Molecular phylogeny of cuckoos supports a polyphyletic origin of brood parasitism

We constructed a molecular phylogeny of 15 species of cuckoos using mitochondrial DNA sequences spanning 553 nucleotide bases of the cytochrome b gene and 298 nucleotide bases of the ND2 gene. A parallel analysis for the cytochrome b gene including published sequences in the Genbank database was performed. Phylogenetic analyses of the sequences were done using parsimony, a sequence distance method (Fitch-Margoliash), and a character-state method which uses probabilities (maximum likelihood). Phenograms support the monophyly of three major clades: Cuculinae, Phaenicophaeinae and Neomorphinae-Crotophaginae. Clamator, a strictly parasitic genus traditionally included within the Cuculinae, groups together with Coccyzus (a nonobligate parasite) and some nesting cuckoos. Tapera and Dromococcyx, the parasitic cuckoos from the New World, appear as sister genera, close to New World cuckoos: Neomorphinae and Crotophaginae. Based on the results, and being conscious that a more strict resolution of the relationships among the three major clades is required, we postulate that brood parasitism has a polyphyletic origin in the Cuculiformes, with parasite species being found within the three defined clades. Evidence suggests that species within each clade share a common parasitic ancestor, but some show partial or total loss of brood parasitic behaviour.     

Monophyly and phylogeny of cuckoos (Aves, Cuculidae) inferred from osteological characters

A reanalysis of 32 characters from the literature previously deemed diagnostic of the Cuculidae revealed only five to be synapomorphic. I subsequently examined skeletons from 54 avian families and identified nine additional synapomorphies that supported cuckoo monophyly. My cladistic analysis of 33 cuculid genera using 135 skeletal characters differs markedly from currently accepted taxonomies. The most striking deviation is the placement of both New and Old World parasitic cuckoos in the Cuculinae, supporting the evolution of brood parasitism in a single event rather than three times as previously proposed. Unlike earlier classifications, the Cuculinae also includes the facultative parasites Coccyzus. This, suggests that the ancestral Coccyzus was an obligate parasite, and is consistent with the many behavioral adaptations to parasitism exhibited by this genus. Other changes include the placement of three subfamilies, comprising non-parasitic, terrestrial cuckoos of Old World (Centropodinae and Carpococcystinae) and New World (Neomorphinae) distribution, in basal positions on the tree. Nineteen characters support a sister relationship between the Hoatzin ( Opisthocamus hoatzin Müller) and turacos (Musophagidae), and not cuckoos. Three synapomorphies of the os carpi ulnare were found to unite the Cuculidae, turacos, and the Hoatzin, suggesting that these three diverse taxa may constitute a monophyletic group.     

Phylogenetic analysis of the New World Ptininae (Coleoptera: Bostrichoidea)

A phylogenetic analysis of the New World Ptininae (Anobiidae) was conducted with representatives of nine of ten New World genera, several Old World genera and seven more of the ten subfamilies of Anobiidae. One hundred and two characters (forty‐three multistate) from thirty‐four taxa were used. The single cladogram shows Ptininae as monophyletic and the sister group of the remaining Anobiidae, supporting their placement as subfamilies of a monophyletic Bostrichidae. Genus Niptus Boieldieu is polyphyletic supporting recognition of Pseudeurostus Heyden and the creation of a new genus to encompass the remaining New World species of Niptus. Flightlessness has evolved a minimum of three times within Ptininae and myrmecophily has probably evolved three times within just the New World taxa. The classifications of Ptininae and the remaining Anobiidae are examined and the evolution of feeding habits, myrmecophily and wing loss are discussed.     

Phylogeny and historical biogeography of the paper wasp genus Polistes (Hymenoptera: Vespidae): implications for the overwintering hypothesis of social evolution

The phylogeny of the paper wasp genus Polistes is investigated using morphological and behavioural characters, as well as molecular data from six genes (COI, 12S, 16S, 28S, H3, and EF1‐α). The results are used to investigate the following evolutionary hypotheses about the genus: (i) that Polistes first evolved in Southeast Asia, (ii) that dispersal to the New World occurred only once, and (iii) that long‐term monogyny evolved as an adaptation to overwintering in a temperate climate. Optimization of distribution records on the recovered tree does not allow unambiguous reconstruction of the ancestral area of Polistes. While the results indicate that Polistes dispersed into the New World from Asia, South America is recovered as the ancestral area for all New World Polistes: Nearctic species groups evolved multiple times from this South American stock. The final tree topology suggests strongly that the genus first arose in a tropical environment, refuting the idea of monogyny as an overwintering adaptation.     

