荆州碘泡虫(粘体动物门,碘泡虫科)重描述及其基于18S rDNA系统发育关系分析

本实验利用现行主流分类衍征,重新对荆州碘泡虫(Myxobolus kinchowensis)进行了详细描述,其分类特征如下:包囊圆形,寄生于鲫肌肉与肾脏,肌肉包囊大小(126.7±1.8)μm,肾脏包囊直径为94.2μm;两寄生部位各形态参数无显著差异,成熟孢子正面观呈梨形,缝面观呈纺锤形,含一大一小两梨形极囊;无明显囊间突起,孢子后端无褶皱,无粘液膜。组织病理显示在两寄生部位均未引起严重的炎性反应,且感染强度不高,推测该种对宿主无显著影响。基于18S r DNA进化分析发现荆州碘泡虫与寄生在肌肉部位的碘泡虫属种类聚为一个大枝,然后该大枝又根据地理位置远近分为北美枝、欧洲枝以及亚洲枝。通过形态比较研究以及分子分析可确定荆州碘泡虫为有效种,其寄主为鲫,寄生部位为肌纤维间和肾小囊。此外,两组织差异明显的寄生部位出现说明荆州碘泡虫相应的放射孢子虫在宿主鲫体内存在不同的发育与移行途径,而肌肉可能为其常规寄生部位,肾脏为其异常寄生部位,这种寄生部位的转移与宿主转变可能是粘孢子虫物种多样性形成的机制。     

Infection of Myxobolus turpisrotundus sp. n. in allogynogenetic gibel carp,Carassius auratus gibelio (Bloch), with revision of Myxobolus rotundus (s. l.) Nemeczek reported from C. auratus auratus (L.)

Infection of a Myxobolus species, previously identified as Myxobolus rotundus, was detected in 182 of 7892 (2.31%) allogynogenetic gibel carp, Carassius auratus gibelio, in a closed pond culture system in China. Morphological and molecular data showed that this myxosporean is a different species from M. rotundus parasitizing Abramis brama in Europe and is thus designated as a new species, Myxobolus turpisrotundus. M. rotundus (s.l.) ex C. auratusauratus is a synonym of M. turpisrotundus. Plasmodia of M. turpisrotundus develop in the subepidermal tissues of the body surface resulting in an unaesthetic appearance and causing severe economic losses. Prevalence of infection with the myxosporean plasmodia varied seasonally, increasing in winter and decreasing in spring. Prevalence was positively correlated to host size, but no host sex‐specificity was found. No infection was observed in other fish species (grass carp, bighead carp and yellow catfish) reared in the same pond, suggesting that the parasite has a relatively strict host specificity. Plasmodia grew gradually as the parasite developed, and reached up to a maximum 5.6 mm in diameter. Plasmodia ruptured naturally to release the mature spores and host fish completely recovered with no mortality. Release of spores and regeneration of lesions were not correlated with water temperature. Histology showed that plasmodia developed sub‐epidermally, and that the wall of the plasmodia was composed of a multiple complex structure, including layers of fibroblasts, a collagenous membrane, melanophores and a layer of cup‐like cells of unknown derivation and function. The cup‐like cells are in direct contact with pre‐sporogonic stages located in the peripheral parts of the large plasmodia. No severe host inflammatory response was seen.     

Morphological and molecular biological studies on intramuscular Myxobolus spp. of cyprinid fish

The validity of Myxobolus species infecting the skeletal muscles of six cyprinid fish species was studied by morphological and molecular biological methods. Intracellularly developing Myxobolus spores identified as M. cyprini from the common carp, M. musculi from the barbel, and M. pseudodispar from the roach, rudd, common bream and white bream were very similar in their shape and size. Nonetheless, in species identified as M. pseudodispar, the occurrence of spores with an asymmetrical shape was higher than in M. cyprini, while asymmetrical spores were only occasionally found in M. musculi. The DNA sequence analysis of the polymerase chain reaction (PCR)‐amplified 18S rRNA gene of Myxobolus spores from these fish showed a similar phylogeny to that of their host species. As morphological studies and DNA sequence analysis demonstrated slight but real differences in the spores infecting muscles of the six cyprinid species, it is suggested that M. musculi, M. pseudodispar and M. cyprini are valid species.     

