COI Haplotypes Research Articles

Genetic diversity of armored scales (Hemiptera: Diaspididae) and soft scales (Hemiptera: Coccidae) in Chile

Scale insects (Sternorrhyncha: Coccoidea) are one of the most invasive and agriculturally damaging insect groups. Their management and the development of new control methods are currently jeopardized by the scarcity of identification data, in particular in regions where no large survey coupling morphological and DNA analyses have been performed. In this study, we sampled 116 populations of armored scales (Hemiptera: Diaspididae) and 112 populations of soft scales (Hemiptera: Coccidae) in Chile, over a latitudinal gradient ranging from 18°S to 41°S, on fruit crops, ornamental plants and trees. We sequenced the COI and 28S genes in each population. In total, 19 Diaspididae species and 11 Coccidae species were identified morphologically. From the 63 COI haplotypes and the 54 28S haplotypes uncovered, and using several DNA data analysis methods (Automatic Barcode Gap Discovery, K2P distance, NJ trees), up to 36 genetic clusters were detected. Morphological and DNA data were congruent, except for three species (Aspidiotus nerii, Hemiberlesia rapax and Coccus hesperidum) in which DNA data revealed highly differentiated lineages. More than 50% of the haplotypes obtained had no high-scoring matches with any of the sequences in the GenBank database. This study provides 63 COI and 54 28S barcode sequences for the identification of Coccoidea from Chile.

Low genetic but high morphological variation over more than 1000\xa0km coastline refutes omnipresence of cryptic diversity in marine nematodes

BackgroundThe resilience of ecosystems to negative impacts is generally higher when high gene flow, species diversity and genetic diversity are present. Population genetic studies are suitable to investigate genetic diversity and estimate gene flow between populations. Seaweed beds form a dynamic shallow water ecosystem influenced by climate change and human exploitation, as such, seaweed beds are a particularly powerful model to investigate ecosystem resilience in coastal areas. We studied the population genetic structure of the new nematode species Paracanthonchus gynodiporata associated with seaweeds in northeastern Brazil. Nematodes are generally believed to have a limited dispersal capacity because of the lack of planktonic larvae. Yet, they can drift on seaweeds, and water currents might be a natural barrier for their dispersal. Populations of P. gynodiporata were sampled over more than 1000 km coastline in regions across major oceanic currents with and without historical exploitation of seaweed.ResultsP. gynodiporata is described in an integrative way using mitochondrial and nuclear sequences and morphological data. The 3D model of the head region shows for the first time a detailed view of the ventrosublateral teeth, a character often overlooked in older taxonomic studies of the genus. A total of 17 mitochondrial COI haplotypes were found with one haplotype representing 63 to 83% of the frequencies in each population. AMOVA showed overall little population genetic structure (FST = 0.05204), and no genetic subdivision between the populations under the influence of the two different water currents were found. Effects of historical seaweed exploitation on population genetic diversity were not detected. In contrast, significant differences between populations were found in morphometric characters. This discrepancy in genetic and morphological differentiation between populations across 1000 km of coastline is surprising in view of the frequently observed presence of several cryptic species at small geographical scale in other macroalgal associated nematodes.ConclusionsOur results show that cryptic species are not omnipresent in marine nematode species, suggesting that nematodes associated with seaweeds have been able to disperse over large distances across well-known biogeographic barriers.

