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At least 19 recordsLinked to original sources
PubMedDate not supplied

[Use of polymerase chain reaction for identifying helminth DNA from the species Trichinella, Fasciola, Echinococcus, Nematodirus, Taenia].

We have used the polymerase chain reaction (PCR) and random amplified polymorphic DNA (RAPD) method to identify DNA polymorphism in three classes of helminths, Cestoda, Nematoda and Trematoda. In this study, RAPD markers have been used for a description of the genetic variability between the families, genera, species and isolates of helminths. We optimized the conditions of the RAPD analysis and revealed the genetic variability of helminths belonging to different taxonomic groups. These results show that the RAPD-PCR method can be considered as a suitable technic for a phylogenetic study of helminths. The problem of an age variability and most specificity of RAPDs markers is discussed.

Animals↗
PubMed1998

Immunopathological mechanisms underlying the time-course of Trichinella spiralis cardiomyopathy in rats.

The present study shows that isolated, perfused hearts from rats orally infected with Trichinella spiralis have a reduced left ventricular developed pressure (LVDP), heart rate (HR) and coronary flow (CF). This reduction is considerably enhanced by a single bolus (100 pM) of PAF (platelet activating factor, an eosinophil activator), especially at 21 days post-infection (d.p.i.), which is the time of the maximum increase in blood and tissue eosinophilia. Helminthic DNA analysis shows that, from 21 d.p.i. onwards, the morphological and functional changes in the myocardium cannot be ascribed to the parasite's presence, whereas its antigens and the attendant immunopathological reactions might have a role in the induction of myocardial damage and dysfunction. Some perivascular inflammatory cells (eosinophils and mast cells) appear to undergo degranulation. All these data suggest a complex sequence of events, from acute myocarditis (21 d.p.i.) which may lead in time (48 d.p.i. onwards) to a dilating cardiomyopathy.

Animals↗
PubMed1998

Molecular phylogeny of the families Campulidae and Nasitrematidae (Trematoda) based on mtDNA sequence comparison.

Historically, the systematic arrangement of the genera within the family Campulidae, and its relationship with its allied family Nasitrematidae have been rather confused, particularly because only adult morphology has been available to classical taxonomic analysis. In this paper we provide a partial phylogeny of the genera of these families based on mtDNA from five campulid species: Campula oblonga, Zalophotrema atlanticum, Hadwenius tursionis, Oschmarinella rochebruni and Orthosplanchnus fraterculus; and one nasitrematid, Nasitrema globicephalae. Fasciola hepatica and Dicrocoelium dendriticum were used as outgroups. Maximum parsimony and neighbour-joining methods were applied. Both methods produced similar trees where H. tursionis appeared as the basal campulid, with a sequential divergence of Z. atlanticum, N. globicephalae, C. oblonga, O. rochebruni and O. fraterculus. Results suggest that Nasitrematidae as defined should loose its familial status and the current subfamilial division of the family Campulidae is at least partly artificial and should not be maintained.

Amino Acid Sequence↗
PubMed1995

Intracellular promiscuity in Schistosoma mansoni: nuclear transcribed DNA sequences are part of a mitochondrial minisatellite region.

It has been shown that the mtDNA of the parasitic trematode Schistosoma mansoni is hypervariable in size. We report here that this length variation is due to a large polymorphic minisatellite composed of two types of repeated sequences of 558 bp and 62 bp. Each minisatellite repeat is made up of a large 558-bp component and a variable tandem array of the small 62-bp unit. Of more fundamental interest was the finding that both the 558-bp and 62-bp components have significant homology with a gene, SM750, previously identified in the nuclear genome of S. mansoni. The small 62-bp unit is identical to the nuclear polymorphic repeat element, which is apparently spread throughout the nuclear genome and is abundant among transcripts, in addition to being present in five tandem copies in SM750. The presence, in the S. mansoni mtDNA, of fragments of genes that are present in and transcribed from the nuclear genome raises the question of the origin of these sequences. The arrangement and the variability that the mtDNA minisatellite embodies were explored as an identity test for S. mansoni based on the use of PCR for tallying the relative abundance of the several repeat numbers of the tandem arrays of the 62-bp unit within the minisatellite structure.