The susceptibility of diverse species of cultured oligochaetes to the fish parasite Myxobolus pseudodispar Gorbunova (Myxozoa)

This study provides detailed information on the invertebrate hosts of Myxobolus pseudodispar (Myxozoa) and explores the susceptibility range of several species and analyses the relevance of the species composition of an oligochaete population. Our findings demonstrate that the oligochaete host range of M. pseudodispar is similarly wide as the number of vertebrate host species. Besides Tubifex tubifex and Limnodrilus hoffmeisteri, Psammoryctides barbatus and Psammoryctides moravicus were also found to be susceptible invertebrate hosts. The genetic characterization of the mitochondrial 16S rDNA of T. tubifex sensu lato revealed that lineages I, II and III are susceptible to M. pseudodispar, whereas T. tubifex lineage VI seems to be non‐susceptible. T. tubifex lineage V and L. hoffmeisteri specimens were positive in a M. pseudodispar‐specific PCR, but in most cases, the release of mature actinospores could not be detected. Hence, these non‐susceptible oligochaetes likely serve as `biological filters` as they remove myxospores from the sediment without producing actinospores. Together with the phylogenetic analysis of the susceptible and non‐susceptible oligochaete hosts on the basis of mt 16S rDNA sequences, the route of the development of M. pseudodispar in the oligochaete hosts was tracked by in situ hybridization. According to our findings, the gut epithelia seem to be a portal of entry of the sporoplasms, where the development of the parasite also takes place. The basal lamina seems to be involved in the migration of the parasite, and the worm’s cellular immune response is activated by the infection.     

引起池养异育银鲫高死亡率的粘孢子虫新种咽碘泡虫形态和分子分析

对近几年发生在江苏省盐城地区引起池养异育银鲫大批死亡的粘孢子虫进行了形态和分子等方面分析研究,发现该粘孢子虫主要特点是:孢子梨形,部分孢子后部外周包围有透明无色的膜状鞘和壳瓣底部内侧呈现1～6个"V"形缺刻,两个极囊大小不等,孢子前端内侧两个极囊间有一个细长的囊间突起,胚质中无嗜碘泡,孢囊大小10～25 mm,孢子长16.82±0.43(16.03～17.69)μm,孢子宽10.26±0.43(9.12～10.88)μm,孢子厚8.09±0.29(7.50～8.75)μm,孢子长宽比1.64±0.09(1.50～1.93);大极囊长8.66±0.25(8.25～9.16)μm、宽3.65±0.25(3.01～4.19)μm,小极囊长8.35±0.28(7.60～8.85)μm、宽3.58±0.23(3.09～4.11)μm,囊间突起长1.80±0.12(1.59～2.00)μm。特异性地寄生在异育银鲫的口咽腔上颚的咽组织内、具有寄主和寄生在咽部的专一性,PCR扩增得到1 576 bp的18S rDNA序列,GenBank登录号为JQ726700。该粘孢子虫与相近的其它粘孢子虫比较,孢子长分别与白鲟碘泡虫(Myxobolus psephurusi)和扭曲碘泡虫(M.twistus)无显著差异(P>0.05),与M.ampullicapsulatus、M.wulii和鳙碘泡虫(M.aristichthydis)分别有显著差异(P<0.05);孢子宽和厚、极囊长和宽分别与M.ampullicapsulatus、M.wulii、白鲟碘孢虫(M.psephurusi)、鳙碘泡虫(M.twistus)和扭曲碘孢虫(M.aristichthydis)有显著差异(P<0.05);分子系统树分析结果显示,与M.ampullicapsulatus在同一个分支中,亲缘关系最近。从孢子形态学、寄主的特异性、寄生部位的专一性以及18SrDNA序列等特点综合比较分析,表明该粘孢子虫为一新种,命名为咽碘泡虫Myxobolus pharynae n.sp.。     

Description and phylogeny of Ceratomyxa anko sp. n. and Zschokkella lophii sp. n. from the Japanese anglerfish, Lophius litulon (Jordan)

Two new species of myxozoans from the Japanese anglerfish, Lophius litulon, are described using myxospore morphology and small subunit rDNA sequences. Ceratomyxa anko sp. n. is a parasite of the gall bladder and had a prevalence of 57%. Mature spores of C. anko sp. n. are arcuate to crescent shaped with valves tapering to rounded tips. A prominent sutural line runs centrally between the round adjacent polar capsules containing the polar filament coiled two to three times. Spore measurements: length 10.8 (9.7-11.9) microm, width 41.9 (36.9-47.2) microm, polar capsule diameter 4.6 (4.1-5.3) microm. Ceratomyxa anko sp. n. can be distinguished from other Ceratomyxa spp. due to its spore dimensions and shape. Zschokkella lophii sp. n. is a parasite of the urinary bladder and had a prevalence of 70%. Mature spores are ellipsoidal to semicircular with bluntly pointed ends. The sutural line is curved or sinuous and the valves have no discernable surface ornamentation. Two almost spherical polar capsules are located separately in the ends of the spore, opening in almost opposite directions and contain the polar filament with five coils. Spore measurements: length 20.1 (16.8-24.0) microm, width 14.9 (12.7-16.8) microm, polar capsule diameter 5.1 (3.6-5.8) microm. Zschokkella lophii sp. n. can be distinguished from other Zschokkella spp. due to the terminal opening of the polar capsules within the spores and the site of infection within the host fish. In the phylogenetic analyses, C. anko sp. n. grouped with other members of the same genus forming a monophyly. Zschokkella lophii sp. n. forms a discrete clade with another Zschokkella sp. that infects the urinary bladder of marine fish. This grouping forms a sister clade to one containing members of the genus Parvicapsula, all of which are parasites of the urinary system in marine fish.     