Exotic aphid species Brachycaudus divaricatae in Central Europe: Distribution, host specificity and molecular diversity

AbstractAphid species

Geographic variation in the spotted-wing drosophila, Drosophila suzukii (Diptera: Drosophilidae), based on mitochondrial DNA sequences

The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), is an economically damaging pest that feeds on most thin-skinned fruits. It was originally native to a few Asian countries, including Korea, but is now found in North America and Europe. In this study, we sequenced portions of the mitochondrial (mt) COI and ND4 genes from a total of 195 individuals collected mainly from Korea. We then combined GenBank-registered COI sequences from all ancestral-range and introduced-range populations with our own COI data to assess the worldwide diversity, divergence, and relatedness of SWD haplotypes. A total of 139 haplotypes were obtained from the concatenated COI and ND4 sequences. Most haplotypes were confined to single localities, but 12 of them were found in more than two localities, and one haplotype (SWDCN61) was found from Korea to Canada. A dataset combining GenBank sequences with our own data identified a total of 94 worldwide COI haplotypes with a maximum sequence divergence (MSD) of 5.433% (32 bp). Although most haplotypes were found in only a single country, a few haplotypes were found commonly in China, Korea, and Japan; these occurred at a higher frequency and were often involved in introductions. A rough estimate of genetic diversity in each country showed higher diversity in ancestral distributional ranges, but the invasion over Asian countries seems to have been substantial because haplotype diversity was only 2.35 to 3.97-fold lower in the U.S.A, Canada, and Italy than that in the populations’ ancestral ranges.

Variation in the COI gene of the freshwater pearl mussel Margaritifera margaritifera from River Vuokkijoki

The freshwater pearl mussel Margaritifera margaritifera L. is one of the most endangered freshwater mussels in the world. Effective conservation of threatened species requires not only ecological, but also genetic information from the target species and populations. Since low genetic diversity can reduce the ability of a species to adapt to environmental changes, maintaining genetic diversity has been identified as one of the key elements in successful conservation programs. We examined genetic variation of the freshwater pearl mussel from the River Vuokkijoki, Karelia, Russia. We sequenced a fragment of the cytochrome c oxidase subunit I gene (COI) from 22 individuals and compared the data to 32 previously published COI sequences available in GenBank. We identified 10 different COI haplotypes in the sequenced samples, three of which had not been previously reported. Our results show that the River Vuokkijoki has high genetic diversity and suggest that the colonization of this northern freshwater pearl mussel population might have occurred from multiple and even distant refugia. Therefore, the freshwater pearl mussel population of the River Vuokkijoki is valuable for the conservation of the whole species.

Morphological and molecular characterisation of Heterodera koreana (Vovlas, Lamberti & Choo, 1992) Mundo-Ocampo, Troccoli, Subbotin, Del Cid, Baldwin & Inserra, 2008 (Nematoda: Heteroderidae) from bamboo in Japan

The Korean cyst nematode, Heterodera koreana, was recorded for the first time from Japan and characterised morphologically, morphometrically and molecularly. In total, 41 populations were detected from soil samples collected from the rhizosphere of four bamboo species in Japan: 31 populations from moso bamboo (Phyllostachys edulis), seven from madake (P. bambusoides), two from henon bamboo (P. nigra var. henonis) and one from fish pole bamboo (P. aurea). The morphology and morphometrics of the Japanese population were in agreement with those of the original description of H. koreana from South Korea and other subsequent descriptions from China and Iran, with the exception of some minor differences. The results of the phylogenetic analyses of the D2-D3 expansion segments of 28S rRNA gene and ITS rRNA gene sequences confirmed the species identification and phylogenetic relationship of H. koreana with other Heterodera species. The COI mtDNA gene sequences were obtained for the first time for H. koreana. Three COI haplotypes found in Japanese H. koreana populations showed a characteristic geographical distribution in Japan.

Seasonal Variability of the Paramecium aurelia Complex in the Botanical Garden of the Jagiellonian University, Krak6w - in the Light of Species Composition and COI Haplotype Variation.

