccidial oocysts. One hundred forty-five granulomatous nodules from the viscera of 64 cranes and samples of lung, small intestine, and large intestine from 58 birds were examined by light and transmission electron microscopy for the presence of intestinal or extraintestinal coccidiosis. Of the 212 fecal samples, 160 (75%) were positive for oocysts of Eimeria, including E. gruis in 139 (66%) and E. reichenowi in 118 (56%) of the samples. Eimeria bosquei sp. n. was found in two (approximately 1%) of the fecal samples. Subspheroid to ovoid oocysts of this new species are 19-27 X 14-19 (23.6 X 17.1) micrograms with ovoid sporocysts 10-14 X 7-11 (12.3 X 9.3) micrograms. A rough, heavily pitted outer oocyst wall, sporocyst residuum, Stieda and substieda bodies, and multiple polar bodies are present. The polar bodies, of varying sizes, always aggregate at the apex of the sporulated oocyst. An Adelina sp. was found in one (0.5%) crane. Coccidian developmental stages were found in the epithelium and lamina propria of the small and large intestine. Disseminated granulomatous nodules were found in the oral mucosa, esophagus, heart, descending aorta, liver, small intestine, mesenteries, and parietal peritoneum. Unique cell types resembling coccidian asexual and sexual stages were observed by light and electron microscopy in some of the nodules.     

基于极坐标定位的丹顶鹤多尺度越冬生境选择研究——以江苏盐城自然保护区为例

丹顶鹤作为湿地环境的指示物种,其种群数量逐年下降引起了野生动物保护工作者的广泛关注。近年来,随着研究的深入,鸟类生境选择具有多尺度特征的观点逐渐成为研究者的共识。江苏盐城自然保护区是丹顶鹤大陆西线种群最重要的越冬栖息地,选择该区域进行多尺度越冬生境选择研究对于丹顶鹤保护具有重要的现实意义。基于遥感影像和丹顶鹤越冬分布的调查,本文用极坐标的方法在GIS软件中对丹顶鹤分布定位进行制图。借助Arc GIS 10.2和fragstats 4.2软件,从点缓冲、景观和区域3个尺度系统分析丹顶鹤越冬生境多尺度利用特征。在点缓冲尺度上,通过缓冲分析和叠置分析描述丹顶鹤生境利用的强度,发现丹顶鹤连续利用生境和经常利用生境的植被类型是碱蓬,偶尔在芦苇中活动,偏好生境区植被覆盖度在30%以下,植被高度在60 cm以下。在景观尺度上,丹顶鹤偏好程度较高的前三类生境为芦苇沼泽、碱蓬沼泽、明水,对互花米草沼泽具有负向选择。在区域尺度上,受道路和土地利用干扰影响,丹顶鹤主要分布于保护区核心区的低干扰区域;除了干扰之外还与生境面积、生境破碎程度密切相关。综合三个尺度,研究得出丹顶鹤生境选择具有明显的多尺度利用特征,丹顶鹤保护需要考虑多尺度生境保护问题。     

黑龙江省乌裕尔河流域丹顶鹤等珍稀涉禽航空调查报告

利用Y-11型单翼轻型飞机,航高100米,航速140公里/小时,采用且测全区调查方法,对黑龙江省乌裕尔河流域丹顶鹤等涉禽进行了航空调查,续航里程2184公里,调查面积共计为209,000公顷。结果发现调查区内有丹顶鹤(Orus japoncnsis)173只,白枕鹤(Grus vipio)19只,白鹤(Grus leucogeraous)24只,白鹳(Ciconia ciconia)19只等。     

辽宁双台河口自然保护区丹顶鹤繁殖生境变化及其繁殖容量分析

运用面积转换矩阵和景观指数对双台河口自然保护区丹顶鹤繁殖生境的变化进行分析。结果表明 ,该地区丹顶鹤繁殖生境的面积不断减小 ,景观异质性和破碎化程度不断增强 ,生境质量不断下降。生境质量的变化将直接引起在该地区丹顶鹤繁殖数量的变化 ,从 1989～1998年 ,在双台河口自然保护区繁殖的丹顶鹤数量逐年下降 ,丹顶鹤的繁殖容量日益减小 ,原因是道路和油井开发强度的不断增大等人类活动的影响。     

