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Journal ArticleDOI

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.

AbstractIndividual 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.

Topics: Anisakis (60%), Eumetopias jubatus (51%)

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Citations
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Book ChapterDOI
TL;DR: Examples in which anisakid nematodes recognized genetically at the species level in definitive and intermediate/paratenic hosts from various geographical areas of the Boreal and Austral regions and their infection levels have been used as biological indicators of fish stocks and food-web integrity in areas at high versus low levels of habitat disturbance are presented.
Abstract: The application of molecular systematics to the anisakid nematodes of the genera Anisakis, Pseudoterranova and Contracaecum, parasites of aquatic organisms, over the last two decades, has advanced the understanding of their systematics, taxonomy, ecology and phylogeny substantially Here the results of this effort on this group of species from the early genetic works to the current status of their revised taxonomy, ecology and evolutionary aspects are reviewed for each of three parasitic groups It has been shown that many anisakid morphospecies of Anisakis, Contracaecum and Pseudoterranova include a certain number of sibling species Molecular genetic markers provided a rapid, precise means to screen and identify several species that serve as definitive and intermediate and or/paratenic hosts of the so far genetically characterized species Patterns of differential distribution of anisakid nematodes in various definitive and intermediate hosts are presented Differences in the life history of related species can be due both to differential host-parasite co-adaptation and co-evolution, and/or to interspecific competition, that can reduce the range of potential hosts in sympatric conditions Phylogenetic hypotheses attempted for anisakid nematodes and the possible evolutionary scenarios that have been proposed inferred from molecular data, also with respect to the phylogeny of their hosts are presented for the parasite-host associations Anisakis-cetaceans and Contracaecum-pinnipeds, showing that codivergence and host-switching events could have accompanied the evolution of these groups of parasites Finally, examples in which anisakid nematodes recognized genetically at the species level in definitive and intermediate/paratenic hosts from various geographical areas of the Boreal and Austral regions and their infection levels have been used as biological indicators of fish stocks and food-web integrity in areas at high versus low levels of habitat disturbance (pollution, overfishing, by-catch) are presented

377 citations


Cites background or methods or result from "Molecular phylogenetics and diagnos..."

  • ...pegreffii is also well supported by a phylogenetic analysis inferred from ITS rDNA sequence data sets (Nadler et al., 2005)....

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  • ...Codes: AN: Antarctica; BE: Bering Sea; BS: Barents Sea; FI: Faeroe Islands; JA: Japan Sea; LS: Labrador Sea; NEA: North-East Atlantic; NEP: North-East Pacific; NWA: NorthWest Atlantic; NWP: North-West Pacific; SEP: South-East Pacific (Chilean coast) (data from George-Nascimento and Llanos, 1995; George-Nascimento and Urrutia, 2000; Mattiucci et al., 1998; Nadler et al., 2005; Paggi et al., 1991, 1998c; Zhu et al., 2002)....

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  • ...) is widespread between 35 N and the Arctic Circle; it is present in both the western and eastern Atlantic and Pacific Oceans (Abe et al., 2005, 2006; Abollo et al., 2001; Mattiucci et al., 1997, 1998; Nadler et al., 2005; Paggi et al., 1998a; Umehara et al., 2006, 2008) (Fig....

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  • ...Parsimony and ML analyses indicated that the Raphidascarididae, Contracaecum plus Phocascaris, and the Anisakinae (here considering only Pseudoterranova and Anisakis) are each monophyletic, the latter two groups with consistently strong bootstrap support at MP and ML analyses (Nadler et al., 2005)....

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  • ...A congruent result was inferred from the ITS rDNA sequence analysis (Nadler et al., 2005) (see Section 4....

