Specific and common antigens of Clonorchis sinensis and Opisthorchis viverrini(Opisthorchidae,Trematoda)

The antigenic characterizations and serological reactions of human liver flukes, Clonorchis sinensis and Opisthorchis viverrini, were analyzed by immunoblot. The antigenic profiles of the crude extract of Clonorchis contained major proteins of 8, 26-28, 34-37, 43, and 70 kDa, and those of Opisthorchis 34-37, 43, 70, and 100 kDa. Of these, the 8, 26-28 and 34-37 kDa bands of Clonorchis and the 100 kDa of Opisthorchis were major components of each excretory-secretory antigen. The 8 and 26-28 kDa bands were specific to Clonorchis but the 100 kDa of Opisthorchis cross-reacted with the sera of clonorchiasis, and the 34-37, 70 and 100 kDa bands cross-reacted with sera of other helminthiases. The frequency and intensity of the immunoblot reactions were positively correlated with the intensity of the liver fluke infection.     

The ultrastructural characterization of the tegument of Clonorchis sinensis (Cobbold, 1875) cercaria.

The tegumental ultrastructure of the cercaria of the liver fluke, Clonorchis sinensis (Cobbold, 1875), was studied by scanning and transmission electron microscopy. The body surface is almost encircled by many rows of regularly arranged spines. The tegumental syncytium of the body contains many rod-shaped dense granules and central electron-lucent bodies, neither of which are present in the tail tegument. There are four rows of hooked teeth and modified spines on the oral cone. These teeth are differentiated morphologically and probably functionally from the other body spines. Disc-shaped papillae with long or short cilia are distributed on the body in a bilaterally symmetrical pattern dorsally and ventrally. There are 30 to 37 papillae with much longer cilia laterally. Four pairs of papillae were found on the tail. From their structure and location these papillae appear to have a mainly tango- or rheoreceptive function. Another type of sheathed papillae is situated around the oral sucker. The cuticular tegument is expanded laterally at the base of the tail, forming a characteristic sac-like structure.     

The ITS1-5.8S-ITS2 sequence region in the Musaceae: structure, diversity and use in molecular phylogeny

Genes coding for 45S ribosomal RNA are organized in tandem arrays of up to several thousand copies and contain 18S, 5.8S and 26S rRNA units separated by internal transcribed spacers ITS1 and ITS2. While the rRNA units are evolutionary conserved, ITS show high level of interspecific divergence and have been used frequently in genetic diversity and phylogenetic studies. In this work we report on the structure and diversity of the ITS region in 87 representatives of the family Musaceae. We provide the first detailed information on ITS sequence diversity in the genus Musa and describe the presence of more than one type of ITS sequence within individual species. Both Sanger sequencing of amplified ITS regions and whole genome 454 sequencing lead to similar phylogenetic inferences. We show that it is necessary to identify putative pseudogenic ITS sequences, which may have negative effect on phylogenetic reconstruction at lower taxonomic levels. Phylogenetic reconstruction based on ITS sequence showed that the genus Musa is divided into two distinct clades--Callimusa and Australimusa and Eumusa and Rhodochlamys. Most of the intraspecific banana hybrids analyzed contain conserved parental ITS sequences, indicating incomplete concerted evolution of rDNA loci. Independent evolution of parental rDNA in hybrids enables determination of genomic constitution of hybrids using ITS. The observation of only one type of ITS sequence in some of the presumed interspecific hybrid clones warrants further study to confirm their hybrid origin and to unravel processes leading to evolution of their genomes.     

The placement of South African strains of Beauveria in a phylogeny inferred from rDNA ITS1-5.8S-ITS2 sequences

This study aimed to confirm the identity of three strains of the entomopathogenic fungus Beauveria bassiana from South African soils and to investigate their phylogenetic relationship with non-indigenous strains from other geographic regions. Sequences of the rDNA ITS1-5.8S-ITS2 region of 23 strains were compared with the Genbank reference sequences of 20 other cosmopolitan strains. Fitch parsimony and neighbor-joining analyses of the ITS1-5.8S-ITS2 regions resolved the strains into two distinct clades and matched them to four species groups/lineages: Beauveria bassiana, B. cf. bassiana (pseudobassiana), B. brongniartii and B. caledonica. Two of the South African strains initially identified as B. bassiana grouped with B. caledonica, whereas the third strain was confirmed as B. bassiana. Because of the paucity of Genbank references for B. caledonica, we have designated the two South African B. caledonica strains as B. sp. aff. caledonica. Other reassignments included two strains from Norway, originally classified as B. bassiana, being grouped with B. brongniartii, and three of the B. brongniartii reference taxa from Brazil which were clearly placed in the B. bassiana clade. The study provides a first report of the presence of the B. caledonica lineage in Africa and confirms current Beauveria phylogenies inferred from molecular data.     

