Ex situ conservation status of an endangered Yangtze finless porpoise population (Neophocaena phocaenoides asiaeorientalis) as measured from microsatellites and mtDNA diversity

The Yangtze finless porpoise, Neophocaena phocaenoides asiaeorientalis, is all endangered small cetacean that occurs only in the middle and lower reaches of the Yangtze River of China. The establishment of a breeding population of the porpoise in Tian-e-Zhou Baiji National Natural Reserve represents the first attempt at ex situ conservation efforts for a cetacean species. With the goal of effective protection, management, and monitoring of this preserved population, we examined its genetic diversity using 930 bp of mtDNA control region sequences and 13 polymorphic microsatellite loci. A very low level of genetic variation (h = 0.6010 +/- 0.0029 s.d.; pi = 0.0007 +/- 0.0000002 s.d.) in the mtDNA control region sequences and a moderate genetic diversity (Ho = 0.5740 +/- 0.2575 s.d.) in the microsatellites were detected in the population. It is necessary to introduce more individuals with representative genetic variations into the reserve ill order to foml a larger and healthier group structure for long-term survival of the population. Successful establishment of the Yangtze finless porpoise population in the Reserve also provides a useful model for an ex situ conservation programme for other rare and endangered species. (c) 2005 International Council for the Exploration ofthe Sea. Published by Elsevier Ltd. All rights reserved.

Population status, threats and conservation of the Yangtze finless porpoise

The Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis) is currently limited to the middle and lower reaches of the Yangtze River from Yichang to Shanghai, China, and the adjoining Poyang and Dongting Lakes. Its population size has decreased remarkably during the last several decades due to the heavy impact of human activities, including overfishing of prey species, water development projects that cause attendant habitat loss and degradation, water pollution, and accidental deaths caused by harmful fishing gear and collisions with motorized vessels. It was estimated that the number of remaining individuals was down to approximately 1800 in 2006, a number that is decreasing at a rate as high as 5% per year. Three conservation measures - in situ and ex situ conservation and captive breeding have been applied to the protection of this unique porpoise since the early 1990s. Seven natural and two "semi-natural" reserves have so far been established. Since 1996, a small group of finless porpoises has been successfully reared in a facility at the Institute of Hydrobiology of the Chinese Academy of Sciences; three babies were born in captivity on July 5, 2005, June 2, 2007 and July 5, 2008. These are the first freshwater cetaceans ever born in captivity in the world. Several groups of these porpoises caught in the main stream of the Yangtze River, or rescued, have been introduced into the Tian'e-Zhou Semi-natural Reserve since 1990. These efforts have proven that, not only can these animals survive in the area, they are also to reproduce naturally and successfully. More than 30 calves had been born in the reserve since then, with one to three born each year. Taking deaths and transfers into account, there were approximately 30 individuals living in the reserve as of the end of 2007. Among eight mature females captured in April 2008, five were confirmed pregnant. This effort represents the first successful attempt at off-site protection of a cetacean species in the world, and establishes a solid base for conservation of the Yangtze finless porpoise. A lesson must be drawn from the tragedy of Chinese River Dolphin (Lipotes vexillifer), which has already been declared likely extinct. Strong, effective and appropriate protective measures must be carried out quickly to prevent the Yangtze finless porpoise from becoming a second Chinese River Dolphin, and save the biodiversity of the Yangtze River as a whole.

Aquatic resource conservation - The first Yangtze finless porpoise successfully born in captivity

Background. As the sole freshwater subspecies of finless porpoise (Neophocaena phocaenoides), the Yangtze finless porpoise (N. p. asiaeorientalis) lives only in the middle and lower reaches of the Yangtze River and its appended Poyang and Dongting Lakes. As a result of human activity on the river, including over and illegal fishing, pollution, transportation and dam construction, the population of Yangtze finless porpoises has been steadily and rapidly decreasing during the past several decades, which leads the animal to be endangered. Methods. For saving this unique animal from extinction, three corresponding measures, in situ conservation, ex situ conservation, and intensifying breeding research in captivity, were proposed and have been implemented since the 1980s. Results. After successfully rearing the animals in captivity for almost nine years, the first Yangtze finless porpoise was successfully born in captivity on July 5, 2005. The calf is male, with a body length of 69 cm. This is the first freshwater cetacean ever born in captivity. Conclusion. The successful birth of this calf confirms that it is possible to breed the Yangtze finless porpoise in captivity. Furthermore, this will greatly benefit the conservation efforts, and also greatly bolster our on-going efforts to study the reproductive biology of these animals. Recommendation. More studies and efforts are expected to establish a sustainable, captive colony of the Yangtze finless porpoise, which will not only greatly expand our knowledge about the reproduction biology of this animal, but also help to redeem the wild population through a careful yearly 'soft releasing' process.

