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      Cross-species utility of microsatellite loci for the genetic characterisation of Anisakis berlandi (Nematoda: Anisakidae) Translated title: Utilité des loci microsatellites pour la caractérisation génétique interspécifique d’ Anisakis berlandi (Nematoda, Anisakidae)

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          Eight microsatellite loci, recently developed in the species Anisakis pegreffii, were successfully amplified in Anisakis berlandi, sibling species of the A. simplex (s. l.) complex. They were validated on adult specimens ( n = 46) of the parasite species, collected from two individuals of the definitive host, the long-finned pilot whale Globicephala melas from New Zealand waters. Among the eight loci scored, one, Anisl 07132, had null alleles in A. berlandi and was thus excluded from the subsequent genetic analysis. Two loci, Anisl 00314 and Anisl 10535, were monomorphic. In addition, as also previously detected in the other species of the A. simplex (s. l.) complex, the Anisl 7 locus was seen to be sex-linked, showing hemizygosity in male specimens. Differential allele frequency distributions of A. berlandi, with respect to those previously observed in A. pegreffii and A. simplex (s. s.), were found at some microsatellite loci. The Anisl 7 locus provided 100% diagnosis between A. berlandi and A. pegreffii, while others resulted in 99% diagnosis between A. berlandi and the other two species. Simple sequence repeat (SSR) loci also allowed us to estimate the genetic differentiation of A. berlandi from A. pegreffii ( F st ≈ 0.45, Dc = 0.82) and A. simplex (s. s.) ( F st ≈ 0.57, Dc = 0.73). The results suggest that SSRs provide a set of candidate markers for population genetics analysis of A. berlandi, as well as for the investigation, through a multi-locus genotyping approach, of possible patterns of hybridisation/introgression events between A. berlandi and the other two Anisakis species in sympatric conditions.

          Translated abstract

          Huit loci microsatellites, récemment développés chez l’espèce Anisakis pegreffii, ont été amplifiés avec succès chez Anisakis berlandi, espèce sœur du complexe A. simplex (s. l.). Ils ont été validés sur des spécimens adultes ( n = 46) de l’espèce, récoltés chez deux individus de l’hôte définitif, le globicéphale commun Globicephala melas, des eaux néo-zélandaises. Parmi les huit loci notés, l’un, Anisl 07132, avait des allèles nuls chez A. berlandi et a donc été exclu de l’analyse génétique ultérieure. Deux loci, Anisl 00314 et Anisl 10535, étaient monomorphes. De plus, comme cela a également été détecté précédemment dans les autres espèces du complexe A. simplex (s. l.), le locus Anisl 7 était lié au sexe, montrant une hémizygosité chez les spécimens mâles. Chez A. berlandi, des distributions de fréquences d’allèles, différentielles par rapport à celles précédemment observées chez A. pegreffii et A. simplex (s. s.), ont été trouvées pour certains loci microsatellites. Le locus Anisl 7 a fourni un diagnostic à 100 % entre A. berlandi et A. pegreffii, tandis que d’autres ont abouti à un diagnostic à 99 % entre A. berlandi et les deux autres espèces. Les loci des SSR ont également permis d’estimer la différenciation génétique d’ A. berlandi par rapport à A. pegreffii ( F st ≈ 0,45, Dc = 0,82) et A. simplex (s. s.) ( F st ≈ 0,57, Dc = 0,73). Les résultats suggèrent que les répétitions de séquences simples (SSR) fournissent un ensemble de marqueurs candidats pour l’analyse génétique des populations d’ A. berlandi, ainsi que pour l’investigation, dans une approche de génotypage multilocus, des modèles possibles d’hybridation/introgression entre A. berlandi et les deux autres espèces d’ Anisakis dans des conditions sympatriques.

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          Most cited references 26

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          The magnitudes of the systematic biases involved in sample heterozygosity and sample genetic distances are evaluated, and formulae for obtaining unbiased estimates of average heterozygosity and genetic distance are developed. It is also shown that the number of individuals to be used for estimating average heterozygosity can be very small if a large number of loci are studied and the average heterozygosity is low. The number of individuals to be used for estimating genetic distance can also be very small if the genetic distance is large and the average heterozygosity of the two species compared is low.
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              Advances and trends in the molecular systematics of anisakid nematodes, with implications for their evolutionary ecology and host-parasite co-evolutionary processes.

              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.

                Author and article information

                EDP Sciences
                11 February 2020
                : 27
                : ( publisher-idID: parasite/2020/01 )
                [1 ] Department of Public Health and Infectious Diseases, Section of Parasitology, Sapienza-University of Rome P.le Aldo Moro 5 00185 10 Rome Italy
                [2 ] Department of Ecological and Biological Sciences, Tuscia University Viale dell’Università s/n 01100 Viterbo Italy
                [3 ] Cawthron Institute 98 Halifax Street East, The Wood 7010 Nelson New Zealand
                Author notes

                Laboratory Affiliated to Istituto Pasteur Italia-Fondazione Cenci-Bolognetti, Viale Regina Elena, 291, 00161 Roma, Italy

                [* ]Corresponding author: simonetta.mattiucci@ 123456uniroma1.it
                parasite190119 10.1051/parasite/2020004
                © E. Bello et al., published by EDP Sciences, 2020

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 3, Tables: 3, Equations: 0, References: 25, Pages: 9
                Research Article


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