Relationships amongOmphalotus species (Paxillaceae) based on restriction sites in the ribosomal ITS1-5.8S-ITS2 region

Genetic relationships for several species of the fungusOmphalotus were estimated by comparing the presence or absence of restriction sites in the ITS1-5.8S-ITS2 region of nuclear ribosomal DNA. Results placeO. olearius, O. subilludens andO. olivascens in a single clade.Omphalotus illudens, usually thought to be related toO. subilludens, was placed in a second clade, more closely related toO. nidiformis. Omphalotus mexicana is distinct from all other examinedOmphalotus species.     

绒泡菌目黏菌的ITS1-5.8S-ITS2二级结构的比较分析

为探讨核糖体DNA转录间隔区(r DNA ITS)的RNA二级结构在黏菌系统发育研究中的作用,以黏菌ITS通用引物PHYS4和PHYS5对绒泡菌目5属8种黏菌的r DNA ITS进行扩增和测序,利用RNA structure构建了ITS区的RNA二级结构模型。结果表明:ITS1在绒泡菌目黏菌中不能形成一个紧实的结构,但大部分物种都具有一段稳定的螺旋结构,可能对r RNA的成熟具有作用;5.8S r RNA的二级结构相似,由4个螺旋组成,主要为两种类型;基于5.8S r RNA和28S r RNA相互作用构建的ITS2的二级结构模型显示,它由一个封闭的多分支环和至少4个主要的螺旋组成,其中螺旋IV结构相对比较保守。由于ITS区的二级结构相比核苷酸序列更加保守,因此深入地分析其二级结构有助于认识其结构与进化的关系。     

Genetic and morphologic diversity of the moles (Talpomorpha,Talpidae, Mogera) from the continental Far East

Taxonomy of the East Asian moles of the genus Mogera is still controversial. Based on the sequence data of 12 nuclear genes and one mitochondrial gene, we examine genetic variation in the Mogera wogura species complex and demonstrate that M. robusta, from the continental Far East, and M. wogura, from the Japanese Islands, are not conspecific. Our data do not support the existence of two or more species of Mogera in the Russian Far East. We suggest that the form “coreana” from the Korean Peninsula should be treated as a subspecies of M. robusta. Our morphological analysis shows that M. r. coreana differs from typical M. robusta, from Primorye, primarily in its smaller size. We show that there is strong morphological variability among continental moles, which may be associated with ecological and geographical factors. The time since the split between M. wogura s. str. and M. robusta dates back to the Middle Pleistocene (0.30–1.0 Myr), while M. r. coreana separated from M. r. robusta in the Upper Pleistocene (0.04–0.18 Myr). The results of the mismatch analysis indicate recent demographic expansion in populations of moles from Primorye.     

Evolutionary insights inferred by molecular analysis of the ITS1-5.8S-ITS2 and IGS Avena sp. sequences

In an attempt to clarify phylogenetic and genome relationships among 35 diploid (A and C genomes), 13 tetraploid (AB and AC genomes) and 6 hexaploid (ACD genome) Avena taxa, 71 clones of the ITS1-5.8S-ITS2 fragment were sequenced, aligned and a network was constructed. In addition, the intergenic spacer (IGS) fragment was fingerprinted by means of a RFLP analysis using three different restriction enzymes. Both approaches led to comparable results. Clustering among the 54 Avena sp. entries was according to karyotype. Major genic divergence between the A and C genomes was revealed, while distinction among the A and B/D genomes was not possible. High affinity among the AB genome tetraploids and the A(s) genome diploid A. lusitanica was found, while AC genome tetraploids and ACD hexaploids were highly affiliated with the A(l) genome diploid A. longiglumis. The possible role of A. longiglumis in Avena sp. evolution is discussed.     

