基于动物学、临床医学跨界研究快速准确诊断1例不明蛇类咬伤的分析

目的 探讨动物学、临床医学跨界研究成果联合运用于快速准确诊断不明蛇类咬伤中的作用及效果.方法 通过回顾性分析1例不明蛇类咬伤患者的临床资料,从病史、动物学生活习性、咬痕鉴别、流行病学、临床表现、蛇类动物学分布6个方面进行剖析不明蛇类咬伤的诊断思路与方法.结果 该例蛇伤患者的病史特征以及诊断与鉴别诊断:(1)病史特征符合...     

广东省养蛇业的现状与发展策略的研究

目的探讨广东省蛇类养殖业的现状与发展的策略方法在全省范围内按各市县养殖场逐个进行实地考查,调查蛇类养殖场的分布、规模、面积、养殖技术和投资金额以及蛇的种类、数量、来源和贸易情况,分析蛇类养殖存在的问题和经营利用的可能性。并以表格的形式搜集各养蛇场的基础资料、近年来的养殖情况及意见和建议。结果3个养殖最成功的蛇场饲养的主要蛇种类有眼镜蛇、眼镜王蛇、滑鼠蛇、乌梢蛇,目前已基本上解决了蛇类养殖的难题,掌握了蛇类冬眠和管理等关键技术,蛇类存栏量都达到10万条以上,存栏的个体都在第三代以上;养殖成本明显下降,经济效益可观。结论眼镜蛇、眼镜王蛇、滑鼠蛇、乌梢蛇是人工养殖成功的种类,提出发展我国蛇类产业的具体措施和策略,为蛇类养殖产业化、规模化和集约化提出合理化的建议。     

乌梢蛇血清的抗出血因子:一个有前途的抗蛇毒药物原料

用柱层析和聚丙烯酰胺凝胶盘状电泳法,从乌梢蛇血清中分离纯化了一个抗出血因子。用SDS-聚丙烯酰胺凝胶电泳法测得其分子量大约为65 kD;测定了五种蝮亚科蛇毒(尖吻蝮、竹叶青蛇、原矛头蝮、哈扑和短尾蝮)的最小出血剂量和乌梢蛇血清中抗出血因子对这五种蛇毒的抗出血活性;还测定了七种蛇毒(除上述五种毒蛇外,还包括圆斑蝰和银环蛇)的半数致死量,以及抗出血因子对中毒小鼠的治疗作用。结果显示:从乌梢蛇血清中提纯的抗出血因子的抗蛇毒活性,不仅可以抵抗它的捕食者尖吻蝮的蛇毒,而且还可以抵抗具出血活性的其它蛇毒;但它对不具出血活性的银环蛇毒的致死抑制作用不明显。该抗出血因子不仅在体外实验表现出强的中和出血毒素的活性,而且在体内实验中亦表现出对中毒小鼠良好的治疗作用,因而可能成为新的抗蛇毒药物的有前途的原料。乌梢蛇血清对血循毒的中和能力的获得,可能归因于尖吻蝮与乌梢蛇之间捕食与被捕食相互作用的关系。     

几种游蛇的Cyt b基因片段序列分析及其演化关系

分别从蛇类药材和冷冻保存的新鲜蛇类肌肉标本中提取DNA,经PCR扩增出12种蛇共25个样品的Cyt b基因片段,并用银染测序的方法对DNA序列进行了分析.在此基础上用MEGA软件重建的系统发生树表明,研究的11种游蛇科蛇类可以分为3组:第一组为赤链蛇和水赤链游蛇,第二组为乌梢蛇和灰鼠蛇,第三组为锦蛇属的蛇,它们与第二组较近.锦蛇属是一高度分化的属,该组至少可分为两类,一类包括百花锦蛇和黑眉锦蛇;另一类包括玉斑锦蛇、棕黑锦蛇、红点锦蛇、王锦蛇和双斑锦蛇.后一类还可进一步分为3个亚组,玉斑锦蛇和棕黑锦蛇为第一亚组,红点锦蛇单独为第二亚组,王锦蛇和双斑锦蛇为第三亚组.本研究结果还表明,多年保存的陈旧药材标本可以用DNA序列分析的方法对其进行分子系统演化关系的研究.     

