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Phylogenetic position of Phthiraptera (Insecta: Paraneoptera) and elevated rate of evolution in mitochondrial 12S and 16S rDNA.

Phthiraptera (chewing and sucking lice) and Psocoptera (booklice and barklice) are closely related to each other and compose the monophyletic taxon Psocodea. However, there are two hypotheses regarding their phylogenetic relationship: (1) monophyletic Psocoptera is the sister group of Phthiraptera or (2) Psocoptera is paraphyletic, and Liposcelididae of Psocoptera is the sister group of Phthiraptera. Each hypothesis is supported morphologically and/or embryologically, and this problem has not yet been resolved. In the present study, the phylogenetic position of Phthiraptera was examined using mitochondrial 12S and 16S rDNA sequences, with three methods of phylogenetic analysis. Results of all analyses strongly supported the close relationship between Phthiraptera and Liposcelididae. Results of the present analyses also provided some insight into the elevated rate of evolution in mitochondrial DNA (mtDNA) in Phthiraptera. An elevated substitution rate of mtDNA appears to originate in the common ancestor of Phthiraptera and Liposcelididae, and directly corresponds to an increased G+C content. Therefore, the elevated substitution rate of mtDNA in Phthiraptera and Liposcelididae appears to be directional. A high diversity of 12S rDNA secondary structure was also observed in wide range of Phthiraptera and Liposcelididae, but these structures seem to have evolved independently in different clades.

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Multiple origins of parasitism in lice: phylogenetic analysis of SSU rDNA indicates that the Phthiraptera and Psocoptera are not monophyletic.

The Paraneoptera (Hemipteroid Assemblage) comprises the orders Thysanoptera (thrips), Hemiptera (bugs), Phthiraptera (lice) and Psocoptera (booklice and barklice). The phylogenetic relationships among the Psocodea (Phthiraptera and Psocoptera), Thysanoptera and Hemiptera are unresolved, as are some relationships within the Psocodea. Here, we present phylogenetic hypotheses inferred from SSU rDNA sequences; the most controversial of which is the apparent paraphyly of the Phthiraptera, which are parasites of birds and mammals, with respect to one family of Psocoptera, the Liposcelididae. The order Psocoptera and the suborder that contains the Liposcelididae, the Troctomorpha, are also paraphyletic. The two remaining psocopteran suborders, the Psocomorpha and the Trogiomorpha, are apparently monophyletic. The Liposcelididae is most closely related to lice from the suborder Amblycera. These results suggest that the taxonomy of the Psocodea needs revision. In addition, there are implications for the evolution of parasitism in insects; parasitism may have evolved twice in lice or have evolved once and been subsequently lost in the Liposcelididae.

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Phylogenetic analysis of partial sequences of elongation factor 1alpha identifies major groups of lice (Insecta: Phthiraptera).

As a first attempt to use molecular data to resolve the relationships between the four suborders of lice and within the suborder Ischnocera, we sequenced a 347-bp fragment of the elongation factor 1alpha gene of 127 lice (Insecta: Phthiraptera) as well as outgroup taxa from the order Psocoptera. A number of well-supported monophyletic groups were found but the relationships among many of these groups could not be resolved. While it is probable that multiple substitutions at high divergences and ancient radiation over a short period of time have contributed to the problem, we attribute most of this lack of resolution to the high ratio of taxa to characters. Nevertheless, the sequence data unequivocally support a number of important relationships that are at variance with the conclusions of morphological taxonomy. These include the sister group relationship of Chelopistes and Oxylipeurus, two lice occupying different ecological niches on the same host, which have previously been assigned to different families. These results provide evidence in support of the hypothesis that lice have speciated in situ on the host in response to niche specialization and that this has given rise to convergent morphologies in the lice of different host groups which share similar ecological niches. We discuss our attempts to overcome the limitations of this large data set, including the use of leaf stability analysis, a new method for analyzing the stability of taxa in a phylogenetic tree, and examine a number of hypotheses of relationships based on both traditional taxonomy and host associations.

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Multiple genes and the monophyly of Ischnocera (Insecta: Phthiraptera).