New genus and new subgenus of subfamily Doryctinae (Hymenoptera: Braconidae) from the Old World fauna

New genus Doryctoproctus n. gen. (type species D. africanus n. sp.) from South Africa, and new subgenus of the genus Pedinotus Szépligeti (Eopedinotus n. subgen. (type species Pedinotus (Eopedinotus) inopinatus n. sp.) from Philippines are described. The genus Pedinotus is recorded in Old World for the first time.     

Three terrestrial Pleistocene coucals (Centropus: Cuculidae) from southern Australia: biogeographical and ecological significance

Coucals are large, predatory, primarily ground‐dwelling cuckoos of the genus Centropus, with 26 extant species ranging from Africa to Australia. Their evolutionary and biogeographical history are poorly understood and their fossil record almost non‐existent. Only one species (Centropus phasianinus) currently inhabits Australia, but there is now fossil evidence for at least three Pleistocene species. One of these (Centropus colossus) was described from south‐eastern Australia in 1985. Here we describe additional elements of this species from the same site, and remains of two further extinct species from the Thylacoleo Caves of the Nullarbor Plain, south‐central Australia. The skeletal morphology and large size of the three extinct species indicates that they had reduced capacity for flight and were probably primarily ground‐dwelling. The extinct species include the two largest‐known cuckoos, weighing upwards of 1 kg each. They demonstrate that gigantism in this lineage has been more marked in a continental context than on islands, contrary to the impression gained from extant species. The evolutionary relationships of the Australian fossil coucals are uncertain, but our phylogenetic analysis indicates a possible close relationship between one of the Nullarbor species and extant Centropus violaceus from the Bismarck Archipelago. The presence of three coucals in southern Australia markedly extends the geographical range of the genus from tropical Australia into southern temperate regions. This demonstrates the remarkable and consistent ability of coucals to colonize continents despite their very limited flying ability.     

A complete molecular phylogeny of Claravis confirms its paraphyly within small New World ground‐doves (Aves: Peristerinae) and implies multiple plumage state transitions

The three species in the genus Claravis (Aves: Peristerinae) are unique among members of the small New World ground‐dove clade. All three species inhabit forested areas rather than open scrubby habitat, and exhibit obvious sexual dichromatism. However, the phylogenetic relationships within Claravis remain unknown. The only molecular phylogenetic study to include more than one species of Claravis indicated the genus is paraphyletic. Here we include molecular data from all three Claravis species, including sequences from a museum skin of the previously unsampled Claravis geoffroyi (purple‐winged ground‐dove). Using both mitochondrial and nuclear loci, we estimate phylogenies and divergence times for the small New World ground‐dove clade. We also use ancestral state reconstruction methods to infer the evolution of male blue plumage (and thus sexual dimorphism) in the clade. As in the previous study we recover Claravis as a paraphyletic group, but with Claravis geoffroyi as the sister species to Claravis mondetoura (maroon‐chested ground‐dove). This result has important implications for the evolutionary history of the small New World ground‐dove clade. In particular, we recover multiple independent transitions between the monomorphic and dimorphic plumage states, which perhaps indicates sexual dimorphism arose twice in the group.     

Host‐specific parasitism in the Central American striped cuckoo,Tapera naevia

Coevolutionary theories of brood parasite strategy and host defense have been informed by research on egg mimicry and host recognition. However, there is no information on the strategies of New World parasitic cuckoos and their hosts. The striped cuckoo Tapera naevia is a New World cuckoo that uses multiple host species and maintains an egg color polymorphism. To investigate if color‐matching influenced rejection behavior in hosts, I conducted an egg rejection experiment on a host that lays blue‐green eggs, the rufous‐and‐white wren Thryophilus rufalbus and a host that lays white eggs, the plain wren Cantorchilus modestus. I used spectrophotometric analysis of egg color to determine the degree of egg color‐matching. I found that at the field site the striped cuckoo lays highly mimetic eggs for the rufous‐and‐white wren, in both color and brightness. The rufous‐and‐white wren was more likely to accept mimetic artificial eggs than non‐mimetic eggs. The plain wren exhibited low rejection rates for both mimetic and non‐mimetic artificial eggs. The evidence from this study indicates that the striped cuckoo lays eggs that are closely color‐matched to those of its preferred host, the rufous‐and‐white wren, and that this mimicry improves acceptance.     