Genetic Identification of Spirometra decipiens Plerocercoids in Terrestrial Snakes from Korea and China

Human sparganosis is a zoonotic disease caused by infection with larval forms (procercoid/plerocercoid) of Spirometra spp. The purpose of this study was to identify Spirometra spp. of infected snakes using a multiplex PCR assay and phylogenetic analysis of mitochondrial DNA sequence data from the spargana of terrestrial snakes obtained from Korea and China. A total of 283 snakes were obtained that included 4 species of Colubridae comprising Rhabdophis tigrinus tigrinus (n=150), Dinodon rufozonatum rufozonatum (n=64), Elaphe davidi (n=2), and Elaphe schrenkii (n=7), and 1 species of Viperidae, Agkistrodon saxatilis (n=60). The snakes were collected from the provinces of Chungbuk, Chungnam, and Gyeongbuk in Korea (n=161), and from China (n=122). The overall infection rate with spargana was 83% (235/283). The highest was recorded for D. rufozonatum rufozonatum (100%), followed by A. saxatilis (85%) and R. tigrinus tigrinus (80%), with a negative result for E. davidi (0%) and E. schrenkii (0%). The sequence identities between the spargana from snakes (n=50) and Spirometra erinaceieuropaei ({"type":"entrez-nucleotide","attrs":{"text":"KJ599680","term_id":"646228164","term_text":"KJ599680"}}KJ599680) or S. decipiens ({"type":"entrez-nucleotide","attrs":{"text":"KJ599679","term_id":"646228151","term_text":"KJ599679"}}KJ599679) control specimens were 90.8% and 99.2%, respectively. Pairwise genetic distances between spargana (n=50) and S. decipiens ranged from 0.0080 to 0.0107, while those between spargana and S. erinaceieuropaei ranged from 0.1070 to 0.1096. In this study, all of the 904 spargana analyzed were identified as S. decipiens either by a multiplex PCR assay (n=854) or mitochondrial cox1 sequence analysis (n=50).     

缺抗蛇毒血清银环蛇伤的临床观察

目的探讨临床上缺乏抗银环蛇蛇毒血清治疗银环蛇咬伤的临床效果。方法将我院急诊科2011年3月~2013年10月收治的银环蛇咬伤患者9例与同期67例应用抗银环蛇毒血清治疗的病例进行比较,观察两组患者住院期间的临床变化与转归。结果经机械通气支持、胆碱酯酶抑制剂联合抗胆碱药物,以及肺部抗感染治疗,9例未经抗银环蛇毒血清治疗的患者均成活,但较使用抗银环蛇毒血清的患者住院时间延长、费用明显增加。结论重症银环蛇伤患者在缺乏抗银环蛇毒血清治疗时,应用机械呼吸支持,胆碱酯酶抑制剂联合抗胆碱药物,以及积极抗感染治疗也能救治患者生命。     

浙江省磐安县1973年—1985年蛇伤流行病学研究

对磐安县１９７３年￣１９８５年的蛇伤流行病学进行了回顾性调查。普查了全国２个国营林场、１个农场、３５６个行政村屯。１３年累计人口２４１２８８４人,其中蛇伤数为７３５人次（男５４６人次,女１８９人次）,死亡１０人。年平均发病数５６．５４人,发病率为２９．７／万,年均病死０．７７人,病死率为１．７７％,死亡率为０．０３７／万。各种致伤蛇种数高低依次是蛇蛇、竹叶青蛇、五步蛇、眼镜蛇、银环蛇、烙铁头蛇等。     

A comparative study of the biological properties of krait (genus Bungarus) venoms