Whereas most traditional classifications identify Ischnocera as a major suborder of lice in the order Phthiraptera, a recent molecular study based on one gene did not recover monophyly of Ischnocera. In this study we test the monophyly of Ischnocera using sequences of portions of three different genes: two nuclear (EF1 alpha and 18S) and one mitochondrial (COI). Analysis of EF1 alpha and COI sequences did not recover monophyly of Ischnocera, but these genes provided little support for ischnoceran paraphyly because homoplasy is high among the divergent taxa included in this study. Analysis of 18S sequences recovered ischnoceran monophyly with strong support. Sequences from these three gene regions showed significant conflict with the partition homogeneity test, but this heterogeneity probably arises from the dramatic differences in substitution rates. In support of this conclusion, Kishino-Hasegawa tests of the EF1 alpha and COI genes did not reject several trees containing ischnoceran monophyly. Combined analysis of all three gene regions supported monophyly of Ischnocera, although not as strongly as analysis of 18S by itself. In sum, although rapidly evolving genes can retain some phylogenetic signal for deep phylogenetic relationships, strong support for such relationships is likely to come from more slowly evolving genes.

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Differences in straggling rates between two genera of dove lice (Insecta: Phthiraptera) reinforce population genetic and cophylogenetic patterns.

Differences in dispersal abilities have been implicated for causing disparate evolutionary patterns between Columbicola and Physconelloides lice (Insecta: Phthiraptera). However, no study has documented straggling (when lice are found on atypical hosts) rates within these lineages. We used the fact that the Galapagos Hawk, Buteo galapagoensis (Gould) (Falconiformes) feeds on the Galapagos Dove Zenaida galapagoensis Gould (Columbiformes) within an ecologically simplified setting. The Galapagos Dove is the only typical host of Columbicola macrourae (Wilson) and Physconelloides galapagensis (Kellogg and Huwana) in Galapagos. We quantitatively sampled and found these lice on both bird species. A DNA barcoding approach confirmed that stragglers were derived from Galapagos doves. We also collected a Bovicola sp. louse, likely originating from a goat (Capra hircus). On hawks, C. macrourae was significantly more prevalent than P. galapagensis. On doves, the two lice were equally prevalent and abundant. Differences in prevalence on hawks was a function of differences in straggling rate between lice, and not a reflection of their relative representation within the dove population. This provides further evidence that differences in dispersal abilities may drive differences in the degree of cospeciation in Columbicola and Phyconelloides lice, which have become model systems in evolutionary biology.

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Dramatically elevated rate of mitochondrial substitution in lice (Insecta: Phthiraptera).

Few estimates of relative substitution rates, and the underlying mutation rates, exist between mitochondrial and nuclear genes in insects. Previous estimates for insects indicate a 2-9 times faster substitution rate in mitochondrial genes relative to nuclear genes. Here we use novel methods for estimating relative rates of substitution, which incorporate multiple substitutions, and apply these methods to a group of insects (lice, Order: Phthiraptera). First, we use a modification of copath analysis (branch length regression) to construct independent comparisons of rates, consisting of each branch in a phylogenetic tree. The branch length comparisons use maximum likelihood models to correct for multiple substitution. In addition, we estimate codon-specific rates under maximum likelihood for the different genes and compare these values. Estimates of the relative synonymous substitution rates between a mitochondrial (COI) and nuclear (EF-1alpha) gene in lice indicate a relative rate of several 100 to 1. This rapid relative mitochondrial rate (>100 times) is at least an order of magnitude faster than previous estimates for any group of organisms. Comparisons using the same methods for another group of insects (aphids) reveals that this extreme relative rate estimate is not simply attributable to the methods we used, because estimates from aphids are substantially lower. Taxon sampling affects the relative rate estimate, with comparisons involving more closely related taxa resulting in a higher estimate. Relative rate estimates also increase with model complexity, indicating that methods accounting for more multiple substitution estimate higher relative rates.

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The population genetics of host specificity: genetic differentiation in dove lice (Insecta: Phthiraptera).

Some species of parasites occur on a wide range of hosts while others are restricted to one or a few host species. The host specificity of a parasite species is determined, in part, by its ability to disperse between host species. Dispersal limitations can be studied by exploring the genetic structure of parasite populations both within a single species of host and across multiple host species. In this study we examined the genetic structure in the mitochondrial cytochrome oxidase I (COI) gene of two genera of lice (Insecta: Phthiraptera) occurring on multiple sympatric species of doves in southern North and Central America. One genus, Columbicola, is generally less host-specific than the other, Physconelloides. For both genera we identified substantial genetic differentiation between populations of conspecific lice on different host species, generally 10-20% sequence divergence. This level of divergence is in the range of that often observed between species of these two genera. We used nested clade analysis to explore fine scale genetic structure within species of these feather lice. We found that species of Physconelloides exhibited more genetic structure, both among hosts and among geographical localities, than did species of Columbicola. In many cases, single haplotypes within species of Columbicola are distributed on multiple host species. Thus, the population genetic structure of species of Physconelloides reveals evidence of geographical differentiation on top of high host species specificity. Underlying differences in dispersal biology probably explain the differences in population genetic structure that we observed between Columbicola and Physconelloides.