The temporal occurrence of some Paramecium aurelia species is still an intriguing problem as cysts were never reported to exist in the Paramecium genus. A sequence of species occurrence was studied (by strain crosses and molecular identification) in five water-bodies of the Jagiellonian University Botanical Garden in Krak6w in different sampling sites and different seasons of the year. In the current study 20 P. aurelia strains were isolated from collected water samples and identified as P. biaurelia, P. tetraurelia, P. sexaurelia (the first record in Poland), P. novaurelia (the first record in the Botanical Garden). Generally only one species was found in the particular water body in a single sampling point in a given season - an exception was observed in the case of some strains of P. tetraurelia and P. sexaurelia. The latter species were mostly isolated from two water bodies situated in the Palm Houses (higher temperature preference) and P. biaurelia with P. novaurelia from water bodies located outside (lower temperature preference). Sequencing of the COImtDNA fragment revealed 9 haplotypes in the studied area which were characteristic for particular species. The most variable species was P. sexaurelia - 8 strains studied and 3 haplotypes identified. In contrast, P. novaurelia has only one haplotype for 6 strains collected in different seasons. The present study supports the hypothesis that botanical garden water bodies may be a hot-spot for microbial eukaryotic species-such as Paramecium.

New insights on geographical/ecological populations within Coilia nasus (Clupeiformes: Engraulidae) based on mitochondrial DNA and microsatellites

The Coilia nasus is an important fish species, which is of commercial values in China. In order to manage the fisheries resources and the development of sustainable fishing strategies to protect this species, 11 microsatellite loci polymorphisms and mtDNA COI sequences were used to examine the genetic diversity of C. nasus in Japan and China. In total, the 40 COI haplotypes and 93 microsatellite alleles were detected. The mtDNA phylogeny did not support population grouping, but the distribution patterns of mtDNA haplotypes and the results of STRUCTURE analysis based on microsatellite indicated a degree of genetic isolation in this species. Our study suggested that the lack of a population genetic structure might result in its amphidromous life cycle, and the geographical distance and habitat fragments might cause isolated populations. Thus, the sampling populations of C. nasus in Japan and China could be divided as four geographical/ecological populations.

Bless this phylogeographic mess – Comparative study of Eiseniella tetraedra (Annelida, Oligochaeta) between an Atlantic area and a continental Mediterranean area in Spain

Due to the influence of Atlantic Ocean and Mediterranean Sea, Spain has different climates, from desert to Atlantic. We sampled the parthenogenetic earthworm Eiseniella tetraedra in two different biogeographical zones in Spain, in order to study their genetic diversity and test their potential distinctiveness. Moreover, we evaluated the presence or absence of two different lineages (Eurosiberian and Mediterranean) found in other parthenogenetic earthworms such as Aporrectodea trapezoides and A. rosea. We studied the molecular markers COI, 16S and 28S. E. tetraedra presents a high diversity in Spain (one COI haplotype every two individuals were found) and no clear geographical patterns except for diffuse patterns along the Guadarrama River basin. In contrast, worldwide localities were more homogeneous with low diversity, to be confirmed with further samples. After morphological study, no correlation was found between phylogenetic relationships and the diagnostic characters for the previously described subspecies in E. tetraedra.

Phylogenetics, biogeography and population genetics of the ascidian Botryllus schlosseri in the Mediterranean Sea and beyond

The wide distribution of the ascidian Botryllus schlosseri along the Mediterranean coasts has been documented since the eighteenth century. However, despite copious documentation, analyses of dispersal modes and genetic profiles were limited to local populations or restricted regions. In order to get a pan-Mediterranean overview, 288 specimens from 11 populations of B. schlosseri from the western and eastern Mediterranean basins were sampled and analyzed using five microsatellite loci and COI sequences. Both molecular markers revealed high polymorphisms, with 182 microsatellites alleles and 54 COI haplotypes. Overall, Fst, Dest, and COI Фpt values were 0.146, 0.635 and 0.322, respectively, reflecting a high genetic diversity and a significant genetic structure as compared to other B. schlosseri populations worldwide, reflected by substantially higher values for effective number of alleles (Ne) in the Mediterranean. A phylogenetic analysis of the COI sequences resulted in four distinct clades and two molecular operational taxonomic units (OTUs). We recorded a stronger genetic structure among the populations of the eastern basin compared to the western basin (microsatellites Fst=0.217 versus 0.082; COI Фpt=0.416 versus 0.171), suggesting either a restricted connectivity between the basins or a stronger genetic drift in each basin. The occurrence of two OTUs and different ecological conditions may also contribute to this finding. Mean Nei’s genetic distance in the eastern Mediterranean populations was more than three times higher compared to the western basin. No correlation was observed between geographic and genetic distances (Mantel test), suggesting that maritime transport is the main dispersal vector of B. schlosseri colonies. The possibility that the Mediterranean is a center of diversity for B. schlosseri, and probably its site of origin, is further discussed.