Alpine and Apennine barriers determining the differentiation of the rudd (Scardinius erythrophthalmus L.) in the Italian peninsula

Abstract –  A phylogeographic study of rudd Scardinius erythrophthalmus L. (Pisces: cyprinidae) was addressed to determine the differentiation of Italian populations in relation to the presence of mountain barriers, which identify two biogeographic districts (Padano-Venetian, Tuscano-Latium). At this scope, 409 bp long sequences of mtDNa cytochrome b gene were obtained from Italian and central European samples. Italian rudds phylogeny showed low level of divergence which although resulted from the central European haplotypes. The genetic structure of haplotypes in the studied districts revealed a significant recent fragmentation event. The Padano-Venetian populations showed patterns of past range expansion caused by the specie dispersion by river connections occurred during Pleistocene. This pattern provides evidence for (i) the role of Alps as a barrier for rudd dispersion; (ii) a significant genetic structure among the studied districts related to recent isolation events by the Apennine barrier; (iii) the role of Pleistocene sea level variations in determining phylogeography of the Padano-Venetian populations.     

<Emphasis Type="Italic">Myxobolus nagaraensis</Emphasis> n. sp. (Myxozoa: Myxosporea) causes abdominal distension of freshwater goby <Emphasis Type="Italic">Rhinogobius</Emphasis> sp. OR type from the Nagara River

ABSTRACT:   A new myxosporean parasite was found in the body cavity and caudal peduncle of the freshwater goby Rhinogobius sp. Orange type (OR) collected from the Nagara River, Gifu Prefecture, Japan. Infected fish exhibited substantial swelling of the abdomen caused by large parasitic cysts approximately 10 mm in size, formed in the visceral cavity. The cyst was a compacted aggregate of several smaller cysts, similar to a bunch of grapes in appearance. Histological examination showed that plasmodia developed within the renal capsule, and finally occupied the visceral cavity. Spores were ovoid with an attenuated anterior end. Sutural ridges were conspicuous with several folds on the edge. Average spore size was 11.9 (10.5–13.5) μm long, 9.0 (8.0–10.0) μm wide, and 6.5 (6.0–7.0) μm thick. Two equal polar capsules were 5.5 (4.5–6 0) μm long and 3.0 (2.5–4.0) μm wide. Partial small subunit ribosomal DNA sequences of the myxosporean were distinct from those of other myxozoan species in GENBANK. A new species name, Myxobolus nagaraensis , is proposed for this parasite.     

The life cycle of Chloromyxum auratum (Myxozoa) from goldfish, Carassius auratus (L.), involves an antonactinomyxon actinospore

The myxozoan parasite Chloromyxum auratum Hallett, Atkinson, Holt, Banner & Bartholomew, 2006 , was shown experimentally to have a two‐host life cycle which involved a previously undescribed antonactinomyxon actinospore stage. Myxospores obtained from gall bladders of naturally infected feral goldfish, Carassius auratus (L.), were used to infect samples of mixed species of oligochaete worms obtained from the same locality as the fish: Fern Ridge Dam, Oregon, USA. After some 110 days post‐exposure, actinospores were detected from the water above the oligochaetes. The 18S rDNA sequence of these actinospores was identical to the original myxospores. Spore release was sporadic, of low intensity and short duration, which confounded efforts to identify the host oligochaete species and infect naïve fish. This is the first life cycle that incorporates an actinospore of the collective group Antonactinomyxon, and the first life cycle demonstrated in the laboratory for a species of Chloromyxum.     

