Identification of reproductively isolated lineages of Amur grayling (Thymallus grubii Dybowski 1869): concordance between phenotypic and genetic variation

We analysed variation at maternally (mitochondrial DNA control region sequences) and bi-parentally (10 microsatellites) inherited genetic markers, as well as across 12 meristic characters in 7 populations of Amur grayling, Thymallus grubii, from eastern Siberia. All three data sets were concordant in supporting the existence of three diagnosable, reciprocally monophyletic, and most probably reproductively isolated, lineages of grayling within the Amur drainage. There was a significant correlation between genetic and phenotypic divergence, both within and among lineages. Two phenotypically distinct forms (with and without an orange spot on the posterior portion of the dorsal fin), found in sympatry in the lower Amur, most likely result from secondary contact, as they demonstrate 4.6% sequence divergence at the mitochondrial DNA control region. This divergence, together with the existence of at least one nearby population of orange spot grayling outside the Amur drainage (0.8% divergence) underscore the palaeo-hydrological complexity of the system, which presumably promoted genetic divergence in a shifting allopatric framework throughout the Pleistocene. Grayling from the upper Amur, corresponding to the type locality for the species, formed a sister group (1.4-1.6% divergent) to the orange spot lineage perhaps diverging in the early Pleistocene (1.4-1.6 Ma).     

Killed Saccharomyces cerevisiae Protects Against Lethal Challenge of Cryptococcus grubii

Heat-killed Saccharomyces cerevisiae (HKY) vaccination protects mice against aspergillosis, coccidioidomycosis, mucormycosis, or candidiasis. We studied HKY protection against murine cryptococcosis. Once weekly subcutaneous HKY doses (S, 6 × 10⁷; 2S, 1.2 × 10⁸; 3S, 2.4 × 10⁸) began 28 (×3), 35 (×4), or 42 (×6) days prior to intravenous Cryptococcus grubii infection. Survival through 28 days, and CFU in the organs of survivors, were compared to saline-vaccinated controls. In the initial experiment, S, S×4, or 2S reduced brain CFU; liver or spleen CFU was reduced by S×4 or 2S. In a more lethal second experiment, 2S×6, 2S, or 3S×4 improved survival, and HKY regimens reduced CFU in the brain, liver, or spleen, with 2S×6, 2S, or 3S×4 most efficacious. Dose size appears more important than the number of doses: Regimens >S were superior, and 2S and 2S×6 were equivalent. 2S and 3S were equivalent, suggesting doses >2S do not provide additional protection. HKY protects against Cryptococcus, supporting components of HKY as a basis for the development of a panfungal vaccine.     

半圈养条件下东北虎繁殖期的行为时间分配

1998年 10月～ 2 0 0 0年 10月在黑龙江东北虎林园 ,采用随机取样法和瞬时扫描取样法对繁殖期东北虎的行为进行了研究。结果表明 ,在繁殖期的不同阶段东北虎的行为发生较大的变化。在发情交配期雌雄虎的时间分配相似 (P >0 0 5 ) ,均将最多的时间用于运动 ,用于休息的时间较少 ,用于发情交配和其它行为的时间较多。雌虎在妊娠期较多的时间用于休息和运动 ,取食、站立、社会和其它行为的时间均较少 ;产仔期最多的时间用于休息 ,其次是运动 ,用于产仔和其它行为的时间较多 ,用于取食的时间最少 ;哺乳期最多的时间用于休息 ,其次是哺乳和运动 ,用于取食和其它行为的时间较少。活动规律方面 ,各种行为在强度、出现和持续的时间上存在较大差异。单因素方差分析表明 ,在繁殖期的不同阶段雌性东北虎的休息和运动存在极显著差异 (P <0 0 0 1) ,取食存在显著差异 (P <0 0 5 ) ,只有其它行为不存在显著差异(P >0 0 5 )。 10 0日龄内幼虎用于休息的时间最多 ,其次是运动和取食 ,其它行为的时间最少。     