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Journal ArticleDOI
TL;DR: The morphological and morphometric analyses revealed the presence of morphological features that differed among the 3 biological species, and the tree topologies support the finding that A. simplex (s.s.), A. pegreffii, and A. berlandi n.
Abstract: Numerous specimens of the 3 sibling species of the Anisakis simplex species complex (A. pegreffii, A. simplex (senso stricto)), and A. simplex sp. C) recovered from cetacean species stranded within the known geographical ranges of these nematodes were studied morphologically and genetically. The genetic characterization was performed on diagnostic allozymes and sequences analysis of nuclear (internal transcribed spacer [ITS] of ribosomal [r]DNA) and mitochondrial (mitochondrial [mt]DNA cox2 and rrnS) genes. These markers showed (1) the occurrence of sympatry of the 2 sibling species A. pegreffii and A. simplex sp. C in the same individual host, the pilot whale, Globicephala melas Traill, from New Zealand waters; (2) the identification of specimens of A. pegreffii in the striped dolphin, Stenella coeruleoalba (Meyen), from the Mediterranean Sea; and (3) the presence of A. simplex (s.s.) in the pilot whale and the minke whale, Balaenoptera acutorostrata Lacepede, from the northeastern Atlantic wa...

132 citations


Cites background from "Molecular phylogenetics and diagnos..."

  • ...…have confirmed the existence of these 3 cryptic species by using phylogenetic analyses of DNA sequences of mitochondrial (mitochondrial [mt]DNA cox2) and nuclear (internal transcribed spacer [ITS] of ribosomal [r]DNA) genes (Nadler et al., 2005; Mattiucci et al., 2009; Cavallero et al., 2011)....

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Journal ArticleDOI
TL;DR: It is suggested that anisakiasis in Japan is mainly caused by A. simplex sensu stricto because it penetrates the muscle of the fish at a higher rate than A. pegreffii.
Abstract: It has been reported that nearly all cases of anisakiasis in Japan are caused by Anisakis simplex sensu stricto. To elucidate this further, we investigated the presence of Anisakis type I larvae and Pseudoterranova decipiens in 218 Scomber japonicus fish collected from the seas of Japan. Anisakis type I larvae were detected in 74.3% (162/218) of the fish, and 99.8% of the Anisakis type I larvae comprised A. simplex sensu stricto and Anisakis pegreffii. Molecular identification techniques were used for 7.5% (360/4806) of the Anisakis type I larvae. The larvae found in the fish of the Pacific stock (the Pacific coast of Japan) and the Tsushima Warm Current stock (the East China Sea and the Sea of Japan) were primarily A. simplex sensu stricto and A. pegreffii, respectively. In addition, for the first time in Japan, Anisakis simplex C and Anisakis ziphidarum were detected in the fish of the Pacific stock. The average number of A. pegreffii and A. simplex sensu stricto larvae per fish was 47 and 6, respectively. However, the average number (0.61 larvae) of A. simplex sensu stricto in the muscle per fish was 12 times the average number (0.05 larvae) of A. pegreffii. When fish on the purchased day were compared with those held at 4 degrees C and 20 degrees C for 20h, the penetration rates (ratio of the number of larvae detected in the muscle to the total number of larvae detected) of A. pegreffii and A. simplex sensu stricto were as high as 1.8% and 5.8%, respectively. In conclusion, we suggest that anisakiasis in Japan is mainly caused by A. simplex sensu stricto because it penetrates the muscle of the fish at a higher rate than A. pegreffii.