Molecular phylogeny ofSalicaceae and closely relatedFlacourtiaceae: Evidence from 5.8 S, ITS 1 and ITS 2 of the rDNA

A ribosomal DNA region, including the entire 5.8S RNA gene and the internal transcribed spacers ITS 1 and ITS 2, was used for studying the phylogeny ofSalicaceae and the relationship betweenSalicaceae andFlacourtiaceae. The length of the ITS regions withinSalicaceae andFlacourtiaceae was similar to that found in other angiosperms. The GC content of both ITS regions was high, varying 62.7-72.2%. The most parsimonious tree clusters the wind-pollinatedChosenia bracteosa among theSalix species, suggesting that it should be included in the genusSalix. The grouping withinSalix leaves subg.Salix as paraphyletic, for which reason the subgeneric division is questionable.Populus was monophyletic and formed a sister group toSalix. The interspecific variation of the ITS sequences was very small inSalicaceae, which is in contradiction to the age of the group according to the evidence from fossil data.Idesia polycarpa fromFlacourtiaceae shows great sequence similarity withSalicaceae, but the analysis of 5.8S rDNA supports monophyly of the four species ofFlacourtiaceae sampled for this study.     

The life cycle of Asymphylodora perccotti sp. n. (Trematoda: Lissorchiidae) in the Russian Southern Far East

Specimens of Asymphylodora perccotti sp. n. (Trematoda: Lissorchidae) were found in the esophagus of the freshwater fish Perccottus glenii (Odobantidae) taken from the Bolshaya Ussurka River Basin (Primorsky Region, Russian Southern Far East). The first intermediate host of this trematode is a gastropod, Parafossarulus manchouricus, and the secondary hosts are the same mollusk and Boreoelona ussuriensis. Specimens of the new species are similar to A. amnicolae identified by Stunkard in 1959, but the mature worms have larger suckers and shorter ceca. The cercariae of these species are distinguished by body, suckers and pharynx size. These organs in A. perccotti sp. n. are more than one-third larger than what is observed in A. amnicolae. In addition, the new species lacks the capacity for progenesis. Finally, the new species is unusual in that it resides in the fish esophagus instead of the intestine, as is common for most Asymphylodora species. Partial ribosomal DNA sequences and phylogenetic reconstruction sequence data indicate that these worms represent a new digenean species.     

Proposed division of the genus Heterophyes Cobbold, 1865 (Trematoda: Heterophyidae)

A new genus, Alloheterophyes, is erected for Heterophyes chini Pearson & Pearson, 1983 and for H. superspinata Leonov & Belogurov, 1965, with which H. bitorquata Pearson & Pearson, 1983 is synonymised. New host and locality records are given. Alloheterophyes differs from Heterophyes in having: (i) a seminal vesicle of two tandem chambers, the distal with a thicker wall; (ii) a uterine seminal receptacle; (iii) an unspined genital atrium; (iv) an inflated, almost triangular excretory bladder; (v) a ventral sucker not close to the gut fork; (vi) the final loop of the uterus dextral to the gonotyl; and (vii) a clavate shape.     

Allocreadium khankaiensis sp.nov. and Allocreadium hemibarbi Roitman, 1963 (Trematoda: Allocreadiidae) from the Russian Far East: Morphological,molecular, and phylogenetic studies

Three species of the genus Allocreadium were collected from cyprinid fish in the territory of the Russian Far East: Allocreadium khankaiensis sp.nov; Allocreadium hemibarbi Roitman, 1963, and Allocreadium sp. 1. It is established that, according to morphometric values, the species A. khankaiensis sp.nov. has high morphological similarity with Allocreadium elongatum Achmerov, 1960; Allocreadium erythroculteris Achmerov, 1960; and Allocreadium aburahaya Shimazu, 2003. However, A. khankaiensis sp.nov. differs from A. elongatum and A. erythroculteris by vitellaria extension and size of cirrus pouch. In addition, A. khankaiensis sp.nov. differs from A. erythroculteris by the smaller size of the body and ventral sucker. A. khankaiensis sp.nov. and A. aburahaya have similar metrical values but A. khankaiensis sp.nov. differs by the shape of the testes: rounded vs. lobed. Species affiliation of another Allocreadium species is based on the morphometric identity with the species A. hemibarbi. Phylogenetic tree reconstruction using partial sequences of the 28S rRNA gene revealed a clear resolution of all Allocreadium species, excluding Allocreadium lobatum and Allocreadium neotenicum. The genetic analysis detected Allocreadium sp. 1 as a presumably independent species from the Razdolnaya River basin.     

