我国蛇类常见寄生虫及其对人类健康的影响

目的研究我国蛇类常见寄生虫对人类健康的影响。方法收集和分析国内外蛇类寄生虫对人类健康影响的研究资料。结果我国常见危害人类健康的蛇源寄生虫有6种,即舌形虫、曼氏迭宫绦虫、隐孢子虫、颚口线虫、广州管圆线虫、线中殖孔绦虫,均为人兽共患性寄生虫,对人类健康造成危害。结论通过加强科普宣传,加强科研工作,加强执法监管,规范蛇类养殖产业,可有效防控蛇类常见寄生虫对人类健康的威胁。     

试验感染水牛体内枯氏住肉孢子虫内生发育的超微结构

通过人工感染试验在电镜下对一种住肉孢子虫在水牛体内的发育进行了研究。结果揭示在血管内皮细胞中的裂殖生殖和在肌细胞内的包囊发育过程与枯氏住肉孢子虫的一致,裂殖体、裂殖子、包囊壁和囊内母细胞、缓殖子的超微结构亦与枯氏住肉孢子虫的相同。首次通过试验感染的发育史研究证明,流行于我国湖南水牛的一种小包囊属于枯氏住肉孢子虫,水牛是黄牛枯氏住肉孢子虫的新宿主。     

我国主要海域海水养殖碳汇能力评估及其影响效应——基于我国9个沿海省份面板数据

随着海水养殖业的碳汇功能逐渐被认识和肯定,海水养殖不再单是一项经济活动,而是对环境具有正向影响的碳汇生态活动。以我国沿海9个省份为例,选取海水养殖业碳汇主要贡献的贝类和藻类海产品,并按照各自的碳汇方式对我国沿海地区2008—2015年海水养殖碳汇能力测算,进一步将9个沿海省份按照主要海域划分为渤海、黄海、东海、南海,利用LMDI模型从海水养殖的结构效应和规模效应角度分析碳汇能力的区域差异和主要影响因素。研究结果显示,黄海沿岸海水养殖碳汇能力最强,南海沿岸海水养殖的碳汇转化比例最高,规模效应与我国沿海地区海水养殖碳汇能力始终呈正相关,结构效应的作用显著但不稳定。基于上述结论,我国沿海地区碳汇养殖业应首先提升碳汇养殖技术、稳定海水养殖产量,其次注重优化养殖结构,对碳汇潜力巨大的贝类多加关注。     

粘孢子虫分类学研究进展

粘孢子虫是变温动物体内常见的寄生虫,绝大部分寄生于鱼类。近年来,粘孢子虫引发的病害日益严重,但其基础研究尚处于初级阶段,尤其在分类地位和分类系统方面还存在许多争议。分子生物学方法的快速发展与应用,使粘孢子虫的分类学研究取得了长足进步。该文从粘孢子虫分类地位与起源、分类系统和分类学方法等三个方面对国内外粘孢子虫分类学研究的现状进行了综述,同时就目前粘孢子虫在分类地位和起源中存在的分歧观点以及分类方法的发展趋势等做了讨论,以期为鱼类粘孢子虫病的研究奠定基础。     

青岛近岸海水及几种海产品中副溶血弧菌(Vibrio parahaemolyticus)生境的调查

副溶血弧菌是水产食品中的一种重要食物中毒菌,它在沿岸海水和海产动物体上都有广泛的分布,所以它又是海产动物的一种条件性致病菌。本文报告了该菌在青岛近岸海水和几种常见的水产食品上污染了该菌的情况,在检测方法上也进一步修定了一种选择性较强的培养基(SAC)。调查发现青岛近岸海水中副溶血弧菌的出现率为68～463CFU/100ml,几种常见海产食品中平均含量值为745～6900CFU/100克,其中贝类含量最高达6900/100克。采用常用的选择培养基“TCBS”等与修定的“SAC”对该菌进行检测比较,结果表明“SAC”法较“TCBS”法检出的阳性率和灵敏性均较高,两种方法X~2=4.28,P>0.05,效果有显著的意义。     

