Molecular phylogenetics and diagnosis of Anisakis, Pseudoterranova, and Contracaecum from northern Pacific marine mammals.
01 Dec 2005-Journal of Parasitology (American Society of Parasitologists)-Vol. 91, Iss: 6, pp 1413-1429
TL;DR: Phylogenetic analysis of LSU sequences revealed strong support for the monophyly of Anisakinae, Contracaecum plus Phocascaris, Pseudoterranova, and Anisakis, which is primarily consistent with previously published phenograms based on multilocus electrophoretic data.
Abstract: Individual specimens of Anisakis, Pseudoterranova, and Contracaecum collected from marine mammals inhabiting northern Pacific waters were used for comparative diagnostic and molecular phylogenetic analyses. Forty-eight new sequences were obtained for this study of 14 Anisakis taxa, 8 Pseudoterranova taxa, 4 Contracaecum taxa, and 4 outgroup species. Partial 28S (LSU) and complete internal transcribed spacer (ITS-1, 5.8S, ITS-2) ribosomal DNA was amplified by the polymerase chain reaction and sequenced. Sequences of ITS indicated that Pseudoterranova specimens from Zalophus californianus (California sea lion), Mirounga angustirostris (northern elephant seal), Phoca vitulina (harbor seal), Enhydra lutris (sea otter), and Eumetopias jubatus (Steller's sea lion) exactly matched P. decipiens s. str., extending the host and geographic range of this species. Anisakis from northern Pacific marine mammals were most closely related to members of the A. simplex species complex. Comparison of Anisakis ITS sequences diagnosed the presence of A. simplex C in 2 M. angustirostris hosts, which is a new host record. Anisakis specimens from Phocoena phocoena (harbor porpoise), Lissodelphis borealis (Pacific rightwhale porpoise), and E. jubatus included 3 ITS sequences that did not match any known species. Contracaecum adults obtained from Z. californianus were most closely related to C. ogmorhini s.l. and C. rudolphii, but ITS sequences of these Contracaecum specimens did not match C. ogmorhini s. str. or C. margolisi. These novel Anisakis and Contracaecum ITS sequences may represent previously uncharacterized species. Phylogenetic analysis of LSU sequences revealed strong support for the monophyly of Anisakinae, Contracaecum plus Phocascaris, Pseudoterranova, and Anisakis. Phylogenetic trees inferred from ITS sequences yielded robustly supported relationships for Pseudoterranova and Anisakis species that are primarily consistent with previously published phenograms based on multilocus electrophoretic data.
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TL;DR: This is the first report of P. papillosa from the African continent and of its virulence against D. panormitanum, and the nematode was identified using a combination of morphological, morphometric, molecular and phylogenetic techniques.
Abstract: Worldwide interest in Phasmarhabditis originates from the successful commercialisation of P. hermaphrodita as a biological control agent against molluscs in Europe. To date, P. hermaphrodita has not been isolated from South Africa and, therefore, the formulated product may not be sold locally. During a survey for mollusc-associated nematodes, P. papillosa was dissected from the slug, Deroceras reticulatum , collected from George, South Africa. The nematode was identified using a combination of morphological, morphometric, molecular and phylogenetic techniques. Virulence tests were conducted which demonstrated that P. papillosa caused significant mortality to the European invasive slug Deroceras panormitanum . Additional data are provided in the morphometrics of the infective juvenile and in the molecular identification, using the mitochondrial cytochrome c oxidase subunit I ( cox1 ) gene. This is the first report of P. papillosa from the African continent and of its virulence against D. panormitanum .
26 citations
TL;DR: The sequence analysis confirmed the hybridisation in the 85% of the larvae recognised as hybrid forms by PCR- RFLP, suggesting this form as the product of natural interspecific recombination due to the presence of sympatry areas.
26 citations
TL;DR: All anisakid larvae in the present study showed the most typical features of the genus Contracaecum, and sequencing and sequence and phylogenetic analyses showed that the present nematode larvae are likely belonging to C. multipapillatum.
