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: The occurrence of P. hermaphrodita in North America has regulatory implications for potential biological control strategies against non-native gastropod species that are pests in ornamental and agricultural cultivation on this continent.
Abstract: Phasmarhabditis hermaphrodita is reported for the first time in North America from cadavers of the invasive slug species Deroceras reticulatum, D. laeve and Lehmannia valentiana collected from three different locations in California, USA. Four isolates were identified using combined morphology, morphometrics and molecular sequence data for complete internal transcribed spacer (ITS-1, 5.8S, ITS-2), D2-D3 expansion segments of the large subunit (LSU or 28S) and nearly complete small subunit (SSU or 18S) ribosomal DNA. Extremely low sequence variations in the COI gene of the mitochondria were observed among US isolates as well as between US isolates and the two UK sequences. The occurrence of P. hermaphrodita in North America has regulatory implications for potential biological control strategies against non-native gastropod species that are pests in ornamental and agricultural cultivation on this continent. The D2-D3 sequence of the LSU rDNA is new for the species.
47 citations
Cites background from "Molecular phylogenetics and diagnos..."
...Molecular sequence data for complete internal transcribed spacer (ITS-1, 5.8S, ITS-2), D2-D3 expansion segment of the LSU and nearly complete SSU ribosomal DNA, as well as the COI gene of the mitochondria further corroborated the identity of our isolates based on morphology and morphometrics....
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...…et al., 1998; Tandingan De Ley et al., 2002); N93 forward (5′-TTGAACCGGGTAAAAGTCG-3′) and N94 reverse (5′-TTAGTTTCTTTTCCTCCGCT-3′) primers for ITS (Nadler et al., 2005); and forward primer COIF1 (5′-CCTACTATGATTGGTGGTTTTGGTAATTG-3′) and reverse primer COI-R2 (5-GTAGCAGCAGTAAAA TAAGCACG-3′)…...
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...2-3 μl of the genomic template DNA was used in a 25 μl PCR reaction using Illustra PuReTaq Ready-To-Go™ PCR beads (GE Healthcare) under the same PCR conditions and using the same amplification and sequencing primers previously described for D2-D3 and SSU (Blaxter et al., 1998; Tandingan De Ley et al., 2002); N93 forward (5′-TTGAACCGGGTAAAAGTCG-3′) and N94 reverse (5′-TTAGTTTCTTTTCCTCCGCT-3′) primers for ITS (Nadler et al., 2005); and forward primer COIF1 (5′-CCTACTATGATTGGTGGTTTTGGTAATTG-3′) and reverse primer COI-R2 (5-GTAGCAGCAGTAAAA TAAGCACG-3′) for mtCOI (Kanzaki & Futai, 2002)....
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...Four isolates were identified using combined morphology, morphometrics and molecular sequence data for complete internal transcribed spacer (ITS-1, 5.8S, ITS-2), D2-D3 expansion segments of the large subunit (LSU or 28S) and nearly complete small subunit (SSU or 18S) ribosomal DNA....
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..., 2002); N93 forward (5′-TTGAACCGGGTAAAAGTCG-3′) and N94 reverse (5′-TTAGTTTCTTTTCCTCCGCT-3′) primers for ITS (Nadler et al., 2005); and forward primer COIF1 (5′-CCTACTATGATTGGTGGTTTTGGTAATTG-3′) and reverse primer COI-R2 (5-GTAGCAGCAGTAAAA TAAGCACG-3′) for mtCOI (Kanzaki & Futai, 2002)....
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TL;DR: First record of the presence of A. pegreffii is reported from Turkish coasts of Aegean and Mediterranean Sea, suggesting a high threat of anisakiasis or allergies for Turkish populations who consume fish originating in these regions.
46 citations
TL;DR: An overall concordant tree topology showed C. bioccai n.
