鱼类三代虫分子生物学的研究进展

三代虫是一类常见的鱼类体外寄生虫,绝大多数种类寄生于鱼类的体表和鳃,大量寄生可导致苗种甚至成鱼大批死亡,或引起其他病原感染而致继发性鱼病,造成严重的经济损失.本文从三代虫目前的分类学概况、DNA分类学和条形码技术、核糖体基因及线粒体基因的研究等方面概述了鱼类三代虫分子生物学的研究现状,为三代虫及扁形动物的分子生物学研究及其应用提供有用的资料.     

寄生于洪湖经济鱼类鳃上的单殖吸虫包括二新种的描述

本文报告寄生于洪湖经济鱼类鳃上的单殖吸虫共60种,包括描述指环虫和三代虫各一新种,并有若干寄主新记录及长江流域新记录。     

额尔齐斯河几种鱼类寄生三代虫种类鉴定及系统发育

对采自额尔齐斯河的银鲫(Carassius auratus gibelio Bloch)、东方欧鳊(Abramis brama orientalis Berg)、黏鲈(Gymnocephalus cernua Linnaeus)、金鳟(Oncorhynchus mykiss Walbaum)感染的三代虫(Gyrodactylus sp.)进行了研究, 通过几丁质结构的形态测量与比较, 初步鉴定寄生于银鲫的为细锚三代(G. sprostonae), 寄生东方欧鳊的为秀丽三代虫(G. elegans), 寄生黏鲈的为普氏三代虫(G. prostate), 寄生金鳟的为细鳞鲑三代虫(G. brachymystacis)。同时将测定的三代虫的ITS序列, 与GenBank上三代虫序列进行比对分析, 发现上述4种三代虫分别与细锚三代虫、秀丽三代虫、细鳞鲑三代虫、普氏三代虫的ITS同源性都在99.3%以上, 进一步验证了形态学的鉴定结果。系统发育分析显示细鳞鲑三代虫和细锚三代虫位于G. (Limnonephrotus)亚属的一枝, 秀丽三代虫和普氏三代虫位于G. (Gyrodactylus)亚属的一枝, 而此两亚属亲缘关系较远。     

鱼类三代虫的寄主特异性分析

分析了迄今为止世界范围三代虫的寄主情况。三代虫具有很强的寄主特异性,并且在种、属及科等水平上的特异性均有较明显差异;分析同一种寄主上所寄生的三代虫种类之间的关系,显示大部分寄主只寄生一种三代虫,体现出了三代虫很强的寄主特异性。     

金鱼寄生三代虫的形态学及基于rDNAits序列的分子鉴定

为查明我国金鱼三代虫的种类, 研究采用形态学与分子生物学相结合的方法, 对我国6个金鱼主要养殖区的三代虫进行了调查。通过形态测量分析, 发现了4种三代虫, 分别为: 小林三代虫（Gyrodactylus kobayashii）、G. gurleyi、G. longoacuminatus和三代虫未定种（Gyrodactylus sp.）。而通过核糖体rDNA its序列比对分析, 发现有2种三代虫分别与小林三代虫和G. longoacuminatus相似性在99.1%以上, 有1种与G. gurleyi的相似性在98.2%以上, 另外, 还有1种三代虫与现有序列的相似性都较低, 虽然它与小林三代虫的相似性最高, 也仅为91.0%, 暂为未定种。系统发育分析也显示, 金鱼寄生的3种三代虫分别与小林三代虫、G. gurleyi和G. longoacuminatus聚集在一起, 另外1种单独聚在一起, 这4种三代虫均隶属于G. （Limnonephrotus）亚属, 与G. （Gyrodactylus）亚属的秀丽三代虫（G. elegans）的亲缘关系很远。通过对三代虫样品的抽样调查发现, 小林三代虫存在于所有采样点, 在广州、武汉、郑州3个采样点为优势种, 所占的比率都在60%以上; G. gurleyi在海口、南京和上海为优势种, 所占的比率都在50%以上; G. longoacuminatus和三代虫未定种所占比率均较小。由此可见, 小林三代虫和G. gurleyi是我国金鱼常见的三代虫, G. gurleyi 和G. longoacuminatus是中国三代虫新记录种。       

秦岭山区虹鳟寄生三代虫一新纪录

记述了秦岭山区寄生于虹鳟的单殖亚纲一新纪录种：细鳞鲑三代虫Gyrodactylus brachymystacis Ergens,1978,比较了其与我国东北寄生于虹鳟的细鳞鱼三代虫Gyrodactylus lenoki Gussev,1953的区别。初步判断其来源可能一是通过水源来源于秦岭的细鳞鲑,另一种是通过早期虹鳟的引进带人的,其真实的途径及其对环境的风险性还有待于进一步研究。     

