颈槽蛇的地理变异

地理变异是蛇类中较为常见的现象。颈槽蛇Rhabdophis nuchalis在中国具有广泛的地理分布,是研究地理变异的理想物种。统计分析采自中国各地的61号颈槽蛇标本外部形态特征,对其地理变异模式进行了研究。结果表明:来自云南省高黎贡山的种群与其他种群在眶后鳞数、腹鳞数、上唇鳞黑纹的数量和后颔片间是否有小鳞等形态特征上的差异有统计学意义。     

Snake venomics and antivenomics of Protobothrops mucrosquamatus and Viridovipera stejnegeri from Taiwan: Keys to understand the variable immune response in horses

The proteomes of the venoms of the snakes Viridovipera stejnegeri and Protobothrops mucrosquamatus from Taiwan were characterized by N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of in-gel generated tryptic peptides. Proteins belonging to the following toxin classes were identified: metalloproteinase, phospholipase A(2) (PLA(2)), serine proteinase, C-type lectin-like, CRISP, l-amino acid oxidase, disintegrin, and peptides (vasoactive and inhibitors of SVMPs). Nine horses were immunized with a mixture of these venoms. All horses developed a satisfactory immune response against lethality of the venom of V. stejnegeri, whereas only three horses reached the accepted neutralizing potency against the venom of P. mucrosquamatus. Antivenoms were prepared from pools of 'good responder' (GR) and 'poor responder' (PR) horses and compared by antivenomics and neutralization tests. A similar neutralizing response was observed between the GR and PR antivenoms against the venom of V. stejnegeri, whereas antivenom from PR had a lower neutralizing activity against effects of P. mucrosquamatus venom than antivenom from GR. The low potency of the plasma of some horses against this venom is a consequence of the low immunogenicity of the neurotoxic PLA(2) trimucrotoxin. Our results provide clues for innovating the immunization scheme to generate improved antivenoms.     

湖南烙铁头蛇毒蛋白组分的双向电泳分析

烙铁头蛇是世界上剧毒的蛇种之一,其所携带的毒素能够导致严重的机体损伤。应用蛋白质双向电泳技术,对湖南烙铁头蛇蛇毒蛋白的蛋白质组分进行分析。通过等电聚焦和SDS-PAGE凝胶电泳分析获得完整的烙铁头蛇毒全蛋白质的图谱,经胶体考马斯亮蓝染色后,应用PDQuest软件对蛋白表达谱进行分析。通过等电聚焦和SDS-PAGE凝胶电泳有83个蛋白质组分被检测出来。其中大约90.00%的蛋白质的相对分子质量(Mr)分布在15～45 kDa之间,大约72.29%的蛋白质等电点(pI)在4.0～7.0之间。通过对烙铁头蛇毒的蛋白组学研究,获得其蛇毒蛋白质组分的表征特点,为后续进一步研究各组分的身份和潜在功能奠定基础,既可以提出新的治疗方案又可以为新的药理应用提供宝贵资源。     

原矛头蝮蛇毒毒性及恢复规律

目的:对原矛头蝮(P.mucrosquamatus)的毒性及其恢复规律进行考察,增进对该蛇毒性的认识。方法:用该蛇连续攻击小鼠,用梯度剂量蛇毒对小鼠进行腹腔注射,统计死亡率,以折算绝对毒性和半数致死量。对毒性耗竭个体,在梯度时间间隔采集蛇毒,以考察毒性恢复规律。结果:原矛头蝮在2 h内可导致6只小鼠死亡。蛇毒对小鼠的半数致死量为12.204μL/Kg。毒性耗竭后,在间隔21天所获得蛇毒的容积达到最大值即190μL,湿重为149.7 mg。结论:毒蛇在攻击猎物时,并不一次性排完毒腺中的所有毒液,随着攻击次数的增多,毒腺中毒液量逐步减少直至耗竭。其毒液量的恢复需要约3周时间,期间,单次排毒量逐渐增加,干物质的含量也基本呈递增趋势。就单次最大排毒量而言,原矛头蝮高于竹叶青、蝮蛇和银环蛇。其单位剂量毒性高于五步蛇和竹叶青。     

莽山烙铁头蛇毒液蛋白质组学研究(英文)

