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: 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
402 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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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...
166 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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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.
158 citations
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.
142 citations
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.
128 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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References
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TL;DR: The helminth fauna of the gastrointestinal tract and abdominal cavity of cutlass fish, Trichiurus lepturus L., off the Taiwanese coast of the north-western Pacific was investigated and a PCR-based restriction fragment length polymorphism method was established for the unequivocal delineation of the four species.
Abstract: The helminth fauna of the gastrointestinal tract and abdominal cavity of cutlass fish, Trichiurus lepturus L., off the Taiwanese coast of the north-western Pacific was investigated. The following helminths were found: (1) nematodes—Anisakis simplex, Hysterothylacium aduncum, Porrocaecum decipiens, Raphidascaris trichiuri; (2) digeneans—adult Lecithochirium trichiuri; and (3) cestodes—plerocercoids of Proteocephalus spp. The third-stage larvae of these four anisakid nematodes were characterized genetically using a molecular approach. The nuclear ribosomal DNA region spanning the first internal transcribed spacer (ITS-1), the 5.8S gene and the second internal transcribed spacer (ITS-2) was amplified and sequenced. Based on the sequence differences, a PCR-based restriction fragment length polymorphism method was established for the unequivocal delineation of the four species. Phylogenetic analysis showed that H. aduncum clustered with P. decipiens, whereas A. simplex was not closely related to these according to the nucleotide sequences of all rDNA.
50 citations
"Molecular phylogenetics and diagnos..." refers background or methods in this paper
...Phylogenetic support for Anisakidae and Raphidascarididae has varied according to both the genes analyzed and the types of analytical methods used (Nadler and Hudspeth, 1998; Zhu, Gasser, and Chilton, 1998; Nadler and Hudspeth, 2000; Shih, 2004)....
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TL;DR: Stomachs of 270 phocids from five species (bearded, Erignathus barbatus; hooded, Cystophora cristata; grey, Halichoerus grypus; harbour, Phoca vitulina; and ringed, P. hispida), collected off Newfoundland and Labrador during 1985-1993 were examined, and the numbers and percentages of larval and adult ascaridoid nematodes determined.
Abstract: Stomachs of 270 phocids from five species (bearded, Erignathus barbatus; hooded, Cystophora cristata; grey, Halichoerus grypus; harbour, Phoca vitulina; and ringed, P. hispida), collected off Newfo...
48 citations
"Molecular phylogenetics and diagnos..." refers background in this paper
...The most detailed studies of North American marine mammal ascaridoids have focused on hosts from Canadian Atlantic waters (Brattey and Ni, 1992; Brattey and Stenson, 1993; Brattey and Davidson, 1996)....
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TL;DR: The relationships between larvae and adults of Anisakis from the Mediterranean Sea and North-East Atlantic Ocean were analysed by multilocus electrophoresis and confirmed the correspondence of type I larvae with the A. simplex complex, including the sibling species A. physeteris is confirmed.
Abstract: The relationships between larvae and adults of Anisakis from the Mediterranean Sea and North-East Atlantic Ocean were analysed by multilocus electrophoresis. The correspondence of type I larvae with the A. simplex complex, including the sibling species A. simplex A and B, and of type II larvae with A. physeteris is confirmed. 19 of the 22 loci studied discriminated between the two larval types. Biochemical keys are given for the electrophoretic identification of A. simplex A, A. simplex B and A. physeteris, at both the larval and adult stages.
48 citations
"Molecular phylogenetics and diagnos..." refers background in this paper
...…allozyme methods, which have a long history of application to investigations of Anisakis diversity (Mattiucci et al., 1986; Nascetti et al., 1986; Orecchia et al., 1986; Paggi and Bullini, 1994; Bullini et al., 1997), have independently supported the validity of A. simplex s. str., A. typica, A.…...
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...…of genetically differentiated sibling species with different geographic and host distributions (Mattiucci et al., 1986; Nascetti et al., 1986; Orecchia et al., 1986; Paggi et al., 1991; Nascetti et al., 1993; Mattiucci et al., 1997, 1998; Paggi, Mattiucci et al., 1998; Paggi et al., 2000;…...
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TL;DR: Findings support the existence of two distinct sibling species, Contracaecum ogmorhini (sensu lato) and C. margolisi n.
Abstract: Genetic variation of Contracaecum ogmorhini (sensu lato) populations from different otariid seals of the northern and southern hemisphere was studied on the basis of 18 enzyme loci as well as preliminary sequence analysis of the mitochondrial cyt b gene (260 bp). Samples were collected from Zalophus californianus in the boreal region and from Arctocephalus pusillus pusillus, A. pusillus doriferus and A. australis from the austral region. Marked genetic heterogeneity was found between C. ogmorhini (sensu lato) samples from the boreal and austral region, respectively. Two loci (Mdh-2 and NADHdh) showed fixed differences and a further three loci (Iddh, Mdh-1 and 6Pgdh) were highly differentiated between boreal and austral samples. Their average genetic distance was DNei=0.36 at isozyme level. At mitochondrial DNA level, an average proportion of nucleotide substitution of 3.7% was observed. These findings support the existence of two distinct sibling species, for which the names C. ogmorhini (sensu stricto) and C. margolisi n. sp., respectively, for the austral and boreal taxon, are proposed. A description for C. margolisi n. sp. is provided. No diagnostic morphological characters have so far been detected; on the other hand, two enzyme loci, Mdh-2 and NADHdh, fully diagnostic between the two species, can be used for the routine identification of males, females and larval stages. Mirounga leonina was found to host C. ogmorhini (s.s.) in mixed infections with C. osculatum (s.l.) (of which C. ogmorhini (s.l.) was in the past considered to be a synonym) and C. miroungae; no hybrid genotypes were found, confirming the reproductive isolation of these three anisakid species. The hosts and geographical range so far recorded for C. margolisi n. sp. and C. ogmorhini (s.s.) are given.
46 citations
TL;DR: It is proposed that the PCR-RFLP assay described in this report may be used for identification of marine and freshwater parasites from superfamily Ascaridoidea.
46 citations
Additional excerpts
...…reaction (PCR) including PCR-restriction fragment length polymorphism (PCR-RFLP) (Zhu, Gasser, Podolska, and Chilton, 1998; D’Amelio et al., 2000; Kijewska et al., 2002; Shih, 2004), single-strand conformational polymorphism (SSCP) of PCR products (Zhu, Gasser, Podolska, and Chilton, 1998; Zhu…...
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