蚯蚓的寄生虫

对于蚯蚓寄生虫的调查研究工作,在我国起步较晚,并仅限于猪肺线虫的研究,在国外研究得较多,在５０年代以后,有关的报道更多,蚯蚓的寄生虫,最为人知的为原生动物中的簇虫类,纤毛虫类和其它的孢子虫。其次是旋毛虫类、扁虫类、线虫类、轮虫类、寡毛类和双翅目昆虫的幼虫等。     

鮸鱼弧菌病病原菌(哈维氏弧菌)的分离与鉴定

【目的】2009年春季,浙江省舟山地区养殖鮸鱼暴发弧菌病。症状主要表现为体表病灶部位出血、肌肉溃烂、内脏器官有白斑等。【方法】从病鱼体表溃疡部位及内脏分离出优势菌株090212,经人工感染证实该菌即为致病菌。通过API系统和菌体常规形态特征、培养特性和生理生化反应指标测定以及16S rRNA测序分析等综合鉴定,【结果】确认090212为哈维氏弧菌(Vibrio harveyi),该菌为革兰氏阴性,菌体呈短杆状,极生单鞭毛。该菌对氟苯尼考、四环素等5种抗生素敏感。【结论】哈维氏弧菌是海水养殖鱼类的常见致病菌,但作为养殖鮸鱼的病原菌尚属首次报道,将对鮸鱼的病害防治和健康养殖具有重要的指导意义。     

尼氏单极虫(粘体门,双壳目)重描述及基于18S rDNA系统地位分析

采用常规细胞学方法与18S rDNA序列比较的方法,对采自于重庆市嘉陵江段的粘孢子虫,尼氏单极虫Thelohanellus nikolskii进行了形态学和分子生物学的研究,此为首次在我国长江流域检获尼氏单极虫。本文在补充了尼氏单极虫中国种群18S rDNA序列信息的分析、比较基础上,探讨了该虫与相邻种属的系统关系,佐证了其分类学位置。     

多倍体经济贝类的育种研究现状

海水经济贝类在海水养殖中占有重要的地位,开展贝类育种、育苗对提高重要经济贝类的产量和质量意义重大。多倍体育种作为贝类育种的方法之一,研究者希望通过这种方法,诱导出个体大、生长快、肉质好的三倍体贝类,以提高群体生长速度,改善经济性状,增强养殖生物的抗逆性。对多倍体贝类的亲贝培育、育种原理、诱导方法、检测技术,以及生物学性状作了综述,最后探讨了多倍体研究中存在的主要问题。     

厦门海域贝类养殖环境中有机氯农药的积累和降解

利用气相色谱法(GC-ECD)对厦门海域贝类养殖环境(海水、底质和养殖贝类)中有机氯农药六六六(HCH)和滴滴涕(DDT)的含量进行了调查分析,初步探讨了贝类养殖环境中HCH和DDT的积累和降解规律.结果表明:厦门海域不同种贝类养殖环境中HCH和DDT的积累和降解存在明显差异,主要与各种贝类的栖息环境和生理生活习性有关.贝类养殖环境中处于积累状态(Rx＞1)的主要为p-HCH、δ-HCH和γ-HCH;处于降解状态(Rx＜1)的主要为α-HCH.α-HCH/γ-HCH的比值≤1.0,贝类养殖环境中的HCH来源于工业品HCH和林丹,大部分HCH为长时间残留,但尚有少量林丹输入.贝类养殖区海水中DDT主要为好氧降解,底质中主要为厌氧降解.海水中的降解产物主要为DDE,(DDD+ DDE )/DDTs(p,p-DDE+p,p-DDD+o,p-DDT+p,p-DDT)的比值＜0.5;底质和养殖贝类体中的降解产物主要为DDD,(DDD+DDE)/DDTs的比值＞0.5,贝类养殖区底质和养殖贝类体中的DDT大部分已降解为DDD和DDE,海水中尚有少量新的DDT输入.贝类养殖环境中HCH异构体降解率的高低顺序存在一定差异,其在贝类养殖生态系中的迁移、转化过程发生了构象变化.     

Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes

Haplosporidian parasites infect various invertebrate hosts including some commercially important shellfish. Haplosporidium nelsoni (along with Perkinsus marinus) has severely affected Eastern oyster production on the eastern seaboard of the United States and flat oyster production in Europe has been severely impacted by Bonamia ostreae. These parasites are also often present at a very low prevalence and there are a variety of morphologically similar species that can be difficult to differentiate during cytological or histological diagnosis hence the need to develop specific tests. Recently, a Minchinia sp. was described affecting rock oysters (Saccostrea cuccullata) in north Western Australia. In this study, two in situ hybridisation (ISH) assays and a PCR assay have been developed and optimised for use in investigating these parasites. The first ISH assay used a 166bp polynucleotide probe while the second used a 30bp oligonucleotide probe. The specificity of each ISH assay was assessed by applying each probe to a variety of haplosporidian (5), a paramyxian (1) or ciliophora (1) parasites. The polynucleotide probe produced strong hybridisation signals against all of the haplosporidian parasites tested (Minchinia sp., Minchinia teredinis, Bonamia roughleyi, H. nelsoni and Haplosporidium costale) while the oligonucleotide probe recognised only the Minchinia sp. Both probes failed to detect the paramyxian (Marteilia sp.) or the Rhynchodid-like ciliate. The PCR assay amplifies a 220bp region and detected Minchinia sp. DNA from 50ng of genomic DNA extracted from the tissues of infected oysters and 10fg of amplified Minchinia sp. DNA. The assay did not react to oysters infected with H. nelsoni or H. costale. The ability of the PCR and oligonucleotide ISH assay to diagnose Minchinia sp. infected oysters was compared to histological examination from a sample of 56 oysters. The PCR assay revealed 26 infections while histological examination detected 14 infections. The oligonucleotide ISH assay detected 29 infections. The oligonucleotide ISH and PCR assays were found to be significantly more sensitive than histology for detecting the parasite.     

Spore ornamentation of Haplosporidium nelsoni and Haplosporidium costale (Haplosporidia), and incongruence of molecular phylogeny and spore ornamentation in the Haplosporidia

Spore ornamentation of Haplosporidium nelsoni and Haplosporidium costale was determined by scanning electron microscopy. For H. nelsoni, the spore surface was covered with individual ribbons that were tightly bound together and occurred as a single sheet. In some spores, this layer was overlaid with a network of branching fibers, about 0.05 microm in diameter, which often was dislodged from the spore at the aboral pole. For H. costale, ornamentation consisted of a sparse network of branching fibers on the spore surface. Molecular phylogenetic analysis of the phylum Haplosporidia revealed that Urosporidium, Bonamia, and Minchinia were monophyletic but that Haplosporidium was paraphyletic. All species of Minchinia have ornamentation composed of epispore cytoplasm, supporting the monophyly of this genus. The presence of spores with a hinged operculum and spore wall-derived ornamentation in Bonamia perspora confounds the distinction between Bonamia and Haplosporidium. Species with ornamentation composed of outer spore wall material and attached to the spore wall do not form a monophyletic group in the molecular phylogenetic analysis. These results suggest that the widely accepted practice of assigning all species with spore wall-derived ornamentation to Haplospordium cannot be supported and that additional genera are needed in which to place some species presently assigned to Haplosporidium.     