Abstract: Contracaecum bioccai n. sp. is described from the brown pelican Pelecanus occidentalis (L.) in northern Colombia (Totumo Marsh) based on 20 enzyme loci studied using multilocus allozyme electrophoresis. Moreover, genetic relationships between the new taxon and related congeners are presented based on allozyme data-sets and sequence analyses (519 bp) of the mtDNA-cox2 gene. Fixed allele differences were found at some of the allozyme loci analysed in comparison with other Contracaecum spp. from pelicans and cormorants [i.e. the sibling species of the C. rudolphii Hartwich, 1964 complex, C. septentrionale Kreis, 1955, C. micropapillatum (Stossich, 1890), C. microcephalum (Rudolphi, 1809) and C. pelagicum Johnston & Mawson, 1942]. The genetic distance, at the allozyme level, between C. bioccai n. sp. and its congeners ranged from D ( Nei ) = 0.80 versus C. septentrionale to D ( Nei ) = 1.40 versus C. micropapillatum. The genetic distance at the mtDNA cox-2 level ranged, on average, from K-2P = 0.12 versus the C. rudolphii species complex to K-2P = 0.15 versus C. micropapillatum. An overall concordant tree topology, obtained from UPGMA and NJ tree analyses inferred from allozyme data, as well as from MP, UPGMA and NJ inferred from mtDNA-cox2 sequence analysis, showed C. bioccai n. sp. as a separated lineage to the other Contracaecum spp. A concordant result was also obtained by PCA analysis based on both the allozyme and mtDNA cox-2 data-sets. All of the tree topologies, derived from the phylogenetic analysis inferred from both allozymes and mtDNA data-sets, were in substantial agreement and depicted C. bioccai as closely related to the sibling species of the C. rudolphii complex (C. rudolphii A and C. rudolphii B) and C. septentrionale. Morphological analysis and a differential diagnosis based on male specimens of C. bioccai, which had been genetically characterised by both allozyme markers and mtDNA sequences analysis with respect to morphologically related congeners, enabled the detection of differences in a numbers of characters, including spicule length, the morphology of the distal end of the spicule and the distribution patterns of the distal caudal papillae.
46 citations
Cites background or methods from "Molecular phylogenetics and diagnos..."
...Moreover, other genetic/ molecular markers applied to the systematics of Contracaecum have enabled the study of genetic relationships between some Contracaecum spp. from fish-eating birds and those which are parasites of seals (Nascetti et al., 1990; Nadler et al., 2000, 2005)....
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...…al., 1993) from Halichoerus grypus (Genbank EF122211) were included as outgroup to root the Contracaecum phylogenetic trees, based on the relationships of Contracaecum (s. s.). and Pseudoterranova spp., previously demonstrated in ribosomal and mitochondrial DNA analyses (Nadler et al., 2000, 2005)....
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TL;DR: Panmictic distribution of the larval Anisakis spp.
Abstract: Members of the Anisakidae are known to infect over 200 pelagic fish species and have been frequently used as biological tags to identify fish populations. Despite information on the global distribution of Anisakis species, there is little information on the genetic diversity and population structure of this genus, which could be useful in assessing the stock structure of their fish hosts. From 2005 through 2008, 148 larval anisakids were recovered from Pacific sardine (Sardinops sagax) in the California Current upwelling zone and were genetically sequenced. Sardines were captured off Vancouver Island, British Columbia in the north to San Diego, California in the south. Three species, Anisakis pegreffii, Anisakis simplex 'C', and Anisakis simplex s.s., were identified with the use of sequences from the internal transcribed spacers (ITS1 and ITS2) and the 5.8s subunit of the nuclear ribosomal DNA. The degree of nematode population structure was assessed with the use of the cytochrome c oxidase 2 (cox2) mitochondrial DNA gene. All 3 Anisakis species were distributed throughout the study region from 32u Nt o 50uN latitude. There was no association between sardine length and either nematode infection intensity or Anisakis species recovered. Larval Anisakis species and mitochondrial haplotype distributions from both parsimony networks and analyses of molecular variance revealed a panmictic distribution of these parasites, which infect sardines throughout the California Current ecosystem. Panmictic distribution of the larval Anisakis spp. populations may be a result of the presumed migratory pathways of the intermediate host (the Pacific sardine), moving into the northern portion of the California Current in summer and returning to the southern portion to overwinter and spawn in spring. However, the wider geographic range of paratenic (large piscine predators), and final hosts (cetaceans) can also explain the observed distribution pattern. As a result, the recovery of 3 Anisakis species and a panmictic distribution of their haplotypes could not be used to confirm or deny the presence of population subdivision of Pacific sardines in the California Current system.
45 citations
Additional excerpts
...…ribosomal DNA (rDNA) (hereafter referred to as ITS markers) to identify larval Anisakis nematodes to species genetically at 2 diagnostic nucleotide sites (Abollo et al., 2003; Nadler et al., 2005; Abe, 2008) with the use of the forward primer 93 and the reverse primer 94 (Nadler et al., 2005)....
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TL;DR: It is revealed that the life-cycle of P. cattani involves mainly demersal and benthic organisms, with a marked preference by large-sized benthophagous fish, whereas few host species harbour these larvae in Argentina where they show low levels of parasitism.
45 citations
References
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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