Habitat use and activity patterns of roach (Rutilus rutilus (L.)), rudd (Scardinius erythrophthalmus (L.)), perch (Perca fluviatilis L.) and pike (Esox lucius L.) in the laboratory: the role of predation threat and structural complexity

Abstract– The role of predation risk and structural complexity in determining the habitat use and activity patterns of roach, rudd and perch was assessed using a series of video-recorded laboratory trials. The time spent in open and structured habitats, vertical swimming heights and activity levels of each species were observed in the presence and absence of a potential, pike, predator. Habitat complexity varied between treatments with artificial stem densities of 200, 400 and 600 stems'm⁻². Predator free trials showed that roach and rudd spent significantly less time in structured habitats than perch. Increasing stem density had no significant effect on the habitat choice of perch but did affect the distribution of roach and rudd. Stem density influenced the vertical swimming height of rudd or perch but not roach, although the effects of habitat complexity on swimming activity were more complicated. Pike were themselves influenced by increases in stem density, only selecting structured habitats when stem densities were less than 600 stems'm⁻². In the presence of a predator, both roach and rudd increased the amount of time spent in structured areas. These observed differences were independent of stem density. Perch, however, decreased the amount of time spent in structural habitats at all stem densities. Predation risk also prompted significant changes in both vertical swimming height and activity levels of potential prey fish. A pike predation success hierarchy of rudd-roach-perch was also observed during the study. The reasons for this pattern are discussed.     

Morphology,seasonality and phylogeny of Zschokkella soleae sp. n. (Myxozoa,Myxosporea) parasite of Solea solea (L.) (Pleuronectiformes,Soleidae) from Ghar El Melh Lagoon,Tunisia

A new Myxosporea, Zschokkella soleae sp. n., was found in the gall bladder and the bile of common sole, Solea solea (L.), from Ghar El Melh Lagoon in north‐east Tunisia. This is the first record for the presence of Zschokkella species in Tunisian waters. The parasite's plasmodia are polysporic with variable size and shape. Some plasmodia appeared attached to the gall bladder epithelium while others were found free in bile. Mature spores are ellipsoidal in frontal view 13.8 ± 0.38 μm long and 10.86 ± 0.40 μm wide with two equal size spherical polar capsules 3.6 ± 0.43 μm in size. The prevalence of infection seems to correlate with host size and changes over the year with maximum percentage in summer. Based on the 18S rDNA sequence data, Z. soleae sp. n. is readily distinguishable from other myxozoan DNA sequences in GenBank. Phylogenetically, the new species is placed in the freshwater Myxidium clade including several Zschokkella spp. infecting the gall bladder. Morphology, histology as well as DNA sequence analysis indicate that the examined species differs from all previously described Zschokkella species.     

An updated geographic distribution of Myxobolus cerebralis (Hofer, 1903) (Bivalvulida: Myxobolidae) and the first diagnosed case of whirling disease in wild-caught trout in the south-eastern United States

Myxobolus cerebralis (Bivalvulida: Myxobolidae), the aetiological agent of salmonid whirling disease, was detected in 2 river basins of North Carolina during 2015, which initiated the largest spatial–temporal monitoring project for the disease ever conducted within the south-eastern United States (focused mainly in eastern Tennessee and western North Carolina). A total of 2072 rainbow trout Oncorhynchus mykiss, 1,004 brown trout Salmo trutta and 468 brook trout Salvelinus fontinalis were screened from 113 localities within 7 river basins during June 2017 through October 2019. Infections were detected by pepsin–trypsin digest, microscopy and the species-specific nested polymerase chain reaction (PCR) in 19 localities across 6 river basins. Myxospore morphology was indistinguishable from the published literature. In 2019, five rainbow trout that symptomatic for whirling disease (sloping neurocranium and lordosis) were captured and processed for histopathology. Myxospores were detected in the calvarial cartilage of two deformed trout with associated erosion of the cartilage consistent with reported whirling disease lesions. This is the first report of M. cerebralis in Tennessee and the first histologically confirmed cases of whirling disease in southern Appalachian (south-eastern United States) rivers and streams and expands the distribution of M. cerebralis throughout western North Carolina and eastern Tennessee.     

Henneguya pseudorhinogobii n. sp. (Myxozoa: Myxosporea) parasitizing the gills of the freshwater goby Rhinogobius sp. OR from the Nagara River and redescription of Henneguyarhinogobii

Two types of infection with myxosporean parasites, which were different in cyst size, were found from the gill of Rhinogobius sp. OR collected from the Nagara River, Gifu Prefecture, Japan. A myxosporean forming large-type cysts in the gill filament was morphologically identified as Henneguya rhinogobii. Another myxosporean forming small-type cysts was found to be parasitic in the gill lamella. Average spore sizes of the latter myxosporean were 15.8 (range 14.2–17.8) μm in length, 5.3 (4.7–5.8) μm in width, 6.5 (5.9–7.6) μm in polar capsule length and 34.9 (25.3–42.9) μm in caudal appendage length, which were almost identical to those of H. rhinogobii. However, the two were distinguishable by the location in the gill (intrafilamental or intralamellar) and by the difference in the sequence of SSU rDNA (about 96% similarity). We propose the myxosporean forming intralamellar small-type cysts as Henneguya pseudorhinogobii n. sp. and redescribe H. rhinogobii by morphological and molecular studies.     