Prevalence of Perkinsus marinus (dermo), Haplosporidium nelsoni (MSX), and QPX in bivalves of Delaware's inland bays and quantitative, high-throughput diagnosis of dermo by QPCR

Restoration of oyster reef habitat in the Inland Bays of Delaware was accompanied by an effort to detect and determine relative abundance of the bivalve pathogens Perkinsus marinus, Haplosporidium nelsoni, and QPX. Both the oyster Crassostrea virginica and the clam Mercenaria mercenaria were sampled from the bays. In addition, oysters were deployed at eight sites around the bays as sentinels for the three parasites. Perkinsus marinus prevalence was measured with a real-time, quantitative polymerase chain reaction (PCR) methodology that enabled high-throughput detection of as few as 31 copies of the ribosomal non-transcribed spacer region in 500 ng oyster DNA. The other pathogens were assayed using PCR with species-specific primers. Perkinsus marinus was identified in Indian River Bay at moderate prevalence ( approximately 40%) in both an artificial reef and a wild oyster population whereas sentinel oysters were PCR-negative after 3-months exposure during summer and early fall. Haplosporidium nelsoni was restricted to one oyster deployed in Little Assawoman Bay. QPX and P. marinus were not detected among wild clams. While oysters in these bays have historically been under the greatest threat by MSX, it is apparent that P. marinus currently poses a greater threat to recovery of oyster aquaculture in Delaware's Inland Bays.     

Spore ornamentation of Minchinia occulta n. sp. (Haplosporidia) in rock oysters Saccostrea cuccullata (Born, 1778)

A Minchinia sp. (Haplosporidia: Haplosporidiidae) parasite was identified infecting rock oysters and morphologically described by Hine and Thorne (2002) using light microscopy and transmission electron microscopy (TEM). The parasite was associated with up to 80% mortality in the host species and it is suspected that the parasite would be a major impediment to the development of a tropical rock oyster aquaculture industry in northern Western Australia. However, attempts to identify the parasite following the development of a specific probe for Haplosporidium nelsoni were unsuccessful. The SSU region of the parasite's rRNA gene was later characterized in our laboratory and an in situ hybridization assay for the parasite was developed. This study names the parasite as Minchinia occulta n sp. and morphologically describes the parasite using histology, scanning electron microscopy and transmission electron microscopy. The non-spore stages were unusual in that they consisted primarily of uninucleate stages reminiscent of Bonamia spp. The parasite's spores were ovoid to circular shaped and measured 4.5 microm-5.0 microm x 3.5-4.1 microm in size. The nucleus of the sporoplasm measured 1.5-2.3 microm and was centrally located. The spores were covered in a branching network of microtubule-like structures that may degrade as the spore matures.     

牡蛎单孢子虫环介导等温扩增可视化检测方法的建立

目的:建立基于环介导等温扩增(LAMP)技术的单孢子虫可视化检测方法。方法:根据尼氏单孢子虫的小亚基核糖体RNA保守序列,设计一套特异性LAMP引物,对反应条件如温度和试剂浓度进行优化,建立检测牡蛎单孢子虫的LAMP方法。结果:所建立的方法的敏感性可达1 fg,是常规PCR方法的100倍;全部反应可在1 h内完成;可通过肉眼观察颜色,直接判定结果;对其他牡蛎常见病原体的检测结果均为阴性。结论:建立的LAMP方法简便、快速、灵敏、特异,可用于牡蛎单孢子虫感染的快速检测。     

Widespread survey finds no evidence of Haplosporidium nelsoni (MSX) in Gulf of Mexico oysters