126 citations


Journal ArticleDOI
TL;DR: This is the first instance of human intestinal anisakiasis diagnosed using PCR of DNA purified from a fixed eosinophilic granuloma embedded in paraffin, reinforcing the pathological significance of the species A. pegreffii to humans.
Abstract: Anisakiasis is an important fish-borne zoonosis provoked by larval stages of nematodes belonging to the genus Anisakis. The detection and identification of human infections is difficult. This is due to: a) the low specificity of the clinical features and symptomatology related to human infections; b) the paucity of diagnostic features of larvae found in granulomatous lesions characteristic of "invasive anisakiasis"; and c) the lack morphological characters diagnostic at the specific level when larvae of Anisakis are detected. Thus, molecular-based diagnostic approaches are warranted. We have developed a PCR method that amplifies the DNA of Anisakis spp. in fixed paraffin-embedded tissues. This method was applied to a granuloma removed from a human case of intestinal anisakiasis in Italy. Specific primers of the mtDNA cox2 gene were used and sequence analysis was performed according to the procedures already established for species of Anisakis. The sequence obtained (629 bp) was compared with those of the other species of Anisakis which have so far been genetically characterized and with sequences obtained from larval stages of Anisakis collected from the Mediterranean fish Engraulis encrasicolus. This enabled the genetic identification of the larva in the human tissue as A. pegreffii. This is the first instance of human intestinal anisakiasis diagnosed using PCR of DNA purified from a fixed eosinophilic granuloma embedded in paraffin. The case of human anisakiasis presented reinforces the pathological significance of the species A. pegreffii to humans. The molecular/genetic methodological approach based on mtDNA cox2 sequence analysis, described here, can allow easy and rapid identification of Anisakis spp. in formalin-fixed and paraffin embedded tissues removed from cases of either gastric or intestinal human anisakiasis.

116 citations


Cites background from "Molecular phylogenetics and diagnos..."

  • ...The systematics of the species of Anisakis has been clarified in recent decades by the application of such molecular methodologies [1,18-34]....

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Book ChapterDOI
TL;DR: This review addresses the biodiversity, biology, distribution, ecology, epidemiology, and consumer health significance of the so far known species of Anisakis, both in their natural hosts and in human accidental host populations, worldwide.
Abstract: This review addresses the biodiversity, biology, distribution, ecology, epidemiology, and consumer health significance of the so far known species of Anisakis, both in their natural hosts and in human accidental host populations, worldwide. These key aspects of the Anisakis species' biology are highlighted, since we consider them as main driving forces behind which most of the research in this field has been carried out over the past decade. From a public health perspective, the human disease caused by Anisakis species (anisakiasis) appears to be considerably underreported and underestimated in many countries or regions around the globe. Indeed, when considering the importance of marine fish species as part of the everyday diet in many coastal communities around the globe, there still exist significant knowledge gaps as to local epidemiological and ecological drivers of the transmission of Anisakis spp. to humans. We further identify some key knowledge gaps related to Anisakis species epidemiology in both natural and accidental hosts, to be filled in light of new 'omic' technologies yet to be fully developed. Moreover, we suggest that future Anisakis research takes a 'holistic' approach by integrating genetic, ecological, immunobiological, and environmental factors, thus allowing proper assessment of the epidemiology of Anisakis spp. in their natural hosts, in human populations, and in the marine ecosystem, in both space and time.

100 citations


References
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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.

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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?

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Journal ArticleDOI
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.)

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Journal ArticleDOI
15 Apr 1961-Sarsia
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.

346 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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Journal ArticleDOI
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.
Abstract: Polymerase-chain-reaction-based restriction fragment length polymorphism analysis was performed to establish genetic markers in rDNA, for the identification of the three sibling species of the Anisakis simplex complex and morphologically differentiated Anisakis species, i.e. Anisakis physeteris, Anisakis schupakovi, Anisakis typica and Anisakis ziphidarum. Different restriction patterns were found between A. simplex sensu stricto and Anisakis pegreffii with two of the restriction endonucleases used (HinfI and TaqI), between A. simplex sensu stricto and A. simplex C with one endonuclease (HhaI), and between A. simplex C and Aniskis pegreffii with three endonucleases (HhaI, HinfI and TaqI), while no variation in patterns was detected among individuals within each species. The species A. physeteris, A. schupakovi, A. typica and A. ziphidarum were found to be different from each other and different from the three sibling species of the A. simplex complex by distinct fragments using 10-12 of the endonucleases tested. 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.

275 citations