Animals↗
PubMed1997

Phylogenetic analysis of the Monogenea and their relationships with Digenea and Eucestoda inferred from 28S rDNA sequences.

Platyhelminth phylogeny is controversial. Phylogenetic analyses of the partial domain C1 and the full domains D1 and C2 (358 nucleotides) from the 28S ribosomal RNA gene for 21 species from the Monogenea, Digenea, Cestoda, and, as the outgroup, Tricladida reveal major departures from prevailing theory. The Digenea and not the Monogenea (Monopisthocotylea and Polyopisthocotylea) form the sister group of the cestodes; the Monopisthocotylea and Polyopisthocotylea are each monophyletic, but the Monogenea do not form a monophylum; the sister group of the Digenea + Cestoda is the Polyopisthocotylea; and Monopisthocotylea are the sister group of all other parasitic flatworms.

Animals↗
PubMed1994

A trans-spliced leader RNA sequence in plant parasitic nematodes.

A trans-spliced leader gene has been identified in the genomes of the potato cyst nematodes Globodera rostochiensis and G. pallida. The gene contains a 22-nt sequence identical to that of the leader sequence of Caenorhabditis elegans, a consensus splice donor site and a putative Sm antigen binding site. In common with other nematodes the spliced leader gene is present in tandem repeating units together with the 5S ribosomal RNA gene. Variation in the length of the intergenic spacer region has permitted the design of polymerase chain reaction primers which can be used to reveal repeat length variants diagnostic for G. rostochiensis and G. pallida and the Pa1 pathotype of G. pallida.

Animals↗
PubMed1997

Molecular delineation of Cylicocyclus nassatus and C. ashworthi (Nematoda:Strongylidae).

The nucleotide sequences of the first internal transcribed spacer (ITS-1), 5.8S gene and second internal transcribed spacer (ITS-2) of ribosomal DNA have been determined for Cylicocyclus nassatus, C. ashworthi and C. insignis. Pairwise comparisons revealed sequence differences between the taxa ranging from 3.8 to 6.2% for the ITS-2 and 2.2-2.7% for the ITS-1. For the ITS-1, the level of the sequence difference between C. ashworthi and C. nassatus (2.2%) was equivalent to that between C. nassatus and C. insignis (2.2%), indicating that C. ashworthi and C. nassatus represent separate species. Theoretical restriction maps were constructed from the sequence data, and a polymerase chain reaction-linked restriction fragment length polymorphism (PCR-linked RFLP) technique was established to unequivocally distinguish C. ashworthi from C. nassatus.

Animals↗
PubMed1998

Differences in the 5.8S rDNA sequences among ascarid nematodes.

Sequences of the 5.8S rDNA were obtained for 14 species of nematode from different superfamilies and families within the order Ascaridida. All sequences were 157 bp in length. Sequence differences among species ranged from 0 to 18 bp (0-11.5%). A phenetic analysis of the sequence data groups the 14 taxa into their respective superfamilies and families, but does not discriminate fully at the subfamily level. A phylogenetic analysis of the sequence data failed to resolve the evolutionary relationships at the superfamily level. The 5.8S gene may be useful for phylogenetic studies of the phylum Nematoda at the ordinal level.

Animals↗
PubMed1995

Genetic markers for closely-related parasitic nematodes.

Seven species of closely-related nematode parasite (Trichostrongylus axei, T. colubriformis, T. probolurus, T. retortaeformis, T. rugatus, T. vitrinus and T. tenuis) were characterized using a polymerase chain reaction-linked restriction fragment length polymorphism technique (PCR-RFLP). The rDNA region spanning the first and second internal transcribed spacers as well as the 5.8S rDNA gene (ITS+) was amplified from isolates of each of the seven species, digested separately with six restriction endonucleases (Dra I, Hinf I, Rsa I, Vsp I, Nla III and Tsp 509 I) and the fragments separated by agarose gel electrophoresis. PCR-RFLP of ITS+ produced characteristic patterns for each Trichostrongylus species examined. No variation in RFLP patterns was observed among different isolates for species where multiple isolates were examined. The present study demonstrates that the ITS+ provides genetic markers for the species identification of closely-related parasitic nematodes, and indicates the usefulness of these markers for diagnostic purposes, and epidemiologic and molecular-systematic studies on parasites and other eukaryotic organisms.