Uncovering historical signature of mitochondrial DNA hidden in the nuclear genome: the biogeography of Schistocerca revisited

Inadvertent coamplification of nuclear mitochondrial pseudogenes (numts) is a serious problem in mitochondrial systematics, but numts can also be a valuable source of information because they represent ancient forms of mtDNA. We present a conceptual framework of numt accumulation, which states that in a given species there can be two types of numts, synaponumts and autaponumts, resulting from integration occurring respectively before and after a speciation event. In a given clade, a species that diverged early can only have its own autaponumts as well as synaponumts that were already present in the genome of the last common ancestor. A species that diverged more recently may, however, have many different synaponumts integrated at each different divergence as well as its own autaponumts. Therefore it is possible to decipher the evolutionary history of a species based on the phylogenetic distribution of numts in a simultaneous analysis of numts and extant mtDNA. In this study, we test this idea empirically in the context of addressing a controversial question regarding the biogeography of the grasshopper genus Schistocerca Stål (Orthoptera: Acrididae), based on numts of the cytochrome c oxidase subunit I (COI) gene. We find that our empirical data can be explained adequately by our conceptual framework, and that the phylogenetic distribution of COI numts reveals intricate evolutionary histories about past speciation events that are otherwise difficult to detect using conventional markers. Our study strongly favours the Old World origin of the desert locust, Schistocerca gregaria and the New World Schistocerca species are descendants from an ancestral gregaria‐like species that colonized the New World via westward transatlantic flight. However, the phylogenetic distribution of S. gregaria numts raises a distinct possibility that there might have been multiple founding events from Africa to America to give rise to the present‐day diversity of the genus. This is a case study for a creative use of numts as molecular fossils, and we demonstrate that numts provide an interesting and powerful phylogenetic signal, much more than what extant mtDNA or nuclear gene sequences might be able to provide.     

Phylogenetics and taxonomy of the New World leafy spurges,Euphorbia section Tithymalus (Euphorbiaceae)

The 480 species of leafy spurges, Euphorbia subgenus Esula, represent the main temperate radiation in the large genus Euphorbia. This group is distributed primarily in temperate Eurasia, but with smaller, disjunct centres of diversity in the mountains of the Old World tropics, in temperate southern Africa and in the New World. The majority of New World diversity (32 species) occurs in a single section, section Tithymalus. We analysed sequences of the nrITS and plastid ndhF, trnH‐psbA, trnS‐trnG and trnD‐trnT regions to reconstruct the phylogeny of section Tithymalus and to examine the origins and diversification of the species native to the New World. Our results indicate that the New World species of section Tithymalus form a clade that is sister to the widespread, weedy E. peplus. The New World species fall into two primary groups: a ‘northern annual clade’ from eastern North America and a diverse clade of both annual and perennial species that is divided into three subgroups. Within the second group, there is a small ‘southern annual clade’ from Texas and northern Mexico, a perennial ‘Brachycera clade’ from the western United States and northern Mexico, and a perennial ‘Esuliformis clade’ from montane areas of Mexico, Guatemala, Honduras and the Caribbean island of Hispaniola. Ancestral state reconstructions indicate that the annual habit probably evolved in the ancestor of E. peplus and the New World clade, with a subsequent reversal to the perennial habit. In conjunction with this phylogenetic framework, the New World species of section Tithymalus are comprehensively reviewed. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 191–228.     

Evolution and phylogeny of the New Zealand cicada genus Kikihia Dugdale (Homoptera: Auchenorrhyncha: Cicadidae) with special reference to the origin of the Kermadec and Norfolk Islands' species

Aim Determine the phylogeny and dispersal patterns of the cicada genus Kikihia in New Zealand and the origin of the Norfolk, Kermadec, and Chatham Island cicadas. Location New Zealand, Norfolk Island, Kermadec Islands and Chatham Island. Methods DNA sequences from 16 species and four soon to be described species of cicadas from New Zealand and Norfolk Island (Australia) were examined. A total of 1401 base pairs were analysed from whole genome extraction of three mitochondrial genes (cytochrome oxidase subunit II, ATPase6 and ATPase8). These DNA sequences were aligned and analysed using standard likelihood approaches to phylogenetic analysis. Dates of divergences between clades were determined using a molecular clock based on Bayesian statistics. Results Most species in the genus Kikihia diverged between 3 and 5 million years ago (Ma) coincident with a period of rapid mountain building in New Zealand. Cicada species on the Kermadec and Norfolk Islands invaded recently from New Zealand and are closely related to the New Zealand North Island species Kikihia cutora. Main conclusions Speciation in the genus Kikihia was likely due in large part to the appearance of new habitats associated with the rise of the Southern Alps, starting c. 5 Ma. Dispersal of Kikihia species within mainland New Zealand probably occurred gradually rather than through long‐distance jumps. However, invasion of Norfolk, the Kermadecs and Chatham Islands had to have occurred through long‐distance dispersal.     

