The parasite species complex Anisakis simplex sensu lato (Anisakis simplex sensu stricto; (A. simplex s.s.), A. pegreffii, A. simplex C) is the main cause of severe anisakiasis (allergy) worldwide and is now an important health matter. In this study, the relationship of this Anisakis species complex and their allergenic capacities is assessed by studying the differences between the two most frequent species (A. simplex s.s., A. pegreffii) and their hybrid haplotype by studying active L3 larvae parasiting Merluccius merluccius. They were compared by 2D gel electrophoresis and parallel Western blot (2DE gels were hybridized with pools of sera from Anisakis allergenic patients). Unambiguous spot differences were detected and protein assignation was made by MALDI-TOF/TOF analysis or de novo sequencing. Seventy-five gel spots were detected and the corresponding proteins were identified. Differentially expressed proteins for A. simplex s.s., A. pegreffii, and their hybrid are described and results are statistically supported. Twenty-eight different allergenic proteins are classified according to different families belonging to different biological functions. These proteins are described for the first time as antigenic and potentially new allergens in Anisakis. Comparative proteomic analyses of allergenic capacities are useful for diagnosis, epidemiological surveys, and clinical research. All MS data have been deposited in the ProteomeXchange with identifier PXD000662 (http://proteomecentral.proteomexchange.org/dataset/PXD000662).

BackgroundAnisakis simplex sensu stricto and Anisakis pegreffii are sibling species of nematodes parasitic on marine mammals. Zoonotic human infection with third stage infective larvae causes anisakiasis, a debilitating and potentially fatal disease. These 2 species show evidence of hybridisation in geographical areas where they are sympatric. How the species and their hybrids differ is still poorly understood.ResultsThird stage larvae of Anisakis simplex s.s., Anisakis pegreffii and hybrids were sampled from Merluccius merluccius (Teleosti) hosts captured in waters of the FAO 27 geographical area. Specimens of each species and hybrids were distinguished with a diagnostic genetic marker (ITS). RNA was extracted from pools of 10 individuals of each taxon. Transcriptomes were generated using Illumina RNA-Seq, and assembled de novo. A joint assembly (here called merged transcriptome) of all 3 samples was also generated. The inferred transcript sets were functionally annotated and compared globally and also on subsets of secreted proteins and putative allergen families. While intermediary metabolism appeared to be typical for nematodes in the 3 evaluated taxa, their transcriptomes present strong levels of differential expression and enrichment, mainly of transcripts related to metabolic pathways and gene ontologies associated to energy metabolism and other pathways, with significant presence of excreted/secreted proteins, most of them allergens. The allergome of the 2 species and their hybrids has also been thoroughly studied; at least 74 different allergen families were identified in the transcriptomes.ConclusionsA. simplex s.s., A. pegreffi and their hybrids differ in gene expression patterns in the L3 stage. Strong parent-of-origin effects were observed: A. pegreffi alleles dominate in the expression patterns of hybrids albeit the latter, and A. pegreffii also display significant differences indicating that hybrids are intermediate biological entities among their parental species, and thus of outstanding interest in the study of speciation in nematodes. Analyses of differential expression based on genes coding for secreted proteins suggests that co-infections presents different repertoires of released protein to the host environment. Both species and their hybrids, share more allergen genes than previously thought and are likely to induce overlapping disease responses.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4970-9) contains supplementary material, which is available to authorized users.