Animals↗
PubMed1995

Increased frequency of deletions in the mitochondrial genome with age of Caenorhabditis elegans.

We have developed a long-extension-PCR strategy which amplifies approximately half of the mitochondrial genome (6.3 kb) of Caenorhabditis elegans using an individual worm as target. We analyzed three strains over their life span to assess the number of detectable deletions in the mitochondrial genome. Two of these strains are wild-type for life span while the third is mutant in the age-1 gene, approximately doubling its maximum life span. At the mean life span in wild-type strains, there was a significant difference between the frequency of deletions detected in the mitochondrial genome compared with the mean number of deletions in young animals. In addition, deletions in the mitochondrial genome occur at a significantly lower rate in age-1 mutants as compared with wild type. We cloned and identified the breakpoints of two deletions and found that one of the deletions had a direct repeat of 8 bp at the breakpoint. This is the largest single study (over 900 individual animals) characterizing the frequency of deletions in the mitochondrial genome as a function of age yet carried out.

Aging↗
PubMedDate not supplied

Genetic divergence of Asiatic Bdellocephala (Turbellaria, Tricladida, Paludicola) as revealed by partial 18S rRNA gene sequence comparisons.

Polymerase chain reaction (PCR) and direct sequencing of small ribosomal RNA genes were used for analysis of genetic differences among Asiatic species of freshwater triclad genus Bdellocephala. Representatives of four species and four subspecies of this genus were used to establish homology between nucleotides in the 5'-end portion of small ribosomal RNA gene sequences. Within 552 nucleotide sites of aligned sequences compared, six variable base positions were discovered, dividing Bdellocephala into five different genotypes. Sequence data allow to distinguish two groups of these genotypes. One of them unites species from Kamchatka and Japan, another one unites Baikalian taxa. Agreement between available morphological, cytological and sequence data is discussed.

Animals↗
PubMed1997

Planarian mitochondria sequence heterogeneity: relationships between the type of cytochrome c oxidase subunit I gene sequence, karyotype and genital organ.

Freshwater planarians Dugesia japonica from three localities were examined for cytochrome c oxidase subunit I (COI) gene sequence, karyotype and the presence of genital organ. The planarians from Mt Fujiwara in Japan were composed of two different groups; one revealed inter- and intraindividual COI gene heterogeneity, while another revealed no sequence heterogeneity. The sequence in planarians from Mt Alishan in Taiwan was homogeneous, while that from the Kenting National Park in Taiwan revealed a considerable heterogeneity. All the planarians having the homogeneous gene sequences carry the 2X karyotype and many of them had genital organs. These are assumed to belong to the sexual lineage. In contrast, almost all planarians having heterogeneous sequences carry the karyotype of either 3X plus 2X (mixoploid) or 3X, and all of them lack genital organs. These lineages are assumed to be asexual. The heterogeneity of COI gene sequences in the presumed asexual lineages would have resulted from an accumulation of mutations by repeated asexual reproduction.

Animals↗
PubMed1996

Molecular evidence for Acanthocephala as a subtaxon of Rotifera.

Rotifers are free-living animals usually smaller than 1 mm that possess a characteristic wheel organ. Acanthocephalans (thorny-headed worms) are larger endoparasitic animals that use vertebrates and arthropods to complete their life cycle. The taxa Acanthocephala and Rotifera are considered separate phyla, often within the taxon Aschelminthes. We have reexamined the relationship between Rotifera and Acanthocephala using 18S rRNA gene sequences. Our results conclusively show that Acanthocephala is the sister group of the rotifer class Bdelloidea. Rotifera was nonmonophyletic in all molecular analyses, which supports the hypothesis that the Acanthocephala represent a taxon within the phylum Rotifera and not a separate phylum. These results agree with a previous cladistic study of morphological characters.