The concept of genus within the family Phytoseiidae (Acari: Parasitiformes): historical review and phylogenetic analyses of the genus Neoseiulus Hughes

Systematic studies on the family Phytoseiidae were first conducted at the beginning of the 20^th century but increased greatly after the Second World War. Various classifications have been proposed based on different characters such as: dorsal, ventral, and leg chaetotaxy; the shape of ventrianal and sternal shields; the shape of the insemination apparatus (spermatheca) and spermatodactylus; the number of teeth on the movable digit of chelicera; and dorsal and ventral adenotaxy. The genus concepts developed over the last five decades can be divided into two main categories or hypotheses. The first, supported mainly by Chant and McMurtry, focuses on dorsal and ventral chaetotaxy, and the genera so defined usually include a great number of species. The second category, proposed by Athias‐Henriot, considers the shape of the insemination apparatus as the key character, and the genera so defined usually include a limited number of species. From a diagnostic point of view, both classifications have a valid structure, but the question investigated herein was: which of the two classifications or hypotheses fits phylogenetic evolution? To answer this, we conducted molecular phylogenetic analyses (using the genes ITS and 12S rRNA) on the genus Neoseiulus, which has been subjected to classification based on the two main genus concepts. The results showed that the first hypothesis (Chant and McMurtry) leads to polyphyly of the genus Neoseiulus, while the second (Athias‐Henriot) leads to paraphyly of the genus. The results show that acarologists who first decided that the insemination apparatus was of evolutive importance could be correct as the shape of the insemination apparatus seems to better fit evolutive clades than dorsal and ventral chaetotaxy. The morphology of this organ, however, must be more accurately studied to better define homologies. The present paper investigates the two main hypotheses proposed until now for classification of Phytoseiidae and thereby opens the way for improved classification. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 165 , 253–273.     

Tropical Asian species show that the Old World clade of ‘spiny solanums’ (Solanum subgenus Leptostemonum pro parte: Solanaceae) is not monophyletic

The tropical Asian taxa of the species‐rich genus Solanum (Solanaceae) have been less well studied than their highly diverse New World relatives. Most of these tropical Asian species, including the cultivated brinjal eggplant/aubergine and its wild progenitor, are part of the largest monophyletic Solanum lineage, the ‘spiny solanums’ (subgenus Leptostemonum or the Leptostemonum clade). Here we present the first phylogenetic analysis of spiny solanums that includes broad sampling of the tropical Asian species, with 42 of the 56 currently recognized species represented. Two nuclear and three plastid regions [internal transcribed spacer (ITS), waxy, ndhF‐rpL32, trnS‐trnG and trnT‐trnF] were amplified and used to reconstruct phylogenetic relationships using maximum likelihood and Bayesian methods. Our analyses show that Old World spiny solanums do not resolve in a single clade, but are part of three unrelated lineages, suggesting at least three independent introductions from the New World. We identify and describe several monophyletic groups in Old World solanums that have not been previously recognized. Some of these lineages are coherent in terms of morphology and geography, whereas others show considerable morphological variation and enigmatic distribution patterns. Tropical Asia occupies a key position in the biogeography of Old World spiny solanums, with tropical Asian taxa resolved as the closest relatives of diverse groups of species from Australia and Africa.     