黑龙江黑斑狗鱼线粒体控制区的遗传变异

测定了黑龙江抚远江段和虎林江段黑斑狗鱼(Esoxreicherti)线粒体控制区5'端的560bp序列,在12尾黑斑狗鱼序列中未检测到任何变异位点,即所有个体的序列均完全一致,共享同一种单倍型,群体内和群体间的核苷酸多样性均为0.黑龙江黑斑狗鱼线粒体DNA如此贫乏的遗传多样性,一方面可能是黑龙江水系黑斑狗鱼在更新世冰期可能曾遭受过严重的瓶颈效应;另一方面则可能因目前是国内黑斑狗鱼种群数量锐减,种质资源急剧衰退,导致种群遗传多样性的丧失.建议广泛取样调查不同地理种群,并采用AFLP和微卫星等分子标记检测遗传多样性,以确定遗传变异丰富的黑斑狗鱼种群加以保护,促进黑龙江流域黑斑狗鱼资源的有效保护和可持续利用.     

Three galactose inducible promoters for use in C. neoformans var. grubii

Cryptococcus neoformans is the causative agent of cryptococcal meningoencephalitis, most frequently occurring in immunocompromised individuals. There are three varieties of C. neoformans, var. grubii, var. neoformans, and var. gatti. Worldwide var. grubii is the most prevalent clinical isolate. However, few tools for the study of essential genes in var. grubii exist. Here we describe three endogenous inducible promoters for use in the study of this important opportunistic pathogen. We identified eight potential homologs of S. cerevisiae galactose genes in var. grubii. We found that GAL1, GAL7, and UGE2 were regulated by glucose and galactose and can be used successfully during mating. Our analysis indicated these promoters should prove to be excellent tools for analysis of genes in var. grubii.     

Endemic sturgeons of the Amur River: kaluga, Huso dauricus, and Amur sturgeon, Acipenser schrenckii

General biological characteristics and the contemporary status of the kaluga, Huso dauricus, and Amur sturgeon, Acipenser schrenckii, are described. Both inhabit the Amur River basin. Kaluga is the largest freshwater fish in this river system reaching more than 5.6 m in length and more than 1000 kg in weight. We recognize four populations of kaluga: the first is from the estuary of the Amur River and coastal brackish waters of the Sea of Okhotsk and Sea of Japan, the second is from the lower Amur River, the third is from the middle-Amur, and the fourth occurs in lower reaches of the Zeya and Bureya rivers. Freshwater and brackish water morphs exist in the estuary population, with the freshwater morph predominating in number. The number of individuals in the lower Amur River population at age 2 or greater was recently estimated to be 40 000, and in the middle Amur, 30 000. The population will continue to decline because of rampant overfishing. The Amur sturgeon is represented in the Amur River basin by two morphs: brown and gray. Brown morphs occur in the middle and lower parts of the Amur River; they grow more slowly than the gray ones. Today, the lower Amur River population of Amur sturgeon is made up of 95 000 fish at age 2 or greater and is approximately half as large as the population in the middle Amur River. Populations of kaluga and Amur sturgeon in the Zeya and Bureya rivers are extremely small and on the verge of extinction.     

Morphology of monogeneans parasitizing Amur River fish

Morphology of Bivaginagyrus obscurus from Pseudorasbora is described. B. obscurus differs from species of the genus Dacfylogyrus by 2 vaginae, shape of median hooks, seat of gill filament and length of additional ventral bar. Morphology of Silurodiscoides infundibulavagina and some other species of this genus parasitic on Parasilurus asotus is described. The morphology of Pseudoancylodiscoides from Pseudobagrus fulvidraco is characterized by 2 ventral bars and peculiar seminal vesicle. Morphology of Bychowskyella pseudobagri from Pseudobagrus fulvidraco is described. The testis in this type species is located more ventrally than the ovary.     