铜鱼寄生三代虫一新种的记述

本文报道我国长江中游铜鱼鳃部寄生的三代虫一新种,即:铜鱼三代虫(新种)Gyrodactylus coreiusisp.nov.,新种的形态特征及其与相近种之间的差别在文中进行了描述和比较。     

常用消毒剂次氯酸钠和二氧化氯对小林三代虫的杀灭效果

为了有效控制三代虫病, 实验以寄生于金鱼的小林三代虫(Gyrodactylus kobayashii)为动物模型, 研究了两种常用消毒剂次氯酸钠溶液(NaClO)和二氧化氯(ClO2)的杀虫效果。结果表明: 在离体(in vitro)条件下, 当NaClO的有效浓度0.2 mg/L或ClO2的有效浓度0.15 mg/L 时, 小林三代虫的平均存活时间均少于2h, 而对照组中小林三代虫的平均存活时间是20.8h。当ClO2的有效浓度0.15 mg/L时, 70%以上的虫体发黑, 而其他浓度处理组, 大部分虫体即使死亡, 虫体依然保持透明。在在体(in vivo)条件下, 当 NaClO的有效浓度0.2 mg/L或ClO2的有效浓度0.5 mg/L 时, 驱虫率都几乎达到100%, 并且驱虫率随着药物浓度的增加而提高,但当ClO2的有效浓度为0.6 mg/L时, 养殖水体出现了白色絮状物。在在体条件下, NaClO的驱虫效果好于ClO2。在金鱼的急性毒性实验中, NaClO和ClO2的安全浓度分别是0.18和0.48 mg/L, 仅稍低于其在在体条件下完全驱除小林三代虫的最小浓度(0.2、0.5 mg/L), 说明次氯酸钠溶液和二氧化氯在驱除三代虫时对金鱼不太安全, 因此, 在治疗金鱼的三代虫病时要慎使次氯酸钠溶液和二氧化氯。然而, 这两种消毒剂能否适用于其他鱼类三代虫病的治疗则有待进一步研究。     

鳑鮍亚科鱼类鳃部寄生的四种三代虫的记述

据文献记载 ,有关我国三代虫 (Ｇｙｒｏｄａｃｔｙｌｉｎａｅ)的研究始于 1 948年[1],迄今为止 ,已报道三代虫 42种[1—3]。作者在整理、鉴定三代虫标本时发现 ,寄生于两种鳃部的四种三代虫在我国均未见报道 ,其中有两新种 ,现记述如下。量度均以毫米为单位。模式标本保存在中国科学院水生生物研究所鱼病学研究室。1　东湖三代虫 ,新种Ｇｙｒｏｄａｃｔｙｌｕｓｄｏｎｇｈｕｅｎｓｉｓｓｐ .ｎｏｖ .(图 1 )宿主与寄生部位 :中华ＲｈｏｄｅｕｓｓｉｎｅｎｓｉｓＧ櫣ｎｔｈｅｒ的鳃。图 1　东湖三代虫Ｇ .ｄｏｎｇｈｕｅｎｓｉｓｓｐ…     

牡丹江鱼类单殖吸虫一新记录

近年来,三代虫在黑龙江省各虹鳟鱼场普遍发生,为害程度与日俱增。       

Effect of Australian tea tree oil on Gyrodactylus spp. infection of the three-spined stickleback Gasterosteus aculeatus

Gyrodactylus spp. infections of commercially farmed fishes are responsible for significant economic losses. Existing treatments have proved uneconomic, stressful to the fishes, and ecologically damaging. Essential oils are naturally occurring compounds that exhibit a wide range of anti-microbial and anti-fungal activities. This study explored the possibility of using Australian tea tree (Melaleuca alternifolia) oil (TTO) to treat Gyrodactylus spp. infection on the three-spined stickleback Gasterosteus aculeatus. In the presence of 0.01 % Tween 80 as an emulsifier, TTO treatments at concentrations between 3 and 30 ppmv (parts per million by volume) lowered the prevalence and significantly reduced the parasite burden of sticklebacks naturally infected with Gyrodactylus spp. In addition, Tween 80 alone exhibited parasiticidal activity against Gyrodactylus spp. These findings show the potential of TTO in combination with Tween 80 as an effective treatment of Gyrodactylus spp. infection of fishes.     

Two new species of Gyrodactylus von Nordmann, 1832 (Monogenea: Gyrodactylidae) parasitizing Girardinichthys multiradiatus (Cyprinodontiformes: Goodeidae), an endemic freshwater fish from central Mexico

Gyrodactylus mexicanus n. sp. and Gyrodactylus lamothei n. sp. are described from the fins and skin of Girardinichthys multiradiatus, an endemic freshwater fish from central Mexico. Gyrodactylus mexicanus is compared to other Gyrodactylus species that parasitize Fundulus spp., the phylogenetically closest group to the Goodeidae from North America. Gyrodactylus mexicanus is distinguished by having large anchors with well-developed superficial roots, enlarged hooks with a proximally disrupted shank (ligament), and a ventral bar with 2 poorly developed anterolateral projections and a small medial process. Gyrodactylus lamothei is distinguished from G. mexicanus and from other species of Gyrodactylus on the North American continent by having anchors with a sclerite on the superficial root and robust hooks with a straight shaft and a recurved point.     