苏北地区丹顶鹤越冬种群数量及栖息地分布动态变化

利用资料追踪和历史时期土地利用变化数据深入研究了有资料记载以来苏北地区丹顶鹤越冬种群数量变化与栖息地分布的时空响应。结果显示,20世纪80年代之初,丹顶鹤越冬种群广泛分布于苏北地区,尤其是长江中下游的内陆湖泊、沼泽及江苏沿海滩涂湿地;20世纪90年代之后丹顶鹤分布区域逐渐向沿海滩涂湿地转移,主要分布于盐城市的射阳县境内;进入21世纪,丹顶鹤分布区域集中于盐城国家级珍禽保护区核心区。与此同时,丹顶鹤越冬种群数量也经历了十分明显的变化过程,与20世纪90年代相比,数量减少50%以上。丹顶鹤越冬种群数量及栖息地分布变化与栖息地分布点周围的土地利用方式及人为干扰有关,栖息地面积丧失和景观破碎化是丹顶鹤越冬种群数量减少的主要原因。     

Cryopreservation effects on sperm function and fertility in two threatened crane species

The capacity to cryopreserve semen from captive cranes facilitates production of offspring from behaviorally incompatible or geographically separated pairs, and allows for long-term preservation of valuable genetic materials. The present study sought to develop effective cryopreservation protocols for whooping (Grus americana) and white-naped (Grus vipio) cranes, through examining the influences of two permeating (DMA and Me₂SO) and one non-permeating (sucrose) cryoprotectants, as well as vitamin E on post-thaw sperm survival. In Study 1, ejaculates (whooping: n = 10, white-naped: n = 8) were collected and cryopreserved in one of six cryo-diluents (crane extender with: DMA; DMA+0.1M sucrose; Me₂SO; Me₂SO+0.1M sucrose; 0.1M sucrose; 0.2M sucrose) using a two-step cooling method. Frozen samples were thawed and assessed for overall motility, motion characteristics, membrane integrity, morphology, and ability to bind to the inner perivitelline membrane (IPVM). In Study 2, whooping crane ejaculates (n = 17) were frozen in crane extender containing Me₂SO alone or with vitamin E (5 μg/mL or 10 μg/mL). Frozen samples were thawed and assessed as in Study 1, except the binding assay. White-naped crane sperm were more tolerant to cryopreservation than whooping crane (15% vs 6% post-thawed motility). In both species, sperm cryopreserved in medium containing Me₂SO alone displayed higher post thaw survival and ability to bind to IPVM than the other cryodiluent treatments. Vitamin E supplementation exerted no benefits to post thaw motility or membrane integrity. The findings demonstrated that there was species specificity in the susceptibility to cryopreservation. Nevertheless, Me₂SO was a preferred cryoprotectant for sperm from both whooping and white-naped cranes.     

Individual (non-social) behavioral acts of hooded cranes<Emphasis Type="Italic"> Grus monacha</Emphasis> wintering in Izumi,Japan

The purpose of this paper is to make a fundamental tool (an ethogram) to help understand the life of hooded cranes Grus monacha and also the phylogenetic relationships among congeneric crane species from the ethological viewpoint. Individual behavioral acts were enumerated with quantitative attributes depending on the analysis of video images recorded in Izumi, wintering grounds of the species in Japan, during 1995–1999. Several new ethons and activities of individual behavior are cited here, such as Soak, Wing-droop and Bill-turn of Care of the body surface, Ruffle-bow-up, High-step-stretch and Foot-dangle of Comfort movements, Walk-back, Trot, three modes of Alight and Dangle-leg-glide of Locomotory, and Nibble-search and Carry of Foraging. These ethons have so far rarely been mentioned in inventories of individual behavior on cranes. It is necessary to obtain not just qualitative but quantitative data of individual acts of other cranes for the comparison of such closely related species as Eugrus group members.     