吉林珲春自然保护区东北虎和东北豹及其有蹄类猎物的多度与分布

珲春国家级自然保护区是东北虎(Panthera tigris altaica)、东北豹(Panthera pardus orientalis)等濒危物种在中国的核心分布区。为了探究该区域野生动物的多度水平和空间分布, 了解人类干扰情况, 我们运用相对多度指数(relative abundance index, RAI)分析了2013年4-6月设置于此的83个红外相机位点的监测数据。红外相机的总捕获天数6,060 d, 共捕获10科18种野生哺乳动物, 其中鼬科4种, 猫科动物3种, 犬科、鹿科和松鼠科各2种, 猪科、熊科、麝科、猬科和兔科各1种。研究期间共拍摄到东北虎11只个体, 东北豹13只个体。从相对多度指数来看, 东北虎的相对多度(0.84)远高于东北豹(0.48), 它们的有蹄类猎物中梅花鹿(Cervus nippon)的相对多度最高(2.18), 其次为狍(Capreolus pygargus)(1.53)和野猪(Sus scrofa)(0.92)。人类活动和放牧的相对多度水平(分别为40.64和2.76)显著高于野生动物。在空间分布上, 东北虎和梅花鹿主要在保护区的核心区分布, 且与保护区社区共管区的多度水平差异显著, 而东北豹在不同功能区之间的分布差异不显著, 狍在保护区北部的多度水平较高, 但各功能区之间差异不显著, 野猪在社区共管区的多度水平显著高于核心区。可见, 核心区频繁的人类活动和放牧活动对野生动物的保护产生了影响, 未来应加强关于人类干扰对虎、豹种群及其有蹄类猎物的影响评估。     

湿润亚热带峰丛洼地岩溶土壤系统中碳分布及其转移

以桂林丫吉村岩溶试验场为例,研究了湿润亚热带峰丛洼地表层岩溶带生物量碳库、凋落物碳库、土壤有机碳库(SOM)及其活泼性、有机碳分解速率、土壤中CO₂浓度和土壤呼吸CO₂排放,表明岩溶系统中丰富的碳库提供了系统中CO₂的来源,并用D¹³C证实春夏岩溶活跃季节中岩溶输出C约60%来自土壤CO₂.由此认为,驱动岩溶作用的CO₂并非直接来自大气CO₂,而是大气-植物-土壤-水碳素转移的结果,因而揭示了土壤作为碳环境地球化学界面对于表生带岩溶作用的驱动意义.     

人工饲养东北虎幼虎的行为时间分配

1 999年 4月～ 2 0 0 0年 4月 ,采用随机取样法和瞬时扫描取样法对黑龙江东北虎林园的 30只幼虎的行为时间分配及活动规律进行了研究。结果表明 ,幼虎用于运动和卧息的时间较多 ,为 ( 32 .0 8± 1 2 .39) %和 ( 2 8.0 2± 1 0 .76) % ;用于睡眠、站立和嬉戏的时间次之 ,分别为 ( 1 1 .9± 2 5 .2 1 ) %、( 9.0 6± 4.71 ) %和( 1 4.49± 8.72 ) % ;其它行为最少 ,为 ( 4 .66± 3.2 8) %。步行区幼虎园和幼虎园的幼虎活动规律较为相似 ,与育成虎园的幼虎相差较大。单因素方差分析表明幼虎不同个体间不存在显著差异 ,不同年龄组间的运动、卧息和嬉戏差异极显著 ,睡眠差异显著 ,站立和其它行为差异不显著。     

Hydroxyurea Enhances Post-Fusion Hyphal Extension During Sexual Development in C. neoformans var. grubii

Mating and sexual development in C. neoformans var. grubii strains of the H99 background is often less robust than that laboratory generated isogenic C. neoformans var. neoformans strains in the JEC21 background. In Candida albicans and Saccharomyces serevisiae, slowing of DNA synthesis and engagement of the replication stress response, such as that caused by treatment with hydroxyurea (HU), induces filamentation and pseudohyphal growth, respectively. In this study, we investigated the effect of HU treatment on C. neoformans var. grubii morphogenesis. Treatment with HU did not induce filamentation of yeast cells either in liquid culture or on solid YPD or V8 agar. In the presence of the opposite mating partner, we observed early emergence of hyphae in the presence of HU. Semi-quantitative analysis of fusion using marked strains demonstrated that no significant enhancement of fusion in the presence of HU. Transfer of fusion colonies from crosses performed in the absence of HU to V8 + HU revealed enhanced hyphal growth in the presence of HU. Analysis of expression of the target of HU, ribonucleotide reductase, revealed that a phylogenetically divergent catalytic subunit is replication stress responsive in C. neoformans. These results suggest that induction of replication stress promotes post-fusion hyphal growth of C. neoformans var. grubii strains in the H99 background.     