Parasitic infections in freshwater ornamental fish in Sri Lanka

A total of 1520 ornamental fish of 13 species from 26 export farms in Sri Lanka were collected between October 1999 and March 2000 and examined for parasites. Fish species examined were guppy Poecilia reticulata, goldfish Carassius auratus, platy Xiphophorus maculatus, molly Poecilia sphenops, angel Pterophyllum scalare, swordtail Xiphophorus helleri, tetras Hyphessobrycon species, barbs Capeota and Puntius spp., gourami Colisa sp., carp Cyprinus carpio, fighters Betta spelendens and others (Brachydanio and Astronotus spp.). Nine species of monogenean trematodes (Dactylogyrus extensus, Dactylogyrus cf. extensus, D. vastator, Dactylogyrus cf. vastator Dactylogyrus spp., Gyrodactylus turnbulli, G. katherineri, Gyrodactylus cf. katherineri, Gyrodactylus spp.), 7 protozoan species (Trichodina nigra, Trichodina spp., Tetrahymena corlissi, T. pyriformis, Ichthyophthirius multifiliis, Ichthyobodo necator, Piscinoodinium spp.), 3 species of copepod arthropods (Lernaea cyprinacea, Ergasilus ceylonensis, Argulus foliaceus), 1 metacercarial stage of a digenean trematode (Centrocestus spp.) and 1 nematode (Capillaria spp.) were identified. Parasites were found in fish from 23 of the 26 farms with an overall prevalence of parasitism in 45.3% of fish. The variation in farm prevalence among different parasites was significant (p < 0.01). Fish infection rates with monogenean trematodes, protozoans, copepod crustaceans, digenean trematodes and nematodes were 28.3, 18.4, 4.8, 0.8 and 0.4%, respectively. In all, 50 out of 590 (50/590) guppies were infected with Tetrahymena, compared with 13/930 for all other species, which is a statistically significant result (p < 0.01). Similarly, 13/44 and 18/44 carp were infected with Argulus foliaceus and Lernaea cyprinacea, compared with 7/1476 and 15/1476, respectively, for all other species combined (p < 0.01). Capillaria spp. was found only in guppies (4/590) and angel fish (3/92) while Centrocestus spp. was found in goldfish (12/153) only.     

Experimental infections of the monogenean Gyrodactylus turnbulli indicate that it is not a strict specialist

Parasites represent a threat to endangered fish species, particularly when the parasite can host switch and the new host is vulnerable. If the parasite is highly host specific then successful host switching should be a rare occurrence; however, the host range of many parasites which are assumed to be specialists has never been tested. This includes the monogenean Gyrodactylus turnbulli, a well-studied ectoparasite found caudally on its known host, the guppy, Poecilia reticulata. In this study, we monitored parasite establishment and reproduction on a range of poeciliids and more distantly related fish. Individually maintained fish were experimentally infected with a single parasite and monitored daily to establish whether G. turnbulli could survive and reproduce on other fish species. Gyrodactylus turnbulli can infect a wider range of hosts than previously considered, highlighting the fact that host specificity can never be assumed unless experimentally tested. Our findings also have significant implications for parasite transmission to novel hosts and provide further insight into the evolutionary origins of this ubiquitous group of fish pathogens. Previous molecular evidence indicates that host switching is the key mechanism for speciation within the genus Gyrodactylus. Until recently, most Gyrodactylus spp. were assumed to be narrowly host specific. However, our findings suggest that even so-called specialist species, such as G. turnbulli, may represent a threat to vulnerable fish stocks. In view of the potential importance of host switching under artificial conditions, we propose to describe this as 'artificial ecological transfer' as opposed to 'natural ecological transfer', host switching under natural conditions.     

Gyrodactylus anarhichatis n. sp. and G. microanchoratus n. sp. (Monogenea: Gyrodactylidae) from the Atlantic wolf-fish Anarhichas lupus (Anarhichadidae) off the coast of Norway

The gyrodactylid monogeneans Gyrodactylus anarhichatis n. sp. and G. microanchoratus n. sp. are reported from the skin and fins of wild Atlantic wolf-fish Anarhichas lupus L. caught off the coast of northern Norway. G. anarhichatis n. sp. is also reported from farmed Atlantic wolf-fish and spotted wolf-fish A. minor Olafsen. G. anarhichatis n. sp. most closely resembles G. corti Mizelle & Kritsky, 1967, but is also similar to G. perlucidus Bychowsky & Poljansky, 1953 and G. errabundus Malmberg, 1970, while G. microanchoratus n. sp. differs from most other Gyrodactylus spp. by having marginal hooks longer than anchors and a peculiar lump on the marginal hook sickle toe pointing towards the sickle point.     