Population Genetics of Overwintering Monarch Butterflies, Danaus plexippus (Linnaeus), from Central Mexico Inferred from Mitochondrial DNA and Microsatellite Markers.

Population genetic variation and demographic history in Danaus plexippus (L.), from Mexico were assessed based on analyses of mitochondrial cytochrome c oxidase subunit I (COI; 658 bp) and subunit II (COII; 503 bp) gene segments and 7 microsatellite loci. The sample of 133 individuals included both migratory monarchs, mainly from 4 overwintering sites within the Monarch Butterfly Biosphere Reserve (MBBR) in central Mexico (states of Michoacán and México), and a nonmigratory population from Irapuato, Guanajuato. Haplotype (h) and nucleotide (π) diversities were relatively low, averaging 0.466 and 0.00073, respectively, for COI, and 0.629 and 0.00245 for COII. Analysis of molecular variance of the COI data set, which included additional GenBank sequences from a nonmigratory Costa Rican population, showed significant population structure between Mexican migratory monarchs and nonmigratory monarchs from both Mexico and Costa Rica, suggesting limited gene flow between the 2 behaviorally distinct groups. Interestingly, while the COI haplotype frequencies of the nonmigratory populations differed from the migratory, they were similar to each other, despite the great physical distance between them. Microsatellite analyses, however, suggested a lack of structure between the 2 groups, possibly owing to the number of significant deviations from Hardy–Weinberg equilibrium resulting from heterzoygote deficiencies found for most of the loci. Estimates of demographic history of the combined migratory MBBR monarch population, based on the mismatch distribution and Bayesian skyline analyses of the concatenated COI and COII data set (n = 89) suggested a population expansion dating to the late Pleistocene (~35000–40000 years before present) followed by a stable effective female population size (Nef) of about 6 million over the last 10000 years.

Delimiting the species Neosiphonia yendoi (Rhodomelaceae, Rhodophyta) based on COI and rbcL genetic variation in Korea and Japan

Although the marine red algal genus Neosiphonia is well characterized, many species of Neosiphonia are poorly understood. To correctly define the species delimitation of Neosiphonia yendoi using genetic variation, owing to the confusion over identification with “N. sphaerocarpa” from Korea, we investigated intensively the haplotype network of the mitochondrial COI and the plastid rbcL genes of specimens collected from Korea and Japan. The molecular analyses indicated that specimens collected in different sites of Korea and Japan belong to the same species, Neosiphonia yendoi and “Neosiphonia sphaerocarpa” from Korea, which is distinguished from N. sphaerocarpa from Florida and is allied with N. yendoi collected from the type locality, Muroran of Japan. A total of 29 COI and 13 rbcL haplotypes were found and the COI haplotype network shows evidence of a clear break between specimens from Jeju Island and all other locations of Korea, suggesting the possibility of cryptic diversity within N. yendoi.

Erratum.