Forficula auricularia (the European earwig) is possibly a complex of cryptic species. To test this hypothesis, we performed: (1) a phylogeographic study based on fragments of the mitochondrial COI and the nuclear ITS2 markers on a wide geographic sampling, (2) morphometric analyses of lineages present in Spain and (3) niche overlap analyses. We recovered five reciprocally monophyletic ancient phylogroups with unique historical patterns of distribution, climatic niches and diversification. External morphology was conserved and not correlated with speciation events, except in one case. Phylogenetic placement of the morphologically distinct taxon renders F. auricularia paraphyletic. Based on the congruence of the phylogenetic units defined by mtDNA and nuclear sequence data, we conclude that phylogroups have their own historical and future evolutionary trajectory and represent independent taxonomic units. Forficula auricularia is a complex of at least four species: the morphologically diagnosable Forficula aeolica González-Miguéns & García-París sp. nov., and the cryptic taxa: Forficula mediterranea González-Miguéns & García-París sp. nov., Forficula dentataFabricius, 1775stat. nov. and Forficula auriculariaLinnaeus, 1758s.s. We also provide new synonymy for F. dentata.

Flightless, phoretic insects of the tribe Meloini include several widespread species able to inhabit both mainland and oceanic and continental islands. Relationships between phoretic and non‐phoretic species of Meloini are unclear, precluding broad evolutionary and biogeographic analyses within the group. We provide a robust molecular phylogenetic framework for Meloini, with finer sampling in the Western Palaearctic, by analysing two nuclear (Wg, 18S) and two mtDNA (cox1, 16S) gene fragments of 24 species representing eight subgenera of Meloe and Physomeloe. Our phylogenetic analyses revealed that Meloe, as currently recognized, is paraphyletic. Our phylogenetic hypothesis clarifies several controversial relationships within the group. We provide a new taxonomic framework that better reflects the evolutionary relationships of the group by raising to genus all previously considered subgenera: Afromeloe Schmidt, 1913, Alveomeloe Pripisnova, 1987, Chiromeloe Reitter, 1911, Desertimeloe Kaszab, 1964, Eurymeloe Reitter, 1911, Lampromeloe Reitter, 1911, Lasiomeloe Reitter, 1911, Listromeloe Reitter, 1911, Meloegonius Reitter, 1911, Mesomeloe Reitter, 1911, Meloenellus Reitter, 1911, Micromeloe Reitter, 1911, Taphromeloe Reitter, 1911 and Treiodous Dugès, 1869; while Anchomeloe Iablokoff‐Khnzorian, 1983, is recovered as subgenus of Meloe. In addition, we conducted an extensive intraspecific sampling for four phoretic taxa (Meloe proscarabaeus, Meloe autumnalis, Eurymeloe mediterraneus and Eurymeloe tuccia) and used mtDNA phylogeographic analyses to unveil patterns of overseas dispersal. Our phylogeographic analyses revealed a high level of geographically unstructured haplotype diversity within taxa, suggesting that transmarine dispersal events have occurred multiple times along the evolutionary history of phoretic species of blister beetles. Larval phoresy may explain the existence of idiosyncratic biogeographic and phylogeographic patterns in these flightless organisms.

Exoskeletons of beetles and their associated morphological characteristics can serve many different functions, including thermoregulation. We study the thermal role of the exoskeleton in 13 Geotrupidae dung beetle species using heating experiments under controlled conditions. The main purpose was to measure the influence of heating sources (solar radiance vs. infrared), animal position (dorsal exposure vs. ventral exposure), species identity, and phylogenetic relationships on internal asymptotic temperatures and heating rates. The thermal response was significantly influenced by phylogenetic relatedness, although it was not affected by the apterous condition. The asymptotic internal temperature of specimens was not affected by the thoracic volume but was significantly higher under simulated sunlight conditions than under infrared radiation and when exposed dorsally as opposed to ventrally. There was thus a significant interaction between heating source and body position. Heating rate was negatively and significantly influenced by thoracic volume, and, although insignificantly slower under simulated sunlight, it was significantly affected by body position, being faster under dorsal exposure. The results constitute the first evidence supporting the hypothesis that the beetle exoskeleton acts differentially across the electromagnetic spectrum determining internal body temperatures. This interesting finding suggests the existence of a kind of passive physiology imposed by the exoskeleton and body size, where interspecific relationships play a minor role.