Range-wide Yangtze freshwater dolphin expedition: The last chance to see Baiji?

Background, Aim and Scope. There are two species of fresh water cetaceans surviving in the Yangtze River system in China: Baiji (Lipotes vexillifer) and Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis). As a result of the expansion of human activities on the river, their distribution ranges appear to be decreasing and in the case of the Baiji, are even being restricted to several sections. The Baiji is the world's most critically endangered cetacean species with a population estimated at only a few tens of individuals. The Yangtze finless porpoise is the world's only freshwater-adapted population of the species, and it has been estimated that only around 1,000 individuals remain in the river system. In order to prevent the extinction of Baiji and a sharp decline in the abundance of the porpoise, in situ conservation (i.e. in the river) and two ex situ conservation (i.e. in semi-natural reserves and in captivity) strategies were proposed and have been implemented since the early 1990s. In view of both the severely endangered status of the animals and the severely degraded conditions of their habitats, the feasibility and actual status of these two strategies are raised for discussion. Main Features. The threats faced by the cetaceans are mainly from the unfettered exploitation of the river's resources. In the past 20 years, five nature reserves have been established along the river. Imposing maximum prohibition of harmful and illegal fishing methods in the reserves might prolong the process of extinction of these cetaceans in the wild, but so far, the administrative measures taken in the reserves have not yet kept the abundance from sharply declining. As human use of the river and its resources is expected to intensify for many decades into the future, the ability of the river to continue to support these species is certainly undecided. Therefore, rescuing animals from the river and establishing viable breeding populations in semi-natural reserves, in which the environment is similar to the main stream of the river, and in captivity, has to be considered urgently as the short-term goal of ex situ strategies. Since the abundance of porpoises is higher than that of the Baiji, we have first established breeding populations of them in the semi-natural reserves and in captivity. But, considering the extremely low density of Baiji in the river, an immediate range-wide Yangtze Baiji survey is an urgent need for locating and capturing sufficient Baiji for successfully establishing a breeding population of them in semi-natural reserves. Results. Two semi-natural reserves (in Shishou, Hubei Province, and Tongling, Anhui Province) have been set up along the river in order to establish breeding populations of the Baiji and the porpoises. So far, several small groups of porpoises that were caught in the main stream of the river have successively been introduced into the semi-natural reserves. Under careful management, these animals in both of the semi-natural reserves not only survive, but can also reproduce naturally and successfully. At least one or three calves were born in each reserve each year. Additionally, a breeding group of porpoises is being established at the Baiji Dolphinarium at the Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan. There are presently four adults and one calf living in the Dolphinarium. The calf, born in July, 2005, is the first captive bred Yangtze Finless Porpoise in the world. In preparation for the range-wide Yangtze cetacean survey, a 9-day pilot expedition on the river near Wuhan was conducted in March, 2006, in order to develop methods for locating the Baiji. No Baiji were expected to be seen in such a short period but about 40 porpoise sightings were observed. Results of the pilot survey indicated that traditional visual and acoustical survey methods for cetaceans should be adapted to find the elusive Baiji in the river. Currently, the range-wide Yangtze cetacean survey is in preparation. The survey will cover over 1,700 km of the Yangtze River from Yichang to Shanghai, and is expected to provide detailed information on Baiji and porpoise numbers and distribution patterns in the river. Discussion. Although the short-term goal of ex situ conservation is to rescue cetaceans from the river and to establish viable breeding populations in semi-natural reserves and in captivity, the long-term goal of releasing the animals back into the river when the threats have decreased and the natural environment has been improved, should not be neglected. Moreover, the in situ conservation efforts in the natural reserves, and even in the entire Yangtze River system, including the lakes, should not be ignored or abandoned at any time. The activities contributing to the conservation of the Baiji and the porpoise in the wild have the incidental effect of benefiting the entire Yangtze ecosystem and other rare threatened species. The dynamics of the groups of porpoises in semi-natural reserves should be monitored continually, in order to guide the establishment of breeding groups of Baiji in these semi-natural reserves in the near future. Conclusions. Under the existing severely degraded conditions of the Yangtze system, the sharply fall populations of Baiji and porpoises will not be suspended in the foreseeable future. Therefore, ex situ conservation should be emphasized, and the severely threatened Baiji in the river should be removed and translocated to semi-natural reserves for establishing viable breeding populations. The successful program of capturing, translocating and maintaining finless porpoises in the Shishou semi-natural reserve has demonstrated its adequacy as an ex situ environment for cetaceans. Following the successful pilot survey in the river, the immediate range-wide Yangtze cetacean survey is proposed and is in preparation. The range-wide survey is expected to ensure that any remaining Baiji can be found reliably and captured successfully after the survey. Recommendations and Perspectives. During the range-wide survey, not only the Baiji but also the porpoise as well as their habitats should be investigated based on visual and acoustical methods that adapted to the river and the animals. Meanwhile, the current risk levels to the Baiji and porpoises should be evaluated at each area where Baiji or porpoises can be reliably sighted. Any capture efforts should be targeted on the most threatened areas, or where there is maximum risk of injury or death. The immediate track of the Baiji should be carried out once a Baiji is sighted during the range-wide survey in order to obtain the movement route of the animals, which is crucial information for the successful capture operation. Additionally, the need to establish new semi-natural reserves for the porpoises should be placed on the agenda of local and central governments in the near future.