1. The intravenous median lethal doses (LD50), protease, phosphodiesterase, alkaline phosphomonoesterase, L-amino acid oxidase, acetylcholinesterase, phospholipase A, 5'-nucleotidase, hyauronidase and anticoagulant activities of fourteen samples of venoms from the four common species of krait (Bungarus caeruleus, Bungarus candidus, Bungarus multicinctus and Bungarus fasciatus) were examined. 2. The results indicate that even though there are individual variations in the biological properties of the krait venoms, interspecific differences in the properties can be used for differentiation of the venoms from the four species of Bungarus. Particularly useful for this purpose are the LD50's and the contents of 5'-nucleotidase and hyaluronidase of the venoms.     

壶瓶山发现湖南蛇类新纪录二种——双全白环蛇和颈槽蛇

2002年6月至2008年8月,作者对湖南壶瓶山国家级自然保护区两栖爬行动物资源进行了10次实地调查与监测,共采集到蛇类32种.其中双全白环蛇(bycodon fasciatus)和颈槽蛇(Rhabdophisnuchalis)为湖南省蛇类新纪录.     

Declining snake population—why and how: a case study in the Mangrove Swamps of Sundarban, India

Species diversity of reptiles is much higher in Sundarban than in other mangrove ecosystems in India. Presently snakes are declining in Sundarban due to habitat loss caused by tremendous population pressure. Besides, irresponsible killing to avoid snakebite both from poisonous and nonpoisonous varieties is another reason for the gradual declining snake population. According to an intensive household survey in 35 villages in Sundarban, about 22 species (five poisonous and 17 non-poisonous) of snakes have been identified and there appears to have been a general decline in densities. On the contrary, a large number of people are bitten and die from snakebite every year with 0.57 and 0.34 vulnerability and mortality rate per 10,000 people, respectively. The two most commonly observed poisonous species are the common krait, Bungarus caeruleus (51 %) and common cobra, Naja naja (40 %), and that of non-poisonous varieties are the Ptyas mucosus (41 %), Typhlina bramina (34 %), Xenochrophis piscator (12 %), and Amphiesma stolata (10 %). Apart from killing of snakes out of fear; habitat loss, unscientific handling of snakes by snake catchers and charmers, and netting by fisherman contributes to snake mortality to a large extent; 72 % killed snakes are of poisonous varieties, 60 % of which are B. caeruleus, the most venomous snake in Sundarban. This paper is an attempt to highlight some of the important conservation efforts like the introduction of snake firms, alternative employment channels for the snake charmers and catchers, and mass awareness campaign through panchayet (village level governing body) and local NGOs.     

The complete amino-acid sequence of a basic phospholipase A2 in the venom of Bungarus fasciatus

A basic phospholipase A2 (PLA2) was isolated and purified from the venom of Bungarus fasciatus. Four kinds of enzymes, lysyl endopeptidase, endoproteinase Asp-N, endoproteinase Glu-C and trypsin, were employed to elucidate the complete primary structure by means of gas-phase sequencing. The amino-acid sequence reveals 118 amino-acid residues containing seven pairs of half-cystine. It has 78% and 61% structural identities with PLA2 from Bungarus multicinctus and Naja melanoleuca DE-II, respectively.     

浙江舟山五种卵生游蛇科动物个体大小、窝卵数和卵大小之间的关系

阐明五种游蛇科动物雌体大小、窝卵数和卵大小之间的关系和雌性繁殖特征的种间差异。５种蛇均产单窝卵,产卵高峰期为６月下旬至７月,窝卵数与雌体大小（ＳＶＬ）呈显著的正相关,相对窝卵重与雌体ＳＶＬ无关,卵理与窝卵数无关。灰鼠蛇卵重与雌体ＳＶＬ呈正相关,赤链蛇、王锦蛇、黑眉锦蛇和乌梢蛇的卵重与雌体ＳＶＬ无关,黑眉锦蛇卵长径与窝卵娄呈负相关,其余４种蛇卵长径与窝卵数无关。５种蛇卵长径与短径无关。黑眉锦蛇卵短径     