Argentophilic structures as a diagnostic criterion for the discrimination of species of the genus Gyrodactylus von Nordmann (Monogenea)

The mapping of argentophilic surface sensory structures (chaetotaxy) is a technique which has contributed to the taxonomy of larval digeneans. Four species of Gyrodactylus from four different hosts (Salmo salar, Gasterosteus aculeatus, Poecilia reticulata and Carassius auratus) were investigated using this technique in order to ascertain its usefulness in distinguishing species of this monogenean genus. A formula for the arrangement of the natural aggregations of sensilla similar to those prepared for cercariae is proposed. Although the greatest concentration of sensilla occur adjacent to the anterior extremity of the body, it is the region just posterior to the pharynx which most readily permits the discrimination of Gyrodactylus spp. based on the number and spatial configuration of these sensilla. Maps have been constructed to demonstrate these differences. It is concluded that chaetotaxy shows potential as a simple and reliable method of distinguishing species of this genus.     

First report of Gyrodactylus spp. (Platyhelminthes: Monogenea) in the western Mediterranean Sea: molecular and morphological descriptions

The Gyrodactylus spp. fauna on species of gobies, Pomatoschistus, Gobiusculus, and Knipowitschia (Gobiidae: Teleostei), from the western Mediterranean and Adriatic seas is strikingly similar to that found in the Baltic Sea and eastern Atlantic Ocean, both in morphology and in internal transcribed spacer (ITS) rRNA. The fauna consisted of Gyrodactylus branchialis, G. ostendicus, G. gondae, G. rugiensis, G. rugiensoides, and G. arcuatus. No new species have been found. A morphometric comparison between G. branchialis from the Mediterranean and Adriatic seas and its type locality in Ostend (Belgium) showed significant differences in ventral bar and marginal hook features. The morphometric variation was lower in G. rugiensis, whereas no significant differences were found in G. ostendicus. Gyrodactylus branchialis and G. ostendicus collected on P. microps were slightly different in the ITS rDNA (-0.6%) compared with specimens on the closely related P. marmoratus, probably reflecting ongoing speciation. A hybrid zone was identified in the Vaccarès lagoon complex (France) where both host species are sympatric. There was no clear geographic or host-related pattern in the variation found in the ITS2 rDNA in G. arcuatus sampled from P. microps, G. flavescens, Pungitius pungitius, K. panizzae, and its original host Gasterosteus aculeatus (2 polymorphic sites). Of all studied species, only G. arcuatus, G. rugiensis, and G. rugiensoides showed minor intraspecific variation in the ITS rDNA. Hence, the physical separation by a shoreline of more than 10,000 km is hardly reflected in the parasite ITS rDNA.     

Infections of Gyrodactylus bullatarudis and Gyrodactylus turnbulli on guppies (Poecilia reticulata) in Trinidad.

Gyrodactylus bullatarudis Turnbull, 1956, and Gyrodactylus turnbulli Harris, 1986, are recorded from guppies (Poecilia reticulata) from the northern mountains of Trinidad. Mixed infections of the 2 species were found at 9 localities. Gyrodactylus turnbulli had a predominantly posterior distribution on the fishes, whereas G. bullatarudis was more anteriorly distributed. This is the first record of these species from guppies collected from within their original range.     

Infestations by Gyrodactylus sp. of Atlantic salmon, Salmo salar L., in Norwegian rivers

Young Atlantic salmon, Salmo salar , brown trout, Salmo trutta , and Arctic char, Salvelinus alpinus , from eight rivers in North and Mid Norway were examined for Gyrodactylus. The fish were collected from 1975 to 1980. A Gyrodactylus salaris type was observed infecting salmon from six of these rivers. No trout or char were infected. A high frequency and intensity of infection of salmon were observed in all but one of the rivers surveyed. In the Saltdalselva, only one specimen of Gyrodactylus infecting one fish was observed. No subsequent mortality of salmon was observed in this river while there were signs of a high mortality of salmon in the other rivers. The salmon parr were more frequently attacked than the fry, the mortality of salmon seemed to have happened in most of the rivers one year after the first observations of Gyrodactylus were made. The mortality of salmon and the high infection rate of Gyrodactylus in these rivers appears unique, and as far as known to the authors there are no other described cases of mortality due to Gyodactylus in Atlantic salmon in natural waters. The reasons for the outbreak of Gyrodactylus in these rivers are not known. Two theories are discussed: one that the fish were weakened by environmental factors and the other that Gyrodactylus was introduced from some of the infected salmon hatcheries in Scandinavia.