Genetic variation in the endangered Anisodus tanguticus (Solanaceae), an alpine perennial endemic to the Qinghai-Tibetan Plateau

We used random amplified polymorphic DNA markers (RAPDs) to assess genetic variation between- and within-populations of Anisodus tanguticus (Solanaceae), an endangered perennial endemic to the Qinghai-Tibetan Plateau with important medicinal value. We recorded a total of 92 amplified bands, using 12 RAPD primers, 76 of which (P = 82.61%) were polymorphic, and calculated values of H-t and H-sp of 0.3015 and 0.4459, respectively, suggesting a remarkably high rate of genetic variation at the species level. The average within-population diversity also appeared to be high, with P, H-e and H-pop values of 55.11%, 0.1948 and 0.2918, respectively. Analyses of molecular variance (AMOVA) showed that among- and between-population genetic variation accounted for 67.02% and 32.98% of the total genetic variation, respectively. In addition, Nei's coefficient of differentiation (G(ST)) was found to be high (0.35), confirming the relatively high level of genetic differentiation among the populations. These differentiation coefficients are higher than mean corresponding coefficients for outbreeding species, but lower than reported coefficients for some rare species from this region. The genetic structure of A. tanguticus has probably been shaped by its breeding attributes, biogeographic history and human impact due to collection for medicinal purposes. The observed genetic variations suggest that as many populations as possible should be considered in any planned in situ or ex situ conservation programs for this species.

Study on the thermal stability of InN by in-situ laser reflectance system

The thermal stability of InN in the growth environment in metalorganic chemical vapor deposition was systematically investigated in situ by laser reflectance system and ex situ by morphology characterization, X-ray diffraction and X-ray photoelectron spectroscopy. It was found that InN can withstand isothermal annealing at temperature as high as 600 degrees C in NH3 ambient. While in N-2 atmosphere, it will decompose quickly to form In-droplets at least at the temperature around 500 degrees C, and the activation energy of InN decomposition was estimated to be 2.1 +/- 0.1 eV. Thermal stability of InN when annealing in NH3 ambient during temperature altering would be very sensitive to ramping rate and NH3 flow rate, and InN would sustain annealing process at small ramping rate and sufficient supply of reactive nitrogen radicals. Whereas In-droplets formation was found to be the most frequently encountered phenomenon concerning InN decomposition, annealing window for conditions free of In-droplets was worked out and possible reasons related are discussed. In addition, InN will decompose in a uniform way in the annealing window, and the decomposition rate was found to be in the range of 50 and 100 nm/h. Hall measurement shows that annealing treatment in such window will improve the electrical properties of InN. (c) 2005 Elsevier B.V. All rights reserved.