扎龙保护区春季丹顶鹤觅食的农田生境利用分析

为了更深入地了解候鸟丹顶鹤春季利用的农田生境情况,以期为后续的该时期鹤类监测及觅食农田而产生的补偿评估奠定基础并制定相应的管理策略,2002—2015年的5月采用定点观察法、二维坐标法、样方法、GPS定位和因子分析等方法,对扎龙保护区丹顶鹤春季利用的农田生境特征进行了调查。分析结果表明:(1)春季,丹顶鹤觅食偏好选择的农作物为玉米(100%),偏好选择2—5片叶(94.32%)、回避1片叶和超过6片叶(100%)的玉米苗;(2)丹顶鹤觅食利用的农田生境通过有效食物因子和干扰因子进行判定;(3)有效食物因子包括作物高度、作物密度、作物面积、距明水面距离、距芦苇沼泽距离等;其中,选择的作物密度约为8—10株/m~2、作物高度为6—7cm,作物面积4—50hm~2大小不等、距明水面和芦苇沼泽距离相似约为0.4km左右;(4)干扰因子包括季节性弱干扰因子如道路等和强干扰因子如居民区等,针对弱干扰因子丹顶鹤采取不排斥的忽视方式,约保持在0.20km以上;针对强干扰因子丹顶鹤采取远离的方式进行回避,保持在1.20km以上。基于上述分析,建议扎龙保护区在春季巡护时应对人为活动区和农田给予更多的关注。     

Obtaining an Interspecific Hybrid of Cranes by Artificial Insemination with Frozen–thawed Semen

Studies of artificial insemination of cranes and cryoconservation of their semen have been carried out in the nursery of rare species at the Oka Biosphere Reserve for many years. The criterion of successful cryoconservation of the semen is the obtaining of fertilized eggs after artificial insemination by the thawed semen. An experiment is described on artificial insemination of females of the white-naped crane Grus vipio by the frozen–thawed semen of the Siberian white crane G. leucogeranus after one-year storage of semen in liquid nitrogen. As a result, an interspecific hybrid of cranes was obtained, which confirmed the possibility of producing a bank of cryoconserved crane semen. The use of the white-naped crane females was due to the absence of conspecific males and unavailability of Siberian white crane females. Problems of artificial insemination and cryoconservation of semen of rare crane species are discussed.     

Obtaining an Interspecific Hybrid of Cranes by Artificial Insemination with Frozen–thawed Semen

Studies of artificial insemination of cranes and cryoconservation of their semen have been carried out in the nursery of rare species at the Oka Biosphere Reserve for many years. The criterion of successful cryoconservation of the semen is the obtaining of fertilized eggs after artificial insemination by the thawed semen. An experiment is described on artificial insemination of females of the white-naped crane Grus vipio by the frozen–thawed semen of the Siberian white crane G. leucogeranusafter one-year storage of semen in liquid nitrogen. As a result, an interspecific hybrid of cranes was obtained, which confirmed the possibility of producing a bank of cryoconserved crane semen. The use of the white-naped crane females was due to the absence of conspecific males and unavailability of Siberian white crane females. Problems of artificial insemination and cryoconservation of semen of rare crane species are discussed.     

丹顶鹤春迁期觅食栖息地多尺度选择——以双台河口保护区为例

双台河口保护区是世界濒危鸟类丹顶鹤大陆种群西线群体不同生活史阶段的重要栖息地。2008-2010年3-4月,采用定点观察法、二维坐标法、方差分析、因子分析等方法对保护区内春迁期丹顶鹤觅食栖息地的多尺度选择进行了监测分析。研究结果表明：(1)春迁期,该保护区丹顶鹤觅食栖息地选择包括2个尺度3个选择,即大生境尺度内觅食生境类型选择和小生境尺度内觅食区选择、觅食微生境选择;(2)觅食生境类型偏好选择芦苇沼泽(90.00%),也偶选玉米地、泥滩、草甸等生境;(3)觅食区选择通过宏生境因子和干扰因子来判定,宏生境因子包括明水面、堤坝和火烧地,距离均在30m以内;干扰因子包括居民区和油田等强干扰因子和道路等弱干扰因子,均采取远离的方式进行回避,居民区保持在1km以上,油田保持在2km以上,道路保持在300-500m;(4)觅食微生境选择通过微生境因子来判定,包括植被高度、植被密度、植被直径等,选择盖度小(<5%)且植被高度小于喙长的区域觅食。     