目的收集一条中国特有品种莽山烙铁头蛇的新鲜毒液,对其蛇毒蛋白作生化分析。方法先以电泳胶片法分析比较莽山烙铁头蛇毒与中国大陆及台湾、日本琉球各地其它品种的烙铁头蛇和各地其他品种的烙铁头蛇的粗蛇毒,在凝胶电泳中展现之异同。然后利用高效液相色谱法(HPLC)将莽山烙铁头蛇毒中的磷脂酶分离,再以质谱仪等鉴定,并与以往相同方法分析莽山烙铁头蛇所得到的磷脂酶分析结果比较。结果我们的研究证实了莽山烙铁头蛇毒含唯一主要的磷脂酶,其含量大约占蛇毒总重量的58%,与中国其他烙铁头蛇品种及日本琉球烙铁头蛇的蛇毒某些碱性磷脂酶之氨基酸序列及蛋白结构特别相似。此碱性磷脂酶主要毒性是水肿、局部炎症及肌肉坏死。结论莽山烙铁头蛇咬伤的临床表现为抗凝与出血,局部炎症坏死等特点,亦可由其蛇毒成分得到印证与解释。     

Studies on the Pollen Morphology and Seed Coat of the Genus Cistanche (Orobanchaceae) in China

In the present paper pollen grains of 5 species and seed coat of 4 species of the genus Cistanche (Orobanchaceae) from China were examined by scanning electron microscope As a result, two types of the pollen exine sculpture are distinguished: (1) tuberculate, C. salsa and C. lanzhouensis; (2) rugulate or fine reticulation formed by the fusion of rugae, C. sinensis, C. deserticola and C. tubulosa. Chinese Cistanche was classified into two sections based on the gross morphology by the present author (Zhang, 1984). C. lanzhouensis and C. sinensis were included in one section. This classification is in conflict with the pollen type, which indicates that the characters of pollen grains and external morphology in this genus have evolved at different rates. The differences in pollen morphology of Chinese Cistanche can serve as characters for delimitating species. According to our abservation, C. lanzhouenis Z. Y. Zhang differs from the other members of Cistanche not only in external morphology but also in pollen morphology. Pollen grains are subprolate in this species, while prolate in the other four species, and exine sculpture is tuberculate. The present study provides the establishment of this new species (Zhang, 1984) with palynological evidence. The seed coat sculpture in Chinese Cistanche are constantly alveolate, but there are some slight differences, by which the sculpture can be divided into two types: (1) testa cells have or have not thickenings on the inner anticlinal walls; cavernulous sculpture is present on outer periclinal wall of some cells: C. sinensis; (2) testa cells have striate thickenings on the inner anticlinal walls: C. salsa, C. deserticola and C. tubulosa. The characters of seed coatof some significance for the delimitation of species.     

DID EGG‐LAYING BOAS BREAK DOLLO'S LAW? PHYLOGENETIC EVIDENCE FOR REVERSAL TO OVIPARITY IN SAND BOAS (ERYX: BOIDAE)

Re-evolution of lost complex morphological characters has been proposed for several characters, including insect wings, limbs, eyes in snakes, and digits in lizards, among others. There has also been much interest in whether the transition from oviparity to viviparity is reversible, particularly in squamate reptiles where the transition to viviparity has occurred more times than in any other lineage. Here, we present a phylogenetic analysis of boid snakes based on a concatenated multigene study of all genera of erycines, New and Old World boines, plus other groups thought to be closely related with boines such as monotypic species Calabaria and Casarea . We reconstruct ancestral parity mode on this phylogeny and present statistical evidence that oviparity reevolved in a species of Old World sand boa in the genus Eryx nearly 60 million years after the initial boid transition to viviparity. Remarkably, like other viviparous boas hatchlings of oviparous Eryx lack an egg-tooth providing independent evidence that oviparity is a derived state in these species.     

Isolation and characterization of a novel proteinase inhibitor from the snake serum of Taiwan habu (Trimeresurus mucrosquamatus).