Previous articleNext article FreeErratum: The Biological Bulletin, Volume 228, no. 3, pp. 201–216Original articleEvidence of a Native Northwest Atlantic COI Haplotype Clade in the Cryptogenic Colonial Ascidian Botryllus schlosseriPDFPDF PLUSFull Text Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinked InRedditEmailQR Code SectionsMoreIn the article by Philip Yund, Catherine Collins, and Sheri Johnson, “Evidence of a native northwest Atlantic COI haplotype clade in the cryptogenic colonial ascinian Botryllus schlosseri” (June 2015; vol. 228, pp. 201–216), the new Bs38 haplotype, mentioned on p. 207, was detected in Falmouth, England, not Ireland. On p. 208, three haplotypes, sampled in Ireland, should be added to Table 3: HH (one copy), Bs18 (three copies), and Bs34 (four copies). The authors regret the errors. Previous articleNext article DetailsFiguresReferencesCited by The Biological Bulletin Volume 231, Number 1August 2016 Published in association with the Marine Biological Laboratory Article DOIhttps://doi.org/10.1086/689595 © 2016 by Marine Biological Laboratory. All rights reserved.PDF download Crossref reports no articles citing this article.Related articlesEvidence of a Native Northwest Atlantic COI Haplotype Clade in the Cryptogenic Colonial Ascidian Botryllus schlosseri26 Sep 2016The Biological Bulletin

Foul play? On the rapid spread of the brown shrimp Penaeus aztecus Ives, 1891 (Crustacea, Decapoda, Penaeidae) in the Mediterranean, with new records from the Gulf of Lion and the southern Levant

Specimens of the penaeid shrimp Penaeus aztecus, a West Atlantic species, were collected off Le Grau du Roi, Gulf of Lion, France, and off the Israeli coast, Levant Basin, Mediterranean Sea. This alien species has been previously recorded off Turkey, Greece, Montenegro and the Tyrrhenian coast of Italy. The species identity was confirmed based on morphological characters and by sequencing 406 nucleotides of the 16S RNA gene and 607 nucleotides of the COI. The 16S rRNA sequences of the specimens collected in Israel, France and Italy were identical, and exhibited three different COI haplotypes. The near-concurrent records from distant locations in the Mediterranean put paid to the premise that P. aztecus was introduced into the Mediterranean Sea in ballast waters. A more prudent proposition is that many of these populations issue from illegal introductions. Potential risks to native biodiversity and economic value are the likely competition with commercially important native prawns, co-introduction of pathogens and parasites, and risk of infecting penaeid populations elsewhere in the Mediterranean Sea with Erythraean alien disease agents previously limited to the Levant.

Preliminary evidence of differences in cadmium tolerance in metal-free stocks of the standard earthworm test species Eisenia andrei (Oligochaeta).

To test whether metal-tolerant and metal-sensitive earthworm specimens could be an inherent part of metal-free earthworm populations, (i) we used DNA barcoding to identify and categorize earthworms from 8 populations of the standard test species Eisenia andrei, and (ii) the earthworms carrying three of the identified COI haplotypes (named Hap1, hap3 and Hap3) were paired up and exposed to Cd in order to assess the difference in Cd sensitivity between the breeding pairs. A total of six breeding pairs were exposed to 0, 25, 50 and 100mg Cd/kg for 4weeks at 20°C. The survival of the breeding pairs, their change in biomass and cocoon production were assessed. For all of the endpoints assessed, the results indicated that couple 6 (Hap3×Hap3) was the most sensitive breeding pair whereas couple 4 (Hap1×Hap3) was the least sensitive one. The analysis of Cd tissue contents revealed that with increasing Cd concentration, Cp6 (Hap3×Hap3) could accumulate significantly more Cd than any other breeding pair (p≤0.01). Our findings indicate that E. andrei may harbour intrinsically Cd-tolerant and Cd-sensitive individuals and that this may be due to individual differences in Cd accumulation kinetics. In the context of ecotoxicological testing, our results underline the importance of using genetically diverse populations in laboratory testing to prevent generating flawed data from genetically homogeneous laboratory stocks. Although we do not regard the present mitochondrial haplotypes as proxy for possibly nuclear encoded traits, we discuss the necessity of a standardised earthworm barcoding protocol that could help not only to confirm the taxonomy of laboratory earthworm stocks but also to select genetically diverse stocks suitable for laboratory testing.