圈养大熊猫遗传结构研究

大熊猫(Ailuropoda melanoleuca)是我国特有的濒危野生动物之一，迁地保护已经成为大熊猫物种保护的一个重要方面。当前大熊猫圈养种群数量增长很快，但是其“多雄配一雌”的交配（配种方式），以及生产过程中记录遗失等原因，造成圈养种群普遍存在亲子关系不清、谱系混乱等问题。为了加强遗传管理，有必要进行亲子关系鉴定、完善谱系；还需要检测种群的基因多样性水平，并在此基础上提出相应的遗传管理建议。 本研究应用9个具有高度多态性的大熊猫微卫星标记，对来自成都大熊猫繁育研究基地2006和2007年度出生的17只大熊猫幼崽及其全部候选父母共37个样品做了基因型分析；然后应用最大似然法，判断幼崽的父－子关系。同时，还对来自卧龙大熊猫保护研究中心的31只大熊猫个体也做了基因分型。将两个种群的数据进行比较：1）等位基因多样性和杂合度水平；2）通过F统计法，分析两个种群的遗传分化水平；3）通过遗传距离法，对所有个体进行聚类分析。 研究结果表明： 1）在母子关系不清的情况下，9个微卫星标记联合的父亲鉴定排除概率E为0.940090；而在母子关系确实的条件下，E= 0.993933。由于本研究中所有后代的母亲都是清楚的，因此这9个微卫星位点能够有效用于圈养大熊猫的亲子鉴定。似然法分析也表明，本研究所获得的亲子鉴定结果置信度在95％以上。 2）2005年种源交换后，成都大熊猫的等位基因多样性和杂合度水平都略高于卧龙种群（但没有达到显著水平），两个种群间的遗传分化水平也有所降低。但是，与卧龙相比，成都种群面临较大的近交压力。 基于以上研究结果，我们建议：进一步加强种源交换和基因交流，把两个种群当作一个遗传单元(MU)来进行管理。 Giant panda (Ailuropoda melanoleuca) is one of the endangerd wildlife endemic to China, and the ex-situ breeding become more and more important for the conservation of this speices. Although the captive population is expanding rapidly, the uncertainty occurs because the paternities of cubs are not clear due to the breeding pattern of “multiple male to single female,”as well as the records lost, resulting in errors in the studbook. For this reason, the paternity of the cubs and the genetic diversity of the captive giant pandas should be tested carefully to get information for the genetic management in the future. 9 polymorphism microsatellite markers were used to do paternity assignment for the 17 cubs born in 2006 and 2007 from Chengdu Research Base for Giant Panda Breeding (CGB) based on the maximum-likelihood methods. A total of 37 individuals were sampled, including all the candidate dams and sires. These samples were also used for comparing with 31 individuals sampling from Wolong China Research and Conservation Center for the Giant Panda (WCG). The comparing indexes were: 1) Allelic diversity and heterozygosity; 2) Genetic differentiation based on F-statistic; 3) Cluster analysis based on genetic distance. The results show that: 1) If the mother is unkown, the combined exclusion probability using these 9 loci is 0.940090. If the mother is known then the exclusion probability is 0.993933. Since the dam-offspring relationship is known in captive populations, the results could resolve unknown paternities in the study. And the confidence level of the results is 95% based on the likelihood methods. 2) The allelic diversity and the heterozygosity of CGB were higher than WCG (n ot significant), and the genetic differentiation was reduced a little since the genetic exchange between two populations in 2005. However, the population of CGB will be threatening by inbreeding seriously than that of WCG. From above, we suggest to reiforce the genetic exchange and geneflow between CGB and WCG, and these two populations should be regarded as one genetic management unit (MU).

In situ electrochemical scanning tunnelling microscopy investigation of structure for horseradish peroxidase and its electrocatalytic property