A proteinase inhibitor (designated as TMI) was isolated and purified from the snake serum of Taiwan habu (Trimeresurus mucrosquamatus) by using successive chromatographies which included Sephadex G-100, DEAE-Sephacel chromatographies, and C(4) reverse-phase HPLC. The purified inhibitor was shown to be a homogeneous protein with a molecular mass of about 47 or 36 kDa in the presence or absence of a reducing agent, beta-mercaptoethanol. The inhibitor decreases in molecular mass by about 23% with N-linked neuraminidase treatment, suggesting that it is a glycoprotein. Further enzymatic analyses indicated that this inhibitor possesses strong inhibitory activities toward three zinc-dependent metalloproteinases and not fibrinogenolytic serine proteases previously isolated from the venom of the same snake species with an IC(50) of about 0.2-1.1 microM. Its IC(50) value was approximately three orders of magnitude more effective than those of the tripeptide inhibitors we previously purified from the crude venom of the same snake (Biochem. Biophys. Res. Commun. 248, 562-568 (1998)). The purified inhibitor showed stronger inhibitory action against caseinolytic activities of crude venoms from closely related species of Taiwan habu than those from unrelated species. N-terminal sequence analysis showed that its sequence is distinctly different from sequences of those serum inhibitors reported for other snake species in the literature. Based on inhibition susceptibility and primary structures of various snake protease inhibitors, it is suggested that this novel inhibitor isolated from the serum of Taiwan habu may be a unique self-defense protein factor mainly for protection against envenomation from snakes of the same genus.     

Completion of the life cycle of Sarcocystis zuoi , a parasite from the Norway rat, Rattus norvegicus

Transmission experiments were performed to elucidate the life cycle of Sarcocystis zuoi found in Norway rats ( Rattus norvegicus ) in China. Two king rat snakes ( Elaphe carinata ) fed sarcocysts from the muscles of 4 naturally infected Norway rats shed sporocysts measuring 10.8 ± 0.7 × 8.0 ± 0.7 μm, with a prepatent period of 8-9 days. Sporocysts from the intestine of 2 experimentally infected king rat snakes were given to the laboratory Sprague-Dawley (SD) rats ( R. norvegicus ) and Kunming (KM) mice ( Mus musculus ). Microscopic sarcocysts developed in the skeletal muscles of SD rats. No sarcocysts were observed in KM mice. Characters of ultrastructure and molecule of sarcocysts from SD rats were confirmed as S. zuoi . Our results indicate that king rat snake is the definitive host of S. zuoi .     

亚洲蝮亚科蛇属间系统发生支序分析 (蛇亚目：蝰科)

在大量比较形态研究的基础上,选择了５个特征方面２８个性状,以支序分析方法探讨了分布于亚洲蝮亚科蛇５属１４种的系统发生关系,结果表明：该亚科蛇类可以划分为３个不同的类群,第一个类群包括尖吻蝮属、瘤鼻蝮属、红口腹属,它们具有较多的祖征,代表了该科中原始一类,基中红口蝮可能是最原始的一属;亚洲蝮属单独形成一个类群;第三个类群为原广义的烙铁头蛇属,包括竹叶青蛇属、原矛蝮属、烙铁头蛇属、黑绿烙铁头蛇属、莽山     

Integrative taxonomy provides evidence for the species status of the Ibero‐Maghrebian grass snake Natrix astreptophora

The grass snake (Natrix natrix) is Europe's most widely distributed and, in many regions, most common snake species, with many morphologically defined subspecies. Yet, the taxonomy of grass snakes is relatively little studied and recent work has shown major conflicts between morphologically defined subspecies and phylogeographical differentiation. Using external morphology, osteological characters, and information from 13 microsatellite loci and two mitochondrial markers, we examine differentiation of the subspecies N. n. astreptophora from the North African Maghreb region, the Iberian Peninsula and neighbouring France. According to previous studies, N. n. astreptophora corresponds to a deeply divergent mitochondrial clade and constitutes the sister taxon of all remaining grass snakes. In the French Pyrenees region, there is a contact zone of N. n. astreptophora with another subspecies, N. n. helvetica. Our analyses of microsatellites and mitochondrial DNA reveal that the distribution ranges of the two taxa abut there, but both hybridize only exceptionally. Even though many morphological characters are highly variable and homoplastic in grass snakes, N. n. astreptophora differs consistently from all other grass snakes by its reddish iris coloration and in having significantly fewer ventral scales and another skull morphology. Considering further the virtual absence of gene flow between N. n. astreptophora and N. n. helvetica, and acknowledging the morphological distinctiveness of N. n. astreptophora and its sister group relationship to all remaining subspecies of grass snakes, we conclude that Natrix astreptophora (Seoane, 1884) should be recognized as a distinct species. Further research is needed to explore whether N. astreptophora is polytypic because a single sample of N. astreptophora from Tunisia turned out to be genetically highly distinct from its European conspecifics.     