Dietary Exposure of the Red-Crowned Crane (Grus japonensis) to Total and Methyl Mercury in Zhalong Wetland,Northeastern China

To determine the dietary exposure of the migratory red-crowned crane to mercury (Hg), this study analyzed the concentrations of total mercury (T-Hg) and methyl mercury (MeHg) in its prey, i.e., reeds and three aquatic animal families (Perccottus glenni Dybowski, Cybister japonicus Sharp, and Viviparidae) in northeastern China. Results indicated that the Hg concentration in Zhalong Wetland was elevated through the food chain, and the prey of the red-crowned crane contained measurable levels of T-Hg and MeHg. In prey tissues, MeHg was the main form of the Hg element and accounted for 61 % of total Hg concentration in Viviparidae, 58 % in C. japonicus Sharp, and 85 % in P. glenni Dybowski. The highest T-Hg and MeHg concentrations ranged from 1.66 to 3.89 ppm and from 1.12 to 2.67 ppm, respectively, and they were detected in the feathers of the red-crowned cranes. The lowest T-Hg concentration was determined in the excretions of wild red-crowned cranes at 0.21 ppm; furthermore, the content of MeHg was below the detection limit. In Zhalong Wetland, the level of dietary exposure of the population of red-crowned cranes to Hg is below the threshold of Hg toxicity. Moreover, eggshells are suitable indicators of Hg risk levels to the red-crowned crane.     

黑龙江扎龙自然保护区白枕鹤巢址选择特征

为了探讨白枕鹤Grusvipio的巢址选择特征,2007～2009年的4～5月,在黑龙江省扎龙自然保护区采用定点观察法、样方调查法、因子调查法和因子分析及显著性检验等,对白枕鹤的营巢生境类型和营巢生境特征进行了调查。结果表明,白枕鹤对营巢生境类型的要求比较单一,仅选择芦苇沼泽;巢址生境特征具有明显的偏好,即(1)距干扰区的距离多在1.5km以上(86.36%),(2)巢周的水深多小于16cm(95.45%),(3)巢周明水面面积多小于200m2(90.90%),(4)巢距火烧地的距离以1.0～2.5km居多(63.64%),(5)剩余苇丛面积多小于200m2(86.37%)、距剩余苇丛距离多大于30m(81.82%)、剩余苇丛密度多小于500株/m2(77.27%)、剩余苇丛高度以大于1.5m的居多(72.73%)。进一步分析表明,白枕鹤对营巢生境特征的要求具有普遍性和特殊性,表现出一定的适应能力,火烧地、剩余苇丛、干扰区和水是其营巢的主要环境特征。     

气候变化对丹顶鹤秋季迁徙路线潜在生境适宜性的影响

目前全球变暖趋势的加剧对丹顶鹤等大型濒危水禽的栖息地造成了严重的威胁。由于监测方法和技术手段的限制,丹顶鹤在迁徙路线上潜在生境的分布范围尚不清楚,气候变化对丹顶鹤迁徙路线生境适宜性的影响机理有待进一步研究。基于138个丹顶鹤样本分布信息和19种环境变量数据,利用 BIOMOD2 软件包构建了丹顶鹤潜在生境评价的组合模型,对丹顶鹤在亚洲东部秋季迁徙路线上的生境适宜性进行数值模拟,并预测SSP1.2-6气候背景下2021-2040年、2041-2060年、2061-2080年、2081-2100年四个不同阶段的丹顶鹤潜在生境范围的变化趋势。研究结果表明:与单模型的模拟结果相比,集成9种单模型的BIOMOD2组合模型预测精度更高。集成模型的重要性分析表明,气温日较差是丹顶鹤生境适宜性变化的最重要的影响因子。受气候变化的影响2021-2040年、2041-2060年、2061-2080年、2081-2100年丹顶鹤潜在生境的面积将分别减少到2.60×10⁵km²、2.58×10⁵km²、2.75×10⁵km²、2.56×10⁵km²,迁徙路线上胶东半岛和环渤海地区适栖生境面积减少的最为显著。本研究对于迁徙路线上珍稀水禽潜在适宜生境的模拟及全球变化背景下珍稀水禽栖息地的保育和修复具有重要意义。