Description of Myxobolus bulbocordis sp. nov. (Myxosporea: Myxobolidae) from the heart of Barbus sharpeyi(Günther) and histopathological changes produced by the parasite

A new myxosporean, Myxobolus bulbocordis sp. nov., has been found in the heart of a Mesopotamian fish, Barbus sharpeyi (Günther), in rivers in Southwest Iran. Cysts containing matured spores were located on the serosa of the atrium cordis, bulbus arteriosus, larger gill arteries and inside the wall of the bulbus. The species showed an affinity to connective tissue cells and was never associated with the muscles. Mature cysts were surrounded by a connective tissue capsule composed of two to three layers. Spores in disrupted cysts were infiltrated by epithelioid cells and macrophages. The spores found in Barbus sharpeyi differed in size and morphology from species known from other barbels.     

Gastric coccidiosis in Thoracocharax stellatus caused by Goussia guamaensis n. sp. (Apicomplexa: Eimeriidae) from the Amazon region

Ninety‐seven specimens of spotfin hatchetfish, Thoracocharax stellatus, an ornamental freshwater species from the Amazon basin, were captured in the basin of the Guamá River in the municipality of Belém, in northern Brazil, and analysed for coccidiosis infection. Overall, 26 of the specimens were infected by apicomplexan parasites of the genus Goussia, with unsporulated forms being found in the gastric epithelium and sporulated forms in the intestinal lumen. The spheroid oocysts (mean diameter: 13.2 ± 1.7 μm) have four elliptical sporocysts. A partial sequence of the SSU rDNA of the new species was obtained, which contained 1,121 base pairs, with 43.8% guanine + cytosine (G + C), and the bases distributed as follows: A = 28.1%, C = 18.3%, G = 25.5% and T = 28.1%. The combined analysis of the morphometric and phylogenetic evidence confirmed that the specimens represented the genus Goussia and were allocated to a new species, Goussia guamaensis n. sp., which is described here.     

Unicauda magna sp.n. (Protozoa: Myxozoa): a new myxozoan from the fin tissues of the fathead minnow, Pimephales promelas Rafinesque

Abstract. Unicauda magna sp.n. (Protozoa: Myxozoa) was found in hatchery-raised fathead minnows, Pimephales promelas (Cyprinidae) from Pennsylvania. A large number of whitish-coloured cysts were located in the tissues of the pectoral, pelvic and caudal fins of infected fish. Spore dimensions (in μm) based on 30 measurements (10 spores/cyst) included the following: total length, 109.6 (75–170); spore body length, 15.5 (14–17); spore body thickness, 12.1 (10–13); spore body width, 13.2 (12–14.5); tail length, 94.1 (60–154); polar capsule lengths, 8.6 (7.5–9.5); polar capsule width 4.1 (3.0–5.0); sutural ridge width, about 1. The tail and body of the spores differed in colour when stained with each of three dyes. They are, therefore, composed of different material.     

珠带鱼和带鱼未定种Trichiurus sp.2的分子系统进化关系

针对南海大型带鱼新种——珠带鱼(Trichiurus margarites Li,1992)与日本产带鱼未定种T.sp.2(sensu Nakabo,2002)之间的系统进化关系及分类地位不明确问题,本研究通过测定采自海南岛西岸珠带鱼的线粒体16S rRNA基因部分序列,结合已报道的T.sp.2及其近缘种的同源序列,对其16S rRNA基因序列变异进行分析。基于Kimura双参数模型计算,海南岛西岸珠带鱼与日本九州、海南岛东岸及印度洋T.sp.2间的净遗传距离(0.000～0.006)远低于属内其他种间的净遗传距离(0.037～0.061),表明二者的遗传分化程度处于种内水平。邻接法(NJ)和最大似然法(ML)构建的分子系统树显示,珠带鱼与T.sp.2以极高的置信度(NJ 100%,ML 96%)聚为一支单系,并与其他带鱼形成姐妹群。根据所得分子数据并结合形态学研究资料,可以确定日本产带鱼未定种即是珠带鱼,它广泛分布于印度西太平洋。综合3种带鱼属鱼类在西北太平洋的种间分化事件,对珠带鱼的起源及演化进行了初步推测。