Acanthocephala↗
PubMed1996

Molecular genetic approaches to parasite identification: their value in diagnostic parasitology and systematics.

A wide range of approaches is available to parasitologists to aid in specific parasite identification and to formulate phylogenetic relationships. This review emphasises the usefulness of molecular genetic techniques, especially DNA-based procedures, in addressing problems of identification, characterisation and phylogeny of parasites. It should be stressed that an understanding of the various DNA approaches, techniques and target genes most likely to be effective in addressing key issues in diagnostic parasitology and systematics is still developing. Nevertheless, DNA methods clearly have great potential with regard to specificity and sensitivity, and applications will increase further with technological advance. Indeed, because of the minimal requirements for material, PCR-based methods especially should prove of immense value in future studies with parasites.

Animals↗
PubMed1995

A molecular phylogeny of the genus Echinococcus.

Three nucleotide data sets, two mitochondrial (COI and ND1) and one nuclear (ribosomal ITS1), have been investigated in order to resolve relationships among species and strains of the genus Echinococcus. The data have some unusual properties in that mitochondrial heteroplasmy was detected in one strain of E. granulosus, and more than one class of ITS1 sequence variant can occur in a single isolate. The data failed to support the hypothesis that E. granulosus, as it is currently viewed, is a single valid species. Rather, the strains of E. granulosus seem to comprise at least three evolutionarily diverse groups, the sheep strain group, bovine strain group and horse strain group. Molecular distances between them are comparable to, or greater than, molecular evolutionary distances observed between recognized species. The affinities of the cervid strain of E. granulosus are unclear because of ambiguous data, but this strain does not appear to be ancestral to others. E. multilocularis may not be distinct from E. granulosus. However, the remaining two species. E. vogeli and E. oligarthrus appear distinct and rather distant from the first two. Based on the results presented here, taxonomic revision of the genus is clearly warranted.

Animals↗
PubMed1996

Genetic markers in ribosomal DNA for hookworm identification.

Polymerase chain reaction-linked restriction fragment length polymorphism (PCR-RFLP) analysis of ribosomal (r) DNA was conducted on Uncinaria stenocephala, Ancylostoma caninum, A. tubaeforme and A. ceylanicum. The rDNA region spanning the first and second internal transcribed spacers (ITS1 and ITS2) plus the 5.8S (ITS+) gene was amplified by PCR from each of the species, digested separately with 9 restriction endonucleases and the fragments separated by agarose gel electrophoresis. Characteristic PCR-RFLP patterns were produced for each morphologically defined species using some of the endonucleases. The present study demonstrated that the ITS+ provides genetic markers for the delineation of each species examined and suggests that this region of rDNA will be useful for the identification of other hookworms from a range of hosts. The results are likely to have important implications for studying the genetic structure of hookworm populations, the systematics and the epidemiology of hookworm infections.

Ancylostomatoidea↗
PubMed1997

Species-specific polymerase chain reaction for the differentiation of larvae from Dictyocaulus viviparus and Dictyocaulus eckerti.

Using substantial interspecific differences between the second internal transcribed spacer (ITS2) region within the rDNA gene of Dictyocaulus eckerti and Dictyocaulus viviparus a species-specific PCR was developed to distinguish between lungworm larvae of the two species from fallow deer and cattle. It was found that the method of DNA extraction was crucial for the sensitivity of the PCR. With serial dilutions of DNA extracted from 10,000 larvae the ITS2 fragment could be amplified from all dilutions down to a calculated amount of DNA equivalent to one larva. Using lower numbers of larvae, DNA from at least 100 larvae was necessary for a successful amplification. From this extraction a species-specific polymerase chain reaction (PCR) product was generated with a calculated amount of DNA equivalent to 33 larvae, whereas amplification of further diluted DNA was not successful. However, in a direct PCR single larvae could be detected after direct PCR amplification without preceding DNA extraction.

Animals↗