野生烟草种子活力检测及物候特性观察

在烟草属（Nicotiana L．）中,野生种具有抗病、抗虫及抗逆性强等性状,是烟草抗病、抗逆育种的珍贵材料。研究野生烟草种子活力及其物候特点,对于解决烟草远缘杂交过程中的花期相遇问题以及开展抗性鉴定等工作具有重要意义,是烟草抗性育种工作的基础。     

Phylogenetic relationships of North American garter snakes (Thamnophis) based on four mitochondrial genes: how much DNA sequence is enough?

The clade of garter snakes (Thamnophis) includes some of the most abundant and well-studied snakes in North America. However, phylogenetic relationships within this group have been little studied. We used DNA sequences of four mitochondrial genes (cytochrome b and NADH dehydrogenase subunits 1, 2, and 4) to estimate relationships among 29 of the 31 recognized species of Thamnophis plus the related species Adelophis foxi. Both maximum parsimony (MP) and maximum-likelihood (ML) analyses of all these genes combined produced well-resolved trees with moderate (70-89%) to strong (90-100%) bootstrap support for most clades. MP and ML trees were very similar, with no strongly supported conflict between the two analyses. These analyses identify a clade of 12 species largely restricted to México (the "Mexican clade"), and a clade containing 15 species that collectively range from Central America to southern Canada (the "widespread clade"). These two groups are identified as sister taxa in both MP and ML analyses. A clade consisting of the ribbon snakes (T. sauritus and T. proximus) and the common garter snake (T. sirtalis) is placed as the sister group to all other Thamnophis (i.e., the Mexican + widespread clades) in our analyses. High bootstrap proportions at several levels in the tree support the inclusion of both Thamnophis validus, which has traditionally been placed in the genus Nerodia, and the poorly known species Adelophis foxi within Thamnophis. We used randomly sampled characters (i.e., standard bootstrapping) and randomly sampled contiguous blocks of characters to examine the effect of number of characters on resolution of and support for relationships within Thamnophis using MP. In general, these analyses indicate that we have reached a point of strongly diminishing returns with respect to the effect of adding mtDNA sequence characters for the current set of taxa; our sample of 3809 mtDNA characters is apparently "enough." The next steps to improve the phylogenetic estimate may be to add nuclear DNA sequences, morphology, or behavior, or to sequence additional mtDNA lineages within species.     

A new species of Serpentirhabdias Tkach,Kuzmin & Snyder, 2014 (Nematoda: Rhabdiasidae) parasitic in the brown ground snake Atractus major Boulenger (Reptilia: Serpentes: Dipsadidae) in Brazil

Serpentirhabdias atracti n. sp. is described based on specimens discovered in the lung of Atractus major Boulenger from Caxiuanã National Forest, Pará, Brazil. The new species is assigned to Serpentirhabdias Tkach, Kuzmin & Snyder, 2014 based on morphological characters (comparatively thin body cuticle without prominent inflations, arrangement of circumoral papillae in two lateral groups, pre-equatorial position of vulva, eggs in uteri at early cleavage stages), as well as because of its parasitism in snakes. The new species is most similar to S. vellardi (Pereira, 1928) due to the absence of lips and buccal capsule, similar body dimensions, and the specificity to dipsadid snakes in Brazil. The two species differ in the shape of the tail (bulbous dilatation in the posterior part followed by a thread-like tail tip present in S. atracti n. sp.), the width of the oesophagus, and the size of the excretory glands. Serpentirhabdias atracti n. sp. is the sixth species of this genus found in the Neotropical Region.     

An exotic species is the favorite prey of a native enemy

Although native enemies in an exotic species'' new range are considered to affect its ability to invade, few studies have evaluated predation pressures from native enemies on exotic species in their new range. The exotic prey naiveté hypothesis (EPNH) states that exotic species may be at a disadvantage because of its naïveté towards native enemies and, therefore, may suffer higher predation pressures from the enemy than native prey species. Corollaries of this hypothesis include the native enemy preferring exotic species over native species and the diet of the enemy being influenced by the abundance of the exotic species. We comprehensively tested this hypothesis using introduced North American bullfrogs (Lithobates catesbeianus, referred to as bullfrog), a native red-banded snake (Dinodon rufozonatum, the enemy) and four native anuran species in permanent still water bodies as a model system in Daishan, China. We investigated reciprocal recognition between snakes and anuran species (bullfrogs and three common native species) and the diet preference of the snakes for bullfrogs and the three species in laboratory experiments, and the diet preference and bullfrog density in the wild. Bullfrogs are naive to the snakes, but the native anurans are not. However, the snakes can identify bullfrogs as prey, and in fact, prefer bullfrogs over the native anurans in manipulative experiments with and without a control for body size and in the wild, indicating that bullfrogs are subjected to higher predation pressures from the snakes than the native species. The proportion of bullfrogs in the snakes'' diet is positively correlated with the abundance of bullfrogs in the wild. Our results provide strong evidence for the EPNH. The results highlight the biological resistance of native enemies to naïve exotic species.     