Species discovery and diversity in Lobocriconema (Criconematidae: Nematoda) and related plant-parasitic nematodes from North American ecoregions

There are many nematode species that, following formal description, are seldom mentioned again in the scientific literature. Lobocriconema thornei and L. incrassatum are two such species, described from North American forests, respectively 37 and 49 years ago. In the course of a 3-year nematode biodiversity survey of North American ecoregions, specimens resembling Lobocriconema species appeared in soil samples from both grassland and forested sites. Using a combination of molecular and morphological analyses, together with a set of species delimitation approaches, we have expanded the known range of these species, added to the species descriptions, and discovered a related group of species that form a monophyletic group with the two described species. In this study, 148 specimens potentially belonging to the genus Lobocriconema were isolated from soil, individually measured, digitally imaged, and DNA barcoded using a 721 bp region of cytochrome oxidase subunit 1 (COI). One-third of the specimens were also analyzed using amplified DNA from the 3' region of the small subunit ribosomal RNA gene (18SrDNA) and the adjacent first internal transcribed spacer (ITS1). Eighteen mitochondrial haplotype groups, falling into four major clades, were identified by well-supported nodes in Bayesian and maximum likelihood trees and recognized as distinct lineages by species delimitation metrics. Discriminant function analysis of a set of morphological characters indicated that the major clades in the dataset possessed a strong morphological signal that decreased in comparisons of haplotype groups within clades. Evidence of biogeographic and phylogeographic patterns was apparent in the dataset. COI haplotype diversity was high in the southern Appalachian Mountains and Gulf Coast states and lessened in northern temperate forests. Lobocriconema distribution suggests the existence of phylogeographic patterns associated with recolonization of formerly glaciated regions by eastern deciduous forest, but definitive glacial refugia for this group of plant parasitic nematodes have yet to be identified. Unlike agricultural pest species of plant-parasitic nematodes, there is little evidence of long-distance dispersal in Lobocriconema as revealed by haplotype distribution. Most haplotype groups were characterized by low levels of intragroup genetic variation and large genetic distances between haplotype groups. The localization of nematode haplotypes together with their characteristic plant communities could provide insight into the historical formation of these belowground biotic communities.

Limited dispersal in an ectoparasitic mite, Laelaps giganteus, contributes to significant phylogeographic congruence with the rodent host, Rhabdomys.

To explore how biogeography, parasite life history and host vagility influences evolutionary codivergences, we followed a comparative phylogeography approach using a host-specific nonpermanent mite, Laelaps giganteus, that occurs on four rodent species within the genus Rhabdomys. A mtDNA COI haplotype network derived for 278 parasite specimens showed marked phylogeographic congruence with host distributions. Analysis of the less variable nuclear intron Tropomyosin was in part consistent with these results. Although distance-based cophylogenetic analyses in axparafit failed to support significant mtDNA codivergences (P≥0.02), event-based analyses revealed significant cophylogeny between sampling localities of Rhabdomys and Laelaps using core-pa (P=0.046) and jane (P=0.026; P=0.00). These findings, in conjunction with the weak congruence previously reported among the permanent ectoparasitic lice Polyplax and Rhabdomys, suggest that host-parasite intimacy is not the most important driver of significant codivergence in our study system. Instead, the more restricted dispersal ability of L.giganteus, when compared to Polyplax, resulted in stronger spatial structuring and this could have resulted in significant codivergence. Host switching occurred predominantly on the edges of host distributions and was probably facilitated by climate-induced range shifts. When host ranges shift, the phylogeographic structure of L.giganteus is not reflecting the host movements as most of the nest bound parasites do not disperse with the host (they miss the boat) and the genetic contribution of the few dispersing mite individuals is often overwhelmed by the large number of individuals already present in nests within the new environment (causing them to drown on arrival).