Authentication of snakes used in Chinese medicine by sequence characterized amplified region (SCAR)

Random amplified polymorphic DNA fingerprints characteristic of thethree snakes Zaocys dhumnades, Agkistrodonacutus and Bungarus multicinctus multicinctuswere generated using primer OPF-14. Z. dhumnades is anendangered species included in the Convention on International Trade inEndangered Species of Wild Fauna and Flora, and A. acutus,B. multicinctus multicinctus and Z. dhumnades are listed inthe Chinese Pharmacopoeia. The species-specific polymorphic bands Aa specific toA. acutus, Bmm specific to B. multicinctusmulticinctus and Zd specific to Z. dhumnadeswere identified and the sequences of these bands were used to design polymerasechain reaction primers for sequence characterized amplified region (SCAR)analysis of the three snakes. A multiplex SCAR analysis was established toauthenticate the snakes used in Chinese medicine reliably and efficiently.     

Snake Species Discrimination by Wild Bonnet Macaques (Macaca radiata)

Wild bonnet macaques (Macaca radiata) were studied in southern India to assess their ability to discriminate non‐venomous, venomous and predatory snakes. Realistic snake models were presented to eight troops of bonnet macaques at feeding stations and their behavior was video‐recorded 3 min before and 3 min after snake exposure. Snakes presented were: (1) venomous Indian cobra (Naja naja) displaying an open hood with ‘eyespots’; (2) venomous common Indian krait (Bungarus caeruleus); (3) non‐venomous green keelback (Macropisthodan plumbicolor); (4) non‐venomous rat snake (Ptyas mucosus); and (5) Indian python (Python molurus) which preys on macaques. Latencies to detect and react to the snakes were evaluated to determine initial responsiveness. Longer‐term assessment was measured as the percentage of time individuals looked at the snakes and monitored the activity of nearby individuals before and after snake detection. All snake models engendered caution and maintenance of a safe distance. Alarm calling occurred only during python presentations. The cobra engendered a startle response or running in the largest percentage of individuals after its detection, whereas the rat snake and python elicited bipedal standing or ambulating to monitor the snakes. We also examined the influence of age on snake recognition. Juveniles and subadults looked at the cobra, krait, and python for a larger percentage of time than adults did; albeit, adults looked at the python substantially longer than at the other snakes. Age differences in behavior suggest that, with the exception of the python, repeated experience with snakes in the wild moderates excitability, consistent with the likely threat of envenomation.     

Sequence variability in three mitochondrial DNA regions of Spirometra erinaceieuropaei spargana of human and animal health significance

Sequence variability in three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 3 (cox3), NADH dehydrogenase subunits 1 and 4 (nad1 and nad4) in Spirometra erinaceieuropaei spargana from different geographical regions in China was examined. A portion of each of the cox3 (pcox3), nad1 (pnad1) and nad4 genes (pnad4) were amplified separately from individual S. erinaceieuropaei spargana by polymerase chain reaction (PCR). Representative amplicons were subjected to sequencing in order to estimate sequence variability. The sequences of pcox3, pnad1 and pnad4 were 541, 607 and 847?bp in length, respectively. The A+T contents of the sequences were 68.39-68.76% (pcox3), 63.76-64.91% (pnad1) and 67.18-67.77% (pnad4), respectively, while the intra-specific sequence variations within each of the S. erinaceieuropaei spargana were 0-1.5% for pcox3, 0-2.8% for pnad1 and 0-2.7% for pnad4. Phylogenetic analysis using neighbour joining (NJ), maximum likelihood (ML) and maximum parsimony (MP) methods, indicated that all the spargana isolates in Hunan Province represented S. erinaceieuropaei. These findings demonstrated clearly the usefulness of the three mtDNA sequences for population genetics studies of S. erinaceieuropaei spargana of human and animal health significance.     