Identification of Novel Hybrids Between Cryptococcus neoformans var. grubii VNI and Cryptococcus gattii VGII

Cryptococcus neoformans and Cryptococcus gattii are pathogenic yeasts causing meningoencephalitis in immunocompromised and immunocompetent hosts. The fungus is typically haploid, and sexual reproduction occurs normally between individuals with opposite mating types, α and a. C. neoformans var. grubii (serotype A) is comprised of molecular types VNI, VNII, and VNB, and C. neoformans var. neoformans (serotype D) contains the molecular type VNIV. Additionally, diploid or aneuploid AD hybrids (VNIII) have been reported. C. gattii contains the molecular types VGI, VGII, VGIII, and VGIV, which encompass both serotypes B and C. To identify possible hybrid strains, URA5-RFLP analysis was performed on 350 globally obtained clinical, environmental, and veterinary isolates. Four clinical isolates from cerebrospinal fluid showed combination patterns of C. neoformans var. grubii and C. gattii: Brazil (n = 2), Colombia (n = 1), and India (n = 1). These strains were monokaryotic and diploid or aneuploid. M13 PCR fingerprinting showed that they contained fragments of both proposed parental groups. Luminex IGS genotyping identified these isolates as hybrids with two different molecular type combinations: three VNI/VGII and one VNI/VGI. Blue color development on CGB agar was delayed in three isolates and absent in one. C. gattii-specific PCR confirmed the presence of C. gattii in the hybrids. CAP59 allele-specific PCR revealed that all the hybrids contained both serotype A and B alleles. Determination of mating-type allelic patterns by PCR revealed that the isolates were αA aB. This is the first study discovering novel natural hybrids between C. neoformans molecular type VNI and C. gattii molecular type VGII.     

Evidence that the human pathogenic fungus Cryptococcus neoformans var. grubii may have evolved in Africa

Most of the species of fungi that cause disease in mammals, including Cryptococcus neoformans var. grubii (serotype A), are exogenous and non-contagious. Cryptococcus neoformans var. grubii is associated worldwide with avian and arboreal habitats. This airborne, opportunistic pathogen is profoundly neurotropic and the leading cause of fungal meningitis. Patients with HIV/AIDS have been ravaged by cryptococcosis--an estimated one million new cases occur each year, and mortality approaches 50%. Using phylogenetic and population genetic analyses, we present evidence that C. neoformans var. grubii may have evolved from a diverse population in southern Africa. Our ecological studies support the hypothesis that a few of these strains acquired a new environmental reservoir, the excreta of feral pigeons (Columba livia), and were globally dispersed by the migration of birds and humans. This investigation also discovered a novel arboreal reservoir for highly diverse strains of C. neoformans var. grubii that are restricted to southern Africa, the mopane tree (Colophospermum mopane). This finding may have significant public health implications because these primal strains have optimal potential for evolution and because mopane trees contribute to the local economy as a source of timber, folkloric remedies and the edible mopane worm.     

Distribution,abundance, and size structure of Amur kaluga <Emphasis Type="Italic">Acipenser dauricus</Emphasis> and Amur sturgeon <Emphasis Type="Italic">A. schrenckii</Emphasis> in the Lower Amur and Amur Estuary

Distribution, population structure, and abundance of Amur kaluga Acipenser dauricus and Amur sturgeon A. schrenckii in the Lower Amur and Amur Estuary have been considered based on the materials of two surveys performed in 2011. Both species occur along almost the entire length of the Lower Amur and in major part of the Amur Estuary. The density of their aggregations has significantly decreased in most regions of the river if compared to the 1960s. In the Amur Estuary, A. dauricus and A. schrenckii are most abundant in freshwater and light salted water bodies of its western part. In 2011, the abundance and biomass of A. dauricus and A. schrenckii in the Lower Amur and Amur Estuary was 345000 specimens and 7110 tons and 289000 specimens and 1946 tons, respectively.     