Wolbachiaendosymbionts distort DNA barcoding in the parasitoid wasp genusDiplazon(Hymenoptera: Ichneumonidae)

Molecular species delimitation has the potential to speed up both discovery and description rates for new species. However, several studies have shown a limited performance of the standard DNA barcoding approach which relies on a single mitochondrial gene, cytochrome oxidase 1 (COI). Besides incomplete lineage sorting or a low substitution rate, distortion of mitochondrial inheritance patterns by bacterial endosymbionts could explain the failure of barcoding. Numerous reviews have discussed this phenomenon, but only few empirical examples exist. In the present study, we examine the effect of Wolbachia bacteria on barcoding in the parasitoid wasp genus Diplazon. Although integrative taxonomy recognizes 16 species, COI only recovers up to ten. Adding multivariate morphometrics, genotyping a fast-evolving nuclear gene (ITS2) and screening the Diplazon species for Wolbachia, we show that the failure of DNA barcoding coincides with the presence of the endosymbiont. Two species even share identical COI haplotypes and Wolbachia strains, even though ITS2 suggests that they are not closely related. This is one of very few examples of mitochondrial DNA introgression between well-established insect species, facilitated by Wolbachia. We review similar reports, provide a list of criteria to identify endosymbiont-mediated introgression, and discuss the prevalence and impact of this phenomenon on insect systematics and evolution.

Behavior and population structure of Anopheles darlingi in Colombia

Anopheles darlingi is a widely distributed and important malaria vector in Colombia. Biogeographical and ecological heterogeneity across the Colombian distribution led to the hypothesis of behavioral and genetic differentiation among A. darlingi populations. A total of 2017 A. darlingi specimens were collected during 222h of sampling. This vector was the most abundant anopheline species in most of the localities sampled. Subdivision between samples collected west and east of the Andes was indicated by 1) mitochondrial COI and nuclear CAD sequences from NW-W and CE-S populations (COI ΦST=0.48761–0.81974, CAD FST=0.11319–0.21321), 2) a COI haplotype network, and 3) SAMOVA. Endo- and exophagy were detected in populations west of the Andes, whereas exophagy was evident in PTG, a locality east of the Andes. Isolation by resistance was significant for COI and explained 26% of the genetic differentiation. We suggest that at a macrogeographic scale, the Andes influence the differentiation of A. darlingi in Colombia and may drive divergence, and, at a microgeographic scale, ecological differences have a significant impact on structure. These data could constitute a baseline for the design of effective vector interventions, locality-specific for the east and similar for panmictic populations west of the Andes.

Mitochondrial evidence supports a Nearctic origin for the spreading limicolous earthworm Sparganophilus tamesis Benham, 1892 (Clitellata, Sparganophilidae)

We analysed samples of Sparganophilus taken at the corners of its distribution area in Europe (UK, Germany and Italy). No mitochondrial genetic divergence within and amongst them was found, neither in COI nor in 16S. Further, the COI haplotype was also identical to two sequences from Ontario, Canada in the Barcoding of Life Data System (BOLD) database. Our European COI and 16S sequences showed only minimal differentiation (only 1 or 2 substitutions) from specimens newly collected in Illinois and Washington states (USA), as well as from a COI haplotype from Tennessee (USA) in BOLD. An additional COI haplotype from Illinois (found in BOLD) is 2.1% different from the other haplotypes but clearly belongs to the same lineage of Sparganophilus . This geographically broad but genetically compact group fits the morphological diagnosis of S. tamesis Benham, 1892 as revised by Jamieson (1971) and is seen as evidence that all European populations 1) belong to the same species, 2) derive from a recent introduction, 3) are conspecific with the most widespread species of Sparganophilus in North America, and that 4) S. tamesis is a senior synonym of S. eiseni Smith, 1895. The single European haplotype does not refute the possibility of its spread from a single introduced source population.