Influences of area, isolation and habitat features on distribution of snakes in Mediterranean fragmented woodlands

The effects of isolation-related and vegetational parameters on presence and relative abundance of snakes in patchy forested fragments of Mediterranean central Italy are studied. The most abundant species was Coluber viridiflavus (accounting for 47.7% of the total snake sample observed) followed by Vipera aspis (22%), Elaphe longissima (21.5%), Natrix natrix (7.7%), and Coronella austriaca (1.1%). There was a clear trend for bigger species to be less distributed among the various forest fragments than the smaller species. Presence of Coluber viridiflavus, Coronella austriaca and Natrix natrix was not influenced by woodland area, whereas that of Vipera aspis and Elaphe longissima was positively influenced by woodland area. Woodland isolation parameters did not influence the presence of Coluber viridiflavus, Coronella austriaca and Natrix natrix, but of Vipera aspis and Elaphe longissima. Discriminant stepwise analysis suggested that specific environmental features influenced the occurrence and abundance of the various snake species, Vipera aspis being the taxon more affected by isolation-related parameters. Some conservation implications of our observations are also discussed.     

Primary structure of an inactive mutant of phospholipase A2 in the venom of Bungarus fasciatus (banded krait).

From the acidic components of Bungarus fasciatus venom, a very small amount (0.16%) of a novel phospholipase A2 was obtained. Both neurotoxicity and enzyme activity were found to be lacking. Amino acid sequence study showed that it has a normal backbone of group I snake venom phospholipase A2 with 118 amino acid residues. The lack of enzyme activity was attributed to its mutation of the indispensable Asp residue to an Ala residue, i.e., the usual His-Asp47 turned out to be His-Ala47. This is the eighth isoform of phospholipase A2 found from the venom of Bungarus fasciatus. Examination of structural homology with three other isoforms revealed 66% similarity at most.     

cDNA sequence and molecular modeling of a nerve growth factor from Bothrops jararacussu venomous gland

The complete nucleotide sequence of a nerve growth factor precursor from Bothrops jararacussu snake (Bj-NGF) was determined by DNA sequencing of a clone from cDNA library prepared from the poly(A) + RNA of the venom gland of B. jararacussu. cDNA encoding Bj-NGF precursor contained 723 bp in length, which encoded a prepro-NGF molecule with 241 amino acid residues. The mature Bj-NGF molecule was composed of 118 amino acid residues with theoretical pI and molecular weight of 8.31 and 13,537, respectively. Its amino acid sequence showed 97%, 96%, 93%, 86%, 78%, 74%, 76%, 76% and 55% sequential similarities with NGFs from Crotalus durissus terrificus, Agkistrodon halys pallas, Daboia (Vipera) russelli russelli, Bungarus multicinctus, Naja sp., mouse, human, bovine and cat, respectively. Phylogenetic analyses based on the amino acid sequences of 15 NGFs separate the Elapidae family (Naja and Bungarus) from those Crotalidae snakes (Bothrops, Crotalus and Agkistrodon). The three-dimensional structure of mature Bj-NGF was modeled based on the crystal structure of the human NGF. The model reveals that the core of NGF, formed by a pair of beta-sheets, is highly conserved and the major mutations are both at the three beta-hairpin loops and at the reverse turn.     

蛇毒清合剂治疗金环蛇咬伤的疗效观察

目的观察蛇毒清合剂对金环蛇咬伤的临床疗效。方法将71例病人分为治疗组35例,对照组36例。两组均给予常规治疗,治疗组加服蛇毒清合剂,并比较两组患者伤后48h的血清酶学三项（CK、LDH、AST）及治疗5天后的疗效、治疗结束时的疗程及呼吸衰竭发生率。结果两组患者临床疗效比较有显著性差异,治疗组优于对照组（P〈0．05）,酶学三项和疗程、呼吸衰竭发生率均有显著性差异（P〈0．05）。结论蛇毒清合剂对金环蛇咬伤有显著疗效。