副沙鳅属系统发育分析

以沙鳅属Botia为外类群,共发现了副沙鳅属Parabotia 35个有系统发育意义的外部与骨骼形态特征,并由此重建副沙鳅属系统发育关系为:漓江副沙鳅 (双斑副沙鳅 (花斑副沙鳅 (武昌副沙鳅 点面副沙鳅)))或漓江副沙鳅 (双斑副沙鳅 (武昌副沙鳅 (点面副沙鳅 花斑副沙鳅))).具有囟门;后翼骨外侧面为不规则的梯形;吻骨侧面观呈一长方形;无下舌软骨;具有3对咽鳃骨;中喙骨弯曲较大,近直角;上吻皮中间有切刻状缺口,左右不连续;以及颐部无纽状突起等构成了副沙鳅属的共同离征.     

Presence of the large aquatic snake Palaeophis africanus in the middle Eocene marine margin of the Congo Basin,Cabinda, Angola

Ten isolated snake vertebrae from Landana and Sassa-Zao, Cabinda Exclave, Angola, present a “primitive” grade morphology with a weak lateral compression and do not belong to Palaeophis aff. typhaeus as originally referred to. They well belong to a single taxon and are here attributed to Palaeophis africanus for which the intracolumnar variation is described and illustrated. This species is Lutetian (middle Eocene) in age and originates from a marine coastal environment confirming again the aquatic capabilities of palaeophiid snakes. It represents the third largest species of Palaeophis with P. colossaeus and P. maghrebianus to which it is closely related in our tentative phylogenetic analysis, indicating that these three taxa could belong to an African clade. This study also contributes to the debate on the existence of primitive and advanced grades among palaeophiid snakes. Palaeophis presents laterally compressed anterior trunk vertebrae that could have been often erroneously considered as representing advanced grade species and potential parataxonomy.     

Morphological and mitochondrial DNA characters for identification and phylogenetic analysis of the myiasis-causing flesh fly Wohlfahrtia magnifica and its relatives, with a description of Wohlfahrtia monegrosensis sp. n. Wyatt & Hall

Abstract.  Wohlfahrtia magnifica (Schiner) (Diptera: Sarcophagidae) is a major cause of traumatic myiasis in livestock in Central and Eastern Europe and in countries bordering the Mediterranean. The present study explored the utility of external body characters, genitalia characters and mitochondrial DNA characters for identification of this and related species in the subfamily Paramacronychiinae. Sequence analyses of the 3' terminal 273 bp of the mitochondrial cytochrome b gene revealed two lineages of W. magnifica , one from Spain and France and the other from the rest of Eurasia, differing by only two base pairs. Phylogenetic analysis of cytochrome b showed that W. magnifica and Wohlfahrtia vigil Walker were sister species; this conclusion was not contradicted by a phylogenetic analysis of the morphological characters. Based on cytochrome b , the genetic distance between specimens of W. vigil from Europe and North America was sufficiently large to justify the recognition of more than one species. A new species, Wohlfahrtia monegrosensis , from northern Spain, was described, based on morphology and cytochrome b . A unique combination of external body characters of males or females were diagnostic for W. magnifica , the W. vigil group and Wohlfahrtia bella , but only the genitalia characters were diagnostic for all nine species studied.     

中国豹蛛属枯藤豹蛛种组(蜘蛛目,狼蛛科)分类研究

记述了我国3种豹蛛,根据其外雌器和触肢器结构特点,应归入枯藤豹蛛Pardosa xerampelina种组.其中新种1种为蝶形豹蛛Parodsa papilionacasp.nov;另两种为我国新纪录种:草豹蛛P.herbosa Jo et Paik,1984及淡豹蛛P.indecora L.Koch,1879.编制了该种组我国分布种的检索表.模式标本及其他检视标本均保存在中国科学院动物研究所.