A new subspecies of the amur grayling Thymallus grubii flavomaculatus ssp. nova (Thymallidae)

A new subspecies of the Amur grayling is described, the yellow-spotted grayling Thymallus grubii flavomaculatus ssp. n. populating the upper reaches of large tributaries of the Lower Amur basin and some rivers flowing to the Tatar Strait, the Sea of Okhotsk, and the Sea of Japan. From nominative subspecies T. g. grubii populating the basin in the upper reaches of the Amur and from other representatives of the genus Thymallus, this subspecies differs in the dorsal fin pattern whose principal trait is a yellow spot on the last two-five interray membranes, in the body coloration, and in some morphological characters. The yellow-spotted grayling in the upper reaches of tributaries sympatrically coexists with the Lower Amur grayling Thymallus sp. No specimens with intermediate characters are found. The sympatric forms spawn at different times, thus being isolated reproductively. Isolation of the yellow-spotted grayling as a subspecies of the Amur grayling T. grubii is confirmed by the results of molecular-genetic investigations.     

Silica spicules and axial filaments of the marine sponge Stelletta grubii (Porifera,Demospongiae)

Summary In all cases an organic axial filament within the silica spicules of Stelletta grubii forms the core of the major axes of the glass. In the small, star-shaped silica spicules (asters) the filament is shown for the first time to be radial with an enlarged center; in the large four-rayed spicules (triaenes) it is four-rayed; and in the large single-rayed spicules (oxeas) the filament is single-rayed. In situ, the filament is not dissolved by boiling nitric acid and thus is apparently protected by encasement within the glass which can also be stratified. The small silica asters are formed by single cells which resemble the so-called spherulous cells of other sponges. The very large size of triaenes and oxeas suggests that they may possibly be formed by more than one cell. The diameter of the filament in the much smaller asters is much narrower than the filament in the larger spicules, indicating a possible relationship between filament diameter and spicule diameter. While the axial filament in larger spicules frequently has a triangular cross-section it can also be hexaognal. Some aster filaments also retain a close to hexagonal cross-section. Filaments freed from large spicules by hydrofluoric acid display a complex morphology; possibly there is an internal silicified core. Some reported aspects of filament morphology are, however, probably artefacts of desilicification with hydrofluoric acid. Offprint requests to: T.L. Simpson, Department of Biology, University of Hartford, West Harford, Connecticut 06117, USA (Permanent affiliation)     

Comparative Ontogenetic Behavior and Migration of Kaluga, Huso Dauricus, and Amur Sturgeon, Acipenser Schrenckii, from the Amur River

We conducted laboratory experiments with kaluga, Huso dauricus, and Amur sturgeon, Acipenser schrenckii, to develop a conceptual model of early behavior. We daily observed embryos (first life phase after hatching) and larvae (period initiating exogenous feeding) to day-30 (late larvae) for preference of bright habitat and cover, swimming distance above the bottom, up- and downstream movement, and diel activity. Day-0 embryos of both species strongly preferred bright, open habitat and initiated a strong, downstream migration that lasted 4 days (3 day peak) for kaluga and 3 days (2 day peak) for Amur sturgeon. Kaluga migrants swam far above the bottom (150cm) on only 1 day and moved day and night; Amur sturgeon migrants swam far above the bottom (median 130cm) during 3 days and were more nocturnal than kaluga. Post-migrant embryos of both species moved day and night, but Amur sturgeon used dark, cover habitat and swam closer to the bottom than kaluga. The larva period of both species began on day 7 (cumulative temperature degree-days, 192.0 for kaluga and 171.5 for Amur sturgeon). Larvae of both species preferred open habitat. Kaluga larvae strongly preferred bright habitat, initially swam far above the bottom (median 50–105cm), and migrated downstream at night during days 10–16 (7-day migration). Amur sturgeon larvae strongly avoided illumination, had a mixed response to white substrate, swam 20–30cm above the bottom during most days, and during days 12–34 (most of the larva period) moved downstream mostly at night (23-day migration). The embryo–larva migration style of the two species likely shows convergence of non-related species for a common style in response to environmental selection in the Amur River. The embryo–larva migration style of Amur sturgeon is unique among Acipenser yet studied.