Immobilization of protein molecules is a fundamental problem for scanning tunnelling microscopy (STM) measurements with high resolution. In this paper, an electrochemical method has been proved to be an effective way to fix native horseradish peroxidase (HRP) as well as inactivated HRP from electrolyte onto a highly oriented pyrolytic graphite (HOPG) surface. This preparation is suitable for both ex situ and in situ electrochemical STM (ECSTM) measurements. In situ STM has been successfully employed to observe totally different structures of HRP in three typical cases: (1) in situ ECSTM reveals an oval-shaped pattern for a single molecule in neutral buffer solution, which is in good agreement with the dimension determined as 6.2 x 4.3 x 1.2. nm(3) by ex situ STM for native HRP; (2) in situ ECSTM shows that the adsorbed HRP molecules on HOPG in a denatured environment exhibit swelling globes at the beginning and then change into a V-shaped pattern after 30 min; (3) in situ ECSTM reveals a black hole in every ellipsoidal sphere for inactivated HRP in strong alkali solution. The cyclic voltammetry results indicate that the adsorbed native HRP can directly catalyse the reduction of hydrogen peroxide, demonstrating that a direct electron transfer reduction occurred between the enzyme and HOPG electrode, whereas the corresponding cyclic voltammograms for denatured HRP and inactivated HRP adsorbed on HOPG electrodes indicate a lack of ability to catalyse H2O2 reduction, which confirms that the HRP molecules lost their biological activity. Obviously, electrochemical results powerfully support in situ STM observations.

The Heterogeneous Growth of P(3MeTh)--An Ellipsometric Study

Successive thicker P(3MeTh) layers are analysed by ex situ conventional and imaging ellipsometry. Thin films display a smooth surface, are compact and homogeneous while for a growth charge above 20 mC cm(-2) the polymer structure modifies to a still uniform but less dense layer. A two-layer model is used and a mathematical procedure is developed to obtain, simultaneously, from the experimental ellipsometric parameters, Delta and Psi, the thickness and the complex refractive index of P(3MeTh) films grown up to 80 mC cm(-2). Thicker polymer layers are disordered and present a high degree of surface roughness. 

Prevalence Of Shear Banding In Compression Of Zr41Ti14Cu12.5Ni10Be22.5 Pillars As Small As 150 Nm In Diameter

Recently, the size dependence of mechanical behaviors, particularly the yield strength and plastic deformation mode, of bulk metallic glasses (BMG) has created a great deal of interest. Contradicting conclusions have been drawn by different research groups, based on various experiments on different BMG systems. Based on in situ compression transmission electron microscopy (TEM) experiments on Zr41Ti14Cu12.5Ni10Be22.5 (Vit 1) nanopillars, this paper provides strong evidence that shear banding still prevails at specimen length scales as small as 150 nm in diameter. This is supported by in situ and ex situ images of shear bands, and by the carefully recorded displacement bursts under load control its well as load drops under displacement control. Finite element modeling of the stress state within the pillar shows that the unavoidable geometry constraints accompanying such experiments impart a strong effect on the experimental results, including non-uniform stress distributions and high level hydrostatic pressures. The seemingly improved compressive ductility is believed to be due to such geometry constraints. Observations underscore the notion that the mechanical behavior of metallic glasses, including strength and plastic deformation mode, is size independent at least in Vit 1. (C) 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Study of thin-film SO2 gas sensors by photometric and ellipsometric methods

Pt-, Pd-, and Zr-doped SnO2 thin films and dopant-free VOx films were fabricated by planar magnetron sputtering. Tests for sensitivity to SO2 for all samples were conducted at 180 degreesC, and the sensitivities were investigated ex situ with photometric and ellipsometric methods at room temperature. It was found that the optical sensitivities as well as the sensitive wavelength region for SnO2 films could be tuned by doping. The Pd-doped SnO2 films had good sensitivity in the visible range, and the Zr-doped in the near IR. The dominant sensitive wavelength region for VOx films fell into the visible range, and the ratio of the sensitivity in the visible to that in the near IR increased with O-2/Ar in the depositing atmosphere. (C) 2001 society of Photo-Optical instrumentation Engineers .

Low-temperature growth of InN by MOCVD and its characterization

Metalorganic chemical vapor deposition growth of InN on sapphire substrate has been investigated between 400 degrees C and 500 degrees C to seek the growth condition of InN buffer layer, i.e. the first step of realization of the two-step growth method. Ex situ characterization of the epilayers by means of atomic force microscope, scanning electron microscope and X-ray diffraction, coupled with in situ reflectance curves, has revealed different growth circumstances at these temperatures, and conclusion has been reached that the most suitable temperature for buffer growth is around 450 degrees C. In addition, the growth rate of InN at the optimized temperature with regard to different precursor flow rates is studied at length. (c) 2004 Elsevier B.V. All rights reserved.