Abstract: Nematodes of the family Anisakidae are parasites of many fishes and aquatic invertebrates which act as intermediate or paratenic hosts, while mammals and fish-eating birds are definitive hosts. Infective L3 larvae may be incidentally taken by human through eating raw or undercooked fish meat, causing anisakidosis. The main purpose of this study is to provide a basis for the future investigations to discover the genetic diversity of this widely distributed parasite nematodes of fishes and fish eating animals and their effect on fisheries and public health in Egypt and worldwide. One thousand, one hundred and fifteen specimens belong to nine fish species were collected from Lake Nasser, Egypt, and examined for infection with Anisakid larvae. Four fish species (Oreochromis niloticus, Tilapia galilaea, Lates niloticus, and Hydrocynus forskahlii) were found infected with third stage larvae of Contracaecum spp. No other Anisakid nematodes were detected. Larvae were found in the body cavity adhering to mesenteries by a thin membrane, except in Oreochromis niloticus and Tilapia galilaea were found free in branchial chambers. The highest prevalence was recorded in L. niloticus (100%) and H. forskahlii (82%). The mean intensity of infections were 0.17–4.12 and 5.1–10.3 in L. niloticus and H. forskahlii respectively. For further identification, the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA from isolated larvae (n = 54) were amplified by PCR, followed by single-strand conformation polymorphism (SSCP) analysis which revealed five possible profiles. Light and scanning electron microscope studies revealed that all anisakid larvae in the present study showed the most typical features of the genus Contracaecum. The sequencing (n = 28) and sequence and phylogenetic analyses showed that the present nematode larvae are likely belonging to C. multipapillatum.
25 citations
TL;DR: The stargraph method was applied to visualize observed regional differences using grouper parasites as biological indicators for the sampled coastal ecosystems at both sampling sites and revealed 21 different parasite species, and 14 new host and locality records.
Abstract: Sixty Epinephelus areolatus were examined for metazoan fish parasites in Indonesia, off Segara Anakan lagoon, Java and in Balinese waters. The study revealed 21 different parasite species, and 14 new host and locality records. The anisakid nematodes Anisakis typica and, for the first time in Indonesia, Anisakis sp. HC-2005 were identified by using molecular methods. Ecological parameters were calculated for both sites off the anthropogenically influenced Segara Anakan lagoon and the relatively undisturbed reference site at the southern Balinese coast. The fish from Segara Anakan demonstrated a significantly higher enzymatic activity (Hepatosomatic index) and a significantly reduced number of heteroxenous gut helminths (e.g. the digenean Didymodiclinus sp., the nematode Raphidascaris sp. and the acanthocephalan Serrasentis sagittifer). Other regional differences for E. areolatus included ecto-/endoparasite ratio, endoparasite diversity, the parasite species composition and prevalence of infection of the respective parasite species. We applied the stargraph method to visualize observed regional differences using grouper parasites as biological indicators for the sampled coastal ecosystems at both sampling sites.
25 citations
Cites background from "Molecular phylogenetics and diagnos..."
...Recently, this common tropical nematode species has also been identified genetically from the South-west Atlantic (Brazil; Nadler et al., 2005), Northwest Atlantic (Florida; Mattiucci et al., 2005) and the Mediterranean (North Africa; Farjallah et al., 2008)....
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...Recently, this common tropical nematode species has also been identified genetically from the South-west Atlantic (Brazil; Nadler et al., 2005), Northwest Atlantic (Florida; Mattiucci et al....
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TL;DR: Using high-fidelity polymerase chain reaction to amplify a large portion of the ribosomal gene complex of four pinworm species commonly found in lab rodents and rabbits, real-time molecular beacon assays for pinworm detection are developed and compared with traditional methods such as perianal tape testing, fecal flotation and direct examination of intestinal content.
Abstract: Though pinworm infestation has been prevalent since the early years of laboratory animal medicine, the genomes of these parasites have not yet been sequenced. The authors used high-fidelity polymerase chain reaction to amplify a large portion of the ribosomal gene complex of four pinworm species commonly found in lab rodents and rabbits (Aspiculuris tetraptera, Passalurus ambiguus, Syphacia muris and Syphacia obvelata). They determined DNA sequences for these complexes and carried out phylogenetic analysis. Using this information, the authors developed real-time molecular beacon assays for pinworm detection, comparing the new diagnostic approach with traditional methods such as perianal tape testing, fecal flotation and direct examination of intestinal content.
24 citations
References
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TL;DR: The program MODELTEST uses log likelihood scores to establish the model of DNA evolution that best fits the data.
Abstract: Summary: The program MODELTEST uses log likelihood scores to establish the model of DNA evolution that best fits the data. Availability: The MODELTEST package, including the source code and some documentation is available at http://bioag.byu.edu/zoology/crandall―lab/modeltest.html. Contact: dp47@email.byu.edu.