The advent of molecular detection assays has provided a set of very sensitive tools for the detection of pathogens in marine organisms, but it has also raised problems of how to interpret positive signals that are not accompanied by visual confirmation. PCR-positive results have recently been reported for Haplosporidium nelsoni (MSX), a pathogen of the oyster Crassostrea virginica in 31 of 40 oysters from 6 sites in the Gulf of Mexico and the Caribbean Sea. Histological confirmation of the PCR results was not undertaken, and no haplosporidian has been reported from the numerous histological studies and surveys of oysters in the region. To further investigate the possibility that H. nelsoni is present in this region, we sampled 210 oysters from 40 sites around the Gulf of Mexico and Puerto Rico using PCR and 180 of these using tissue-section histology also. None of the oysters showed evidence of H. nelsoni by PCR or of any haplosporidian by histology. We cannot, therefore, confirm that H. nelsoni is present and widespread in the Gulf of Mexico and the Caribbean Sea. Our results do not prove that H. nelsoni is absent from the region, but taken together with results from previous histological surveys, they suggest that for the purposes of controlling oyster importation, the region should continue to be considered free of the parasite.     

中国近海入侵贝类及其影响

海洋贝类种类繁多,是近海底栖生态系统的优势种,在生态系统物质循环和能量流动中起着极其重要的作用;但因频繁的人类活动导致的贝类入侵问题,往往使该过程不能正常运转。大量研究表明,入侵贝类会抑制原著贝类或其他生物的生长及拓殖,形成单一优势群落,破坏当地生态系统的平衡,也会对海洋渔业生产和近海工程及作业等造成不可预测的危害。目前,欧美等国家针对该问题已开展了大量工作,十分重视入侵贝类的生物学、生态学与行为学特征及特性的研究,并根据其入侵途径与方式的不同开展了检测、监测、防控及管理工作。但在我国,入侵贝类还未引起相关部门及研究机构的足够重视。本文主要简述了我国近海的3种入侵贝类沙筛贝、指甲履螺和地中海贻贝的生物学、生态学及行为学等特征及其对当地生态系统的影响;同时,根据其特性及入侵过程控制的难易,建议在完善我国入侵贝类数据库的基础上加强早期预警与检测,以及安全防控和管理等工作。     

Immunoassay Comparison of Haplosporidan Spores from Teredo navalis and Haplosporidium nelsoni Spores from Crassostrea virginica

Spores of a haplosporidan infecting Teredo navalis Linnaeus have been described as morphologically indistinguishable from spores of Haplosporidium nelsoni . To test the hypothesis that these organisms are conspecific, a colloidal gold immunoassay was used to compare antigenic characteristics of the spores from both hosts. Rabbit antibody to formalin-fixed spores from T. navalis was tested against paraffin sections of Crassostrea virginica infected with spores of H. nelsoni and against paraffin sections of infected 7". navalis . Application of primary antibody was followed by addition of affinity purified goat anti-rabbit IgG coaled on 5-nm colloidal gold particles. The reaction was enhanced by precipitation of metallic silver; a positive reaction appeared as a dark brown to black signal at the site of each antigen-antibody complex. Haplosporidium nelsoni spores did not react when assayed with the antibody made to spores from T. navalis . Spores from infected T. navalis tissue reacted positively with rabbit antibody. This result indicates that the spores from the 2 hosts are antigenicaliy distinct and suggests that they are different species.     

Efficacy of different administration routes for vaccination against Vibrio anguillarum in Atlantic halibut (Hippoglossus hippoglossus L.)

Atlantic halibut (Hippoglossus hippoglossus L.) is a potentially important new species to cold-water aquaculture. Development of a viable industrial farming technique has been hampered by continued pathogen problems within the rearing cycle and there are several reports that indicated how susceptible juvenile halibut are to bacterial and viral diseases. Interest has been expressed, within the industry, over the possibility of vaccinating suitably sized animals to protect against the more common aquaculture pathogens. Vibrio spp. are of particular concern due to their ubiquitous nature and the relatively frequent occurrence of these pathogens within marine aquaculture. We have previously investigated the susceptibility of Atlantic halibut to infection by Vibrio anguillarum and the efficacy of intraperitoneal injected delivery of a commercial vaccine in protecting against the disease. Given the very high rate of protection offered by immunisation we wanted to investigate the effect of alternate routes of administration on the efficacy of the vaccine.