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Damalinia (Tricholipeurus) zaganseeri, n. sp. (Phthiraptera: Trichodectidae), a parasite from Procapra gutturosa (Artiodactyla: Bovidae) in Mongolia.

During an expedition through the east Mongolian steppes, an undescribed chewing louse (Insecta: Phthiraptera) of the genus Damalinia was collected on Mongolian gazelles, Procapra gutturosa. This represents the 1st report of a chewing louse on this poorly known antelope. After a short review of the distribution of this genus and its subgenera among host groups, the new species D. (Tricholipeurus) zaganseeri, n. sp. is described and illustrated. Its distinct morphology is compared with that of similar species.

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Extraordinary number of gene rearrangements in the mitochondrial genomes of lice (Phthiraptera: Insecta).

The arrangement of genes in the mitochondrial (mt) genomes of most insects is the same, or near-identical, to that inferred to be ancestral for insects. We sequenced the entire mt genome of the small pigeon louse, Campanulotes bidentatus compar, and part of the mt genomes of nine other species of lice. These species were from six families and the three main suborders of the order Phthiraptera. There was no variation in gene arrangement among species within a family but there was much variation in gene arrangement among the three suborders of lice. There has been an extraordinary number of gene rearrangements in the mitochondrial genomes of lice!

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Structures of the preantennal region of several species of Damalinia (Phthiraptera: Trichodectidae).

A comparative study was made of the structures of the preantennal region of nine species of Damalinia Mjöberg, 1910 (Phthiraptera: Trichodectidae). Variability in the form of the head, ventral carina, pulvinus, mandibles, and labrum was observed by light and scanning electron microscopy. According to previous studies, the form of the head is possibly influenced by the density and texture of the hair of the host. The ventral carina, which maybe sharp or blunt, and the pulvinus, a membranous lobe, of these parasite species of different hosts, were studied. Photographs of the mandibles and labrum show a high variability according to species.

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Rallicola deckeri n. sp. (Phthiraptera: Philopteridae) from Ruddy Woodcreepers Dendrocincla homochroa (Passeriformes: Furnariidae) collected in Campeche, Mexico.

A new species of Rallicola Johnston and Harrison (Phthiraptera: Philopteridae) is described from lice collected from a series of Ruddy Woodcreepers Dendrocincla homochroa (Aves: Dendrocolaptinae) in Campeche, Mexico. Rallicola deckeri is easily recognized by the unique shape of the mesosome in the male and by the pattern of divided sternites on the female.

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Molecular systematics of Goniodidae (Insecta: Phthiraptera).

The higher level phylogenetic relationships within the avian feather lice (Insecta: Phthiraptera: Ischnocera) are extremely problematic. Here we investigate the relationships of 1 family (Goniodidae), sometimes recognized as distinct within Ischnocera, using parsimony and likelihood analyses of nuclear and mitochondrial DNA sequences. These data support monophyly for a restricted definition of traditional Goniodidae, but recognition of this family would result in paraphyly of the large heterogeneous family Philopteridae. We show that the New World Chelopistes is not related to other members of Goniodidae, despite similarities in morphology, but rather is the sister taxon to Oxylipeurus. Within Goniodidae, genera are divided into those occurring on Galliformes (the Goniodes complex) and those occurring on Columbiformes (the Coloceras complex). Within the well-sampled Coloceras complex, or Physconelloidinae, several groups are identified. However, traditionally recognized genera such as Coloceras and Phvsconelloides appear to be paraphyletic. Whereas the phylogeny of Goniodidae reflects some aspects of host relationships, biogeography also influences coevolutionary history.

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Critical evaluation of five methods for quantifying chewing lice (Insecta: Phthiraptera).

Five methods for estimating the abundance of chewing lice (Insecta: Phthiraptera) were tested. To evaluate the methods, feral pigeons (Columba livia) and 2 species of ischnoceran lice were used. The fraction of lice removed by each method was compared, and least squares linear regression was used to determine how well each method predicted total abundance. Total abundance was assessed in most cases using KOH dissolution. The 2 methods involving dead birds (body washing and post-mortem-ruffling) provided better results than 3 methods involving live birds (dust-ruffling, fumigation chambers, and visual examination). Body washing removed the largest fraction of lice (>82%) and was an extremely accurate predictor of total abundance (r2 = 0.99). Post-mortem-ruffling was also an accurate predictor of total abundance (r2 > or = 0.88), even though it removed a smaller proportion of lice (<70%) than body washing. Dust-ruffling and fumigation chambers removed even fewer lice, but were still reasonably accurate predictors of total abundance, except in the case of data sets restricted to birds with relatively few lice. Visual examination, the only method not requiring that lice be removed from the host, was an accurate predictor of louse abundance, except in the case of wing lice on lightly parasitized birds.