Multiple nuclear genes stabilize the phylogenetic backbone of the genus Quercus

Phylogenetic relationships among 108 oak species (genus Quercus L.) were inferred using DNA sequences of six nuclear genes selected from the existing genomic resources of the genus. Previous phylogenetic reconstructions based on traditional molecular markers are inconclusive at the deeper nodes. Overall, weak phylogenetic signals were obtained for each individual gene analysis, but stronger signals were obtained when gene sequences were concatenated. Our data support the recognition of six major intrageneric groups Cyclobalanopsis, Cerris, Ilex, Quercus, Lobatae and Protobalanus. Our analyses provide resolution at deeper nodes but with moderate support and a more robust infrageneric classification within the two major clades, the ‘Old World Oaks’ (Cyclobalanopsis, Cerris, Ilex) and ‘New World Oaks’ (Quercus, Lobatae, Protobalanus). However, depending on outgroup choice, our analysis yielded two alternative placements of the Cyclobalanopsis clade within the genus Quercus. When Castanea Mill. was chosen as outgroup, our data suggested that the genus Quercus comprised two clades corresponding to two subgenera as traditionally recognized by Camus: subgenus Euquercus Hickel and Camus and subgenus Cyclobalanopsis Øersted (Schneider). However, when Notholithocarpus Manos, Cannon and S. Oh was chosen as an outgroup subgenus Cyclobalanopsis clustered with Cerris and Ilex groups to form the Old World clade. To assess the placement of the root, we complemented our dataset with published data of ITS and CRC sequences. Based on the concatenated eight gene sequences, the most likely root position is at the split between the ‘Old World Oaks’ and the ‘New World Oaks’, which is one of the alternative positions suggested by our six gene analysis. Using a dating approach, we inferred an Eocene age for the primary divergences in Quercus and a root age of about 50–55 Ma, which agrees with palaeobotanical evidence. Finally, irrespective of the outgroup choice, our data boost the topology within the New World clade, where (Protobalanus + Quercus) is a sister clade of Lobatae. Inferred divergence ages within this clade and the Cerris–Ilex clade are generally younger than could be expected from the fossil record, indicating that morphological differentiation pre-dates genetic isolation in this clade.     

Intra‐specific variability in Rhinoleucophenga punctulata populations (Diptera: Drosophilidae) from Neotropical biomes: Combined analyses of morphological and molecular data

Despite the fact that Drosophilidae is a very diverse and well‐studied taxon, the New World genus Rhinoleucophenga is yet poorly understood even in regard to species distribution and morphological variability pattern. In this sense, R. punctulata is a species widely distributed in the Neotropical region. Specimens of R. punctulata were collected from different biomes in Brazil: Pampa, Cerrado and Caatinga sensu strictu, and a southern Amazonian savannah enclave area. Geographical variations in the external body morphology and in the morphology of spermatheca were noticed among the different populations. The hypothesis that each population could be a different species was tested through molecular data. A fragment of the mitochondrial cytochrome c oxydase subunit I (COI) gene was sequenced to perform phylogenetic analyses through neighbor‐joining and Bayesian inferences. Pairwise genetic divergences of COI sequences were calculated using DNA barcode premises. The analyzed populations presented different variation levels in both morphology and molecular traits. However, new species were not proposed because the intra‐population nucleotide variations exceeded the inter‐population ones. The noticeable morphological and genetic variations revealed among the four studied populations of R. punctulata in different biomes of Brazil suggest the necessity that morphological, distributional and molecular data at the population level should be integrated into complementary datasets to better understand the biological diversity of Rhinoleucophenga through Neotropical environments.     

Extraordinary Predation by the Neotropical Army Ant Cheliomyrmex andicola: Implications for the Evolution of the Army Ant Syndrome1

Workers of the genus Cheliomyrmex are unique among the New world army ants (subfamily Ecitoninae) in that their mandibles are armed with elongate, spine‐like teeth. We present the first prey records for this genus. Cheliomyrmex andicola prey on large‐bodied ground dwelling invertebrates and, possibly, on vertebrates. Unlike other army ants, C. andicola workers use their sting during prey capture. The workers' unusual mandibles and potent stings may be adapted for piercing and gripping the integument of nonarthropod prey animals, and for rapidly subduing large‐bodied prey, respectively. The genus Cheliomyrmex may be the sister taxon to other Neotropical army ants (Ecitoninae), and Cheliomyrmex shares features of mandibular morphology and prey selection with Old World driver ants in the genus Dorylus. Mass cooperative foraging, an important element of army ant behavior, may have arisen in part as an adaptation for exploiting large‐bodied prey.     

Phylogeny of the tribe Empoascini (Hemiptera: Cicadellidae: Typhlocybinae) based on morphological characteristics,with reclassification of the Empoasca generic group

A morphology‐based phylogenetic analysis of the tribe Empoascini (Hemiptera: Cicadellidae: Typhlocybinae) is presented for 58 of 83 formerly recognized genera based on 99 morphological characters of adults. The results support excluding the New World Beamerana generic group from Empoascini. The remaining genera of Empoascini were recovered as a monophyletic sister group of Dikraneurini. Previously recognized tribes Jorumini and Helionini are derived from within Empoascini and are considered synonyms of the latter tribe. Three previously recognized informal generic groups, the Empoasca group, Alebroides group and Usharia group were paraphyletic but the Ficiana group was recovered as monophyletic based on five synapomorphies. Genera previously placed in the Alebroides group represent at least six independent lineages, indicating that the hind wing character separating this group from the Empoasca group (CuA and MP veins free) is highly homoplasious. Empoasca (sensu lato) is also paraphyletic. Thus, twelve previously recognized subgenera of Empoasca are elevated to genus status and five species groups of Empoasca from the New World are recognized as separate new genera. Sikkimasca Dworakowska, 1993 is treated as synonym of Marolda Dworakowska, 1977 based on the phylogeny. Biogeographic analysis suggests that Empoascini most likely first evolved in the Oriental region and spread to other biogeographic realms more recently by multiple independent invasions.     

