Putative hybrids between two Anisakis cryptic species: Molecular genotyping using High Resolution Melting

Cavallero, Serena; Costa, Antonella; Caracappa, Santo; Gambetta, Barbara; D’Amelio, Stefano

doi:10.1016/j.exppara.2014.08.017

Cited by 25 publications

(12 citation statements)

References 37 publications

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“…A. pegreffii or A. simplex (s. s.), when based on allozyme loci and the EF1 α −1 nDNA gene ( Table 2 ). On the other hand, the finding of numerous larval and adult hybrid specimens has previously been reported by several authors in those areas of the Mediterranean Sea (Farjallah et al 2008 ; Meloni et al 2011 ; Cavallero et al 2012 , 2014 ; Chaligiannis et al 2012 ; Pekmezci et al 2014 ; Molina-Fernández et al 2015 ), as just inferred from analyses of the ITS region of rDNA. In the present study, if the analysis of the ITS region of the rDNA gene had been the only conventional marker used in the identification of the parasites which we identified, a misidentification of these specimens of Anisakis would have resulted.…”

Section: Discussionmentioning

confidence: 61%

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No more time to stay ‘single’ in the detection ofAnisakis pegreffii, A. simplex(s. s.) and hybridization events between them: a multi-marker nuclear genotyping approach

et al. 2016

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SUMMARYA multi-marker nuclear genotyping approach was performed on larval and adult specimens of Anisakis spp. (N = 689) collected from fish and cetaceans in allopatric and sympatric areas of the two species Anisakis pegreffii and Anisakis simplex (s. s.), in order to: (1) identify specimens belonging to the parental taxa by using nuclear markers (allozymes loci) and sequence analysis of a new diagnostic nuclear DNA locus (i.e. partial sequence of the EF1 α−1 nDNA region) and (2) recognize hybrid categories. According to the Bayesian clustering algorithms, based on those markers, most of the individuals (N = 678) were identified as the parental species [i.e. A. pegreffii or A. simplex (s. s.)], whereas a smaller portion (N = 11) were recognized as F1 hybrids. Discordant results were obtained when using the polymerase chain reaction–restriction fragment length polymorphisms (PCR–RFLPs) of the internal transcribed spacer (ITS) ribosomal DNA (rDNA) on the same specimens, which indicated the occurrence of a large number of ‘hybrids’ both in sympatry and allopatry. These findings raise the question of possible misidentification of specimens belonging to the two parental Anisakis and their hybrid categories derived from the application of that single marker (i.e. PCR–RFLPs analysis of the ITS of rDNA). Finally, Bayesian clustering, using allozymes and EF1 α−1 nDNA markers, has demonstrated that hybridization between A. pegreffii and A. simplex (s. s.) is a contemporary phenomenon in sympatric areas, while no introgressive hybridization takes place between the two species.

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Section: Discussionmentioning

confidence: 61%

“… 2008 ; Meloni et al . 2011 ; Chaligiannis et al 2012 ; Cavallero et al 2012 , 2014 ; Pekmezci et al 2014 ) and sympatric areas (Abollo et al . 2003 ; Martìn-Sànchez et al .…”

Section: Introductionunclassified

No more time to stay ‘single’ in the detection ofAnisakis pegreffii, A. simplex(s. s.) and hybridization events between them: a multi-marker nuclear genotyping approach

et al. 2016

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“…in ascaridoid nematodes (Abollo et al, 2003;Peng et al, 2003;Cavallero et al, 2014). Cross-infection and hybridization between human and porcine species have been reported also in Trichuris spp.…”

Section: Discussionmentioning

confidence: 98%

Genetic heterogeneity and phylogeny of Trichuris spp. from captive non-human primates based on ribosomal DNA sequence data

Cavallero

Liberato

Friedrich

et al. 2015

Infection, Genetics and Evolution

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a b s t r a c tNematodes of the genus Trichuris, known as whipworms, are recognized to infect numerous mammalian species including humans and non-human primates. Several Trichuris spp. have been described and species designation/identification is traditionally based on host-affiliation, although cross-infection and hybridization events may complicate species boundaries. The main aims of the present study were to genetically characterize adult Trichuris specimens from captive Japanese macaques (Macaca fuscata) and grivets (Chlorocebus aethiops), using the ribosomal DNA (ITS) as molecular marker and to investigate the phylogeny and the extent of genetic variation also by comparison with data on isolates from other humans, non-human primates and other hosts. The phylogenetic analysis of Trichuris sequences from M. fuscata and C. aethiops provided evidences of distinct clades and subclades thus advocating the existence of additional separated taxa. Neighbor Joining and Bayesian trees suggest that specimens from M. fuscata may be distinct from, but related to Trichuris trichiura, while a close relationship is suggested between the subclade formed by the specimens from C. aethiops and the subclade formed by T. suis.The tendency to associate Trichuris sp. to host species can lead to misleading taxonomic interpretations (i.e. whipworms found in primates are identified as T. trichiura). The results here obtained confirm previous evidences suggesting the existence of Trichuris spp. other than T. trichiura infecting non-human living primates.

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“…Another primer-probe system was based on the ITS region of the rDNA, allowing the distinction between A. simplex (s. s.) and A. pegreffii (Mossali et al, 2010). Based on the same method, Cavallero et al (2014) proposed a HRM (high resolution melt) to identify two species (i.e. A. pegreffii and A. simplex (s. s.)) of the genus Anisakis.…”

Section: Introductionmentioning

confidence: 99%

Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis , Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)

et al. 2018

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Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in "seafood" products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.), P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum, a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency (E) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.

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Putative hybrids between two Anisakis cryptic species: Molecular genotyping using High Resolution Melting

Cited by 25 publications

References 37 publications

No more time to stay ‘single’ in the detection ofAnisakis pegreffii, A. simplex(s. s.) and hybridization events between them: a multi-marker nuclear genotyping approach

No more time to stay ‘single’ in the detection ofAnisakis pegreffii, A. simplex(s. s.) and hybridization events between them: a multi-marker nuclear genotyping approach

Genetic heterogeneity and phylogeny of Trichuris spp. from captive non-human primates based on ribosomal DNA sequence data

Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis , Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)

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