20,105 citations
"Molecular phylogenetics and diagnos..." refers methods in this paper
...Modeltest Version 3.06 (Posada and Crandall, 1998) was used to compare the fit of nucleotide substitution models for datasets using the Akaike information criterion....
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TL;DR: The effects on phylogenetic accuracy of adding characters and/or taxa were explored using data generated by computer simulation using a four-taxon tree representing a difficult phylogenetic problem with an extreme situation of long branch attraction.
Abstract: The effects on phylogenetic accuracy of adding characters and/or taxa were explored using data generated by computer simulation. The conditions of this study were constrained but allowed for systematic investigation of certain parameters. The starting point for the study was a four-taxon tree in the ``Felsenstein zone,'' representing a dif® cult phylogenetic problem with an extreme situation of long branch attraction. Taxa were added sequentially to this tree in a manner speci® cally designed to break up the long branches, and for each tree data matrices of different sizes were simulated. Phylogenetic trees were reconstructed from these data using the criteria of parsimony and maximum likelihood. Phylogenetic accuracy was measured in three ways: (1) proportion of trees that are completely correct, (2) proportion of correctly reconstructed branches in all trees, and (3) proportion of trees in which the original four-taxon statement is correctly reconstructed. Accuracy improved dramatically with the addition of taxa and much more slowly with the addition of characters. If taxa can be added to break up long branches, it is much more preferable to add taxa than characters. (Long branch attraction; parsimony; phylogenetic recon- struction; simulation; taxon sampling.)
687 citations
01 Jan 1998
TL;DR: In this paper, the effects of adding characters and/or taxa on phylogenetic accuracy were explored using data generated by computer simulation using a four-taxon tree in the "Felsenstein zone/7".
Abstract: The effects on phylogenetic accuracy of adding characters and/or taxa were explored using data generated by computer simulation. The conditions of this study were constrained but allowed for systematic investigation of certain parameters. The starting point for the study was a four-taxon tree in the "Felsenstein zone/7 representing a difficult phylogenetic problem with an extreme situation of long branch attraction. Taxa were added sequentially to this tree in a manner specifically designed to break up the long branches, and for each tree data matrices of different sizes were simulated. Phylogenetic trees were reconstructed from these data using the criteria of parsimony and maximum likelihood. Phylogenetic accuracy was measured in three ways: (1) proportion of trees that are completely correct, (2) proportion of correctly reconstructed branches in all trees, and (3) proportion of trees in which the original four-taxon statement is correctly reconstructed. Accuracy improved dramatically with the addition of taxa and much more slowly with the addition of characters. If taxa can be added to break up long branches, it is much more preferable to add taxa than characters. (Long branch attraction; parsimony; phylogenetic recon? struction; simulation; taxon sampling.) It is obvious that the successful recon? struction of phylogenetic relationships re? quires some amount of data sampling from relevant taxa and informative char? acters. Far less clear, however, is how much of each data type is required, and whether one of those sources of data has a greater impact on accuracy than the other. Given limited time and resources, it is important to explore the costs and benefits to phylo? genetic accuracy of adding taxa versus in? creasing the number of characters. For ex? ample, given that one has sufficient time and resources to sequence 10 kilobases (kb) of DNA, would it be better to se? quence, say, 2.5 kb from each of 4 taxa, or 1 kb from each of 10, or 0.25 kb from each of 40?
676 citations
TL;DR: More than 260 specimens of fish, belonging to 64 species, have been searched for nematode parasites and 9 species of fish were found to be free of nematodes.
Abstract: The present paper deals with nematode parasites from some Norwegian marine fishes. The major part of the material was collected personally from fishes caught in the fjords near Bergen and Tromso; the minor part of the material was obtained from the zoological museums in Oslo and Bergen and from other sources. 260 specimens of fish, belonging to 64 species, have been searched for nematode parasites. 9 species of fish were found to be free of nematodes.
373 citations
"Molecular phylogenetics and diagnos..." refers background in this paper
...…A. simplex s. str.) A. simplex C); these represent the 3 known species within the A. simplex species complex and are characterized (along with A. typica and A. ziphidarum) by having type I larvae sensu Berland (1961), which may represent the apomorphic (derived) state within An- NADLER ET AL....
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TL;DR: The polymorphisms obtained by restriction fragment length polymorphisms have provided a new set of genetic markers for the accurate identification of sibling species and morphospecies.
297 citations