基于ITS1-5.8S rRNA-ITS2序列的猪源结肠小袋纤毛虫种群特征分析

【目的】为了揭示结肠小袋纤毛虫病在环境中的分子传播机制,研究了猪源结肠小袋纤毛虫的种群特征。【方法】用粪便涂片镜检和改进型DMEM培养基从病猪结肠内容物分离结肠小袋纤毛虫滋养体,然后用显微观察、吖啶橙荧光染色法和基于ITS1-5.8S rRNA-ITS2序列的分子标记技术分析了豫西地区猪群中结肠小袋纤毛虫的种群特征。【结果】结果显示,从来自豫西地区8个县市的病猪中共分离了15株小袋纤毛虫,5.8S rRNA序列相似性高达99.4%以上,同属于结肠小袋纤毛虫,根据ITS1/2序列分析结果,MJ-2和SX-1株属于结肠小袋纤毛虫基因型A,其余13株均属于结肠小袋纤毛虫基因型B。MJ-2和SX-1株滋养体形态特征明显区别其他13株,绝大多数呈球形,运动缓慢,粪便中和体外培养的虫体密度较低;而其他13株的滋养体均呈多形性,运动快速活跃,虫体密度较大。吖啶橙荧光染色显示,2种基因型滋养体的核结构没明显差别。【结论】首次报道中国猪源结肠小袋纤毛虫存在2个基因型,其中基因型B为优势种群,为防控人和动物结肠小袋纤毛虫病提供重要参考。     

Genetic diversity of the striped field mouse (Apodemus agrarius Pallas, 1771) in the Russian Far East as assessed by RAPD-PCR

Random amplified polymorphic DNA (RAPD) analysis based on polymerase chain reaction (PCR) with ten-nucleotide primers of arbitrary sequences (RAPD-PCR) was used to study the genetic characteristics of five samples of the striped field mouse (Apodemus agrarius Pallas, 1771) from the Russian Far East (Primorye, Khabarovsk krai, and Magadan oblast). Highly significant differentiation of the samples was demonstrated, the genetic diversity of each sample was estimated, and non-neutral loci were found. The genetic diversity was the highest in a population from the outskirts of Magadan and the lowest in populations from an island on the Amur River near Khabarovsk and from the village of Talon (Magadan oblast). These two last populations were found to be genetically close to each other. These results indicate that striped field mice have been repeatedly introduced to Magadan oblast. The Talon population is likely to have originated from the left bank of the Amur; the outskirts of Magadan are inhabited with a mixed population formed by mice from Primorye and other regions.     

A phylogenetic perspective on larval spine morphology in Leucorrhinia (Odonata: Libellulidae) based on ITS1, 5.8S, and ITS2 rDNA sequences

Leucorrhinia (Odonata, Anisoptera, Libellulidae) consists of 14-15 species with a holarctic distribution. We have combined the morphological characters of a previous study with sequence data from the ITS1, 5.8S rDNA, and ITS2 regions of the nuclear ribosomal repeat. Cloning was used to investigate the intra-individual variation and such variation was found in all investigated species. Parsimony jackknifing was used to identify supported groups. The effect of sequence alignment and gap coding was explored by a modified sensitivity analysis. Loss of spines in Leucorrhinia larvae has occurred twice: once in Europe and once in North America. The role of spines as a defence against predation is discussed in a phylogenetic context.     

Intraspecific phylogeography of Carchesium polypinum (Peritrichia, Ciliophora) from China, inferred from 18S-ITS1-5.8S ribosomal DNA

Based on the variation of site 34, 46, 241, 305 and 322 in the 18S-ITS1 rDNA sequence, 19 Carchesium polypinum populations collected from eight provinces of China were separated into northern and southern population along the delineation between the Yangtze River and the Pearl River. This geographic distribution pattern of Carchesium polypinum maybe results from two factors: the vicariance resulting from the formation of the delineation between the Pearl River and the Yangtze River accompanied with the uplift of Qinghai-Xizang Plateau, and the different dispersal paths of C. polypinum affected by the climate.     

Fleas (Siphonaptera) of Carnivores (Mammalia,Carnivora) of the Russian Far East

Entomological Review - Ten flea species were collected in 2012–2017 from 6 species of carnivores in Primorsky Territory and Sakhalin Province of Russia. The most complete collections of fleas...