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Nervous system of poultry lice, Lipeurus lawrensis tropicalis Peters (Phthiraptera: Ischnocera).

The anatomy and histology of nervous system of Lipeurus lawrensis tropicalis (Phthiraptera: Ischnocera) is described in detail. The brain is U-shaped compact unit. It is tilted backwards to cover the tritocerebral commissure, circumoesophageal conncetives and a large portion of sub-oesophageal ganglion. The ventral nerve cord consists of sub-oesophageal ganglion and three thoracic ganglia. The abdominal ganglia are wanting and the abdomen is innervated by the nerves arising from the metathoracic ganglionic mass. Interganglionic connective are absent between the thoracic ganglia, but the sub-oesophageal ganglion and the prothoracic ganglion are connected by interganglionic connectives. The origin and disposition of different nerves arising from metathoracic ganglion show that it is a compound ganglionic mass, in which abdominal ganglia are also fused. From this ganglionic mass three pairs of nerves are given in thoracic segments and three pairs to the abdominal segments. The frontal ganglion is situated anterior to the brain at a considerable distance. The histology of the brain and that of thoracic ganglion is also described.

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Abrocomaphthirus chilensis (Gomez), new combination (Phthiraptera-Anoplura), an ectoparasite of the Chilean rodent Abrocoma bennetti (Abrocomidae).

The same species of sucking louse (Phthiraptera-Anoplura) from the Chilean abrocomid rodent Abrocoma bennetti Waterhouse, 1837 was recently described as Eulinognathus chilensis Gomez, 1998 and as Abrocomaphthirus hoplai Durden & Webb, 1999. We discuss the nomenclature of this louse and confirm that its correct name is now Abrocomaphthirus chilensis (Gomez, 1998). After comparing type specimens designated from each description, we also document some morphological variation within this species with respect to the paratergal plate on abdominal segment 3 in the female which usually has two relatively long apical setae, but occasionally has only one.

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Sequencing of a new target genome: the Pediculus humanus humanus (Phthiraptera: Pediculidae) genome project.

The human body louse, Pediculus humanus humanus (L.), and the human head louse, Pediculus humanus capitis, belong to the hemimetabolous order Phthiraptera. The body louse is the primary vector that transmits the bacterial agents of louse-borne relapsing fever, trench fever, and epidemic typhus. The genomes of the bacterial causative agents of several of these aforementioned diseases have been sequenced. Thus, determining the body louse genome will enhance studies of host-vector-pathogen interactions. Although not important as a major disease vector, head lice are of major social concern. Resistance to traditional pesticides used to control head and body lice have developed. It is imperative that new molecular targets be discovered for the development of novel compounds to control these insects. No complete genome sequence exists for a hemimetabolous insect species primarily because hemimetabolous insects often have large (2000 Mb) to very large (up to 16,300 Mb) genomes. Fortuitously, we determined that the human body louse has one of the smallest genome sizes known in insects, suggesting it may be a suitable choice as a minimal hemimetabolous genome in which many genes have been eliminated during its adaptation to human parasitism. Because many louse species infest birds and mammals, the body louse genome-sequencing project will facilitate studies of their comparative genomics. A 6-8X coverage of the body louse genome, plus sequenced expressed sequence tags, should provide the entomological, evolutionary biology, medical, and public health communities with useful genetic information.

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Hoplopleura janzeni n. sp. (Phthiraptera: Anoplura), a new sucking louse from a Central American swimming mouse.

Both sexes of a new species of sucking louse Hoplopleura janzeni (Phthiraptera: Hoplopleuridae) are described and illustrated from the Central American ichthyomyine swimming mouse Rheomys raptor (Rodentia: Muridae) collected in Costa Rica. The morphology of the new species is compared with that of Hoplopleura exima Johnson, the only other species of sucking louse known to parasitize an ichthyomyine rodent. Hoplopleura janzeni is unique in having posteriorly directed spurs on the first antennal segment, the fore- and midcoxae, and the hind femora of both sexes.

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