CHROMOSOME NUMBERS IN COMPOSITAE. VI. SENECIONEAE. II

Chromosome counts are reported for 76 taxa and 2 natural hybrids of tribe Senecioneae (Compositae). First counts are reported for several species of Senecioneae as well as for the genera Cadiscus and Whitneya. New chromosome numbers are added to those previously known in Arnica, Cacalia, and Senecio. Additional counts from Arnica support our previous suggestion that x = 19 for this genus. It is assumed that observed meiotic irregularities are associated with apomixis in this genus. Basic chromosome numbers for various New World sections of Senecio are proposed, and certain problems of sectional relationships in this genus are discussed. Chromosome numbers and plant morphology of Cadiscus, Hulsea, and Whitneya indicate that these genera should be removed from Helenieae to Senecioneae. The possible affinity of the anomalous genus Adenocaulon with Mutisieae is discussed. Data presented in the paper further support our earlier proposal that the basic chromosome number for Senecioneae is x = 10.     

At least two indigenous species of the Bemisia tabaci complex are present in Brazil

Bemisia tabaci is one of the most important global agricultural insect pests, being a vector of emerging plant viruses such as begomoviruses and criniviruses that cause serious problems in many countries. Although knowledge of the genetic diversity of B. tabaci populations is important for controlling this pest and understanding viral epidemics, limited information is available on this pest in Brazil. A survey was conducted in different locations of São Paulo and Mato Grosso states, and the phylogenetic relationships of B. tabaci individuals from 43 populations sampled from different hosts were analysed based on partial mitochondrial cytochrome oxidase 1 gene (mtCOI) sequences. According to the recently proposed classification of the B. tabaci complex, which employs the 3.5% mtCOI sequence divergence threshold for species demarcation, most of the specimens collected were found to belong to the Middle East‐Asia Minor 1 species, which includes the invasive populations of the commonly known B biotype, within the Africa/Middle East/Asia Minor high‐level group. Three specimens collected from Solanun gilo and Ipomoea sp. were grouped together and could be classified in the New World species that includes the commonly known A biotype. However, six specimens collected from Euphorbia heterophylla, Xanthium cavanillesii and Glycine maxima could not be classified into any of the 28 previously proposed species, although according to the 11% mtCOI sequence divergence threshold, they belong to the New World high‐level group. These specimens were classified into a new recently proposed species named New World 2 that includes populations from Argentina. Middle East‐Asia Minor 1, New World and New World 2 were differentiated by RFLP analysis of the mtCOI gene using TaqI enzyme. Taq I analysis in silico also differentiates these from Mediterranean species, thus making this method a convenient tool to determine population dynamics, especially critical for monitoring the presence of this exotic pest in Brazil.     

A large‐scale molecular survey of Clinostomum (Digenea,Clinostomidae)

Members of the genus Clinostomum Leidy, 1856 are parasites that mature in birds, with occasional reports in humans. Because morphological characters for reliable discrimination of species are lacking, the number of species considered valid has varied by an order of magnitude. In this study, sequences from the DNA barcode region of cytochrome c oxidase I (CO1) and/or internal transcribed spacer (ITS) from specimens from Mexico, Bolivia, Peru, Brazil, Kenya, China and Thailand were analysed together with published sequences from Europe, Africa, Indonesia and North America. Although ITS and CO1 distances among specimens were strongly correlated, distance‐based analysis of each marker yielded different groups. Putative species indicated by CO1 distances were consistent with available morphological identifications, while those indicated by ITS conflicted with morphological identifications in three cases. There was little overlap in sequence variation within and between species, particularly for CO1. Although ITS and CO1 distances tended to increase in specimens that were further apart geographically, this did not impair distance‐based species delineation. Phylogenetic analysis suggests a deep division between clades of Clinostomum inhabiting the New World and Old World, which parallels the distribution of their principal definitive hosts, the Ardeidae.