Molecular phylogenetics of the avian feather louse Philopterus-complex (Phthiraptera: Philopteridae).

The avian feather louse Philopterus-complex (Phthiraptera: Ischnocera: Philopteridae) currently contains 12 genera that have been grouped together because of shared morphological characteristics. Although previously lumped into a single genus (Philopterus), more recent morphological treatments have separated the group into several different genera. Here we evaluate the status of these genera using DNA sequence data from 118 ingroup specimens belonging to ten genera in the Philopterus-complex: Australophilopterus Mey, 2004, Cinclosomicola Mey 2004, Clayiella Eichler, 1940, Corcorides Mey, 2004, Mayriphilopterus Mey, 2004, Paraphilopterus Mey 2004, Philopteroides Mey 2004, Philopterus Nitzsch, 1818, Tyranniphilopterus Mey, 2004, and Vinceopterus Gustafsson, Lei, Chu, Zou, and Bush, 2019. Our sampling includes 97 new louse-host association records. Our analyses suggest that the genus Debeauxoecus Conci, 1941, parasitic on pittas (Aves: Pittidae), is outside of the Philopterus-complex, and that there is strong support for the monophyly of a group containing the remaining genera from the complex. Some diverse genera, such as Philopterus (sensu stricto) and Mayriphilopterus are supported as monophyletic, whereas the genera Australophilopterus, Philopteroides, and Tyranniphilopterus are not. The present study is the largest phylogenetic reconstruction of avian lice belonging to the Philopterus-complex to date and suggests that further generic revision is needed in the group to integrate molecular and morphological information.

Pichia bovicola sp. nov., a yeast species associated with small-intestinal content of cattle.

Three yeast strains, DMKU-MP6-4T, DMKU-MP2-6 and DMKU-MP5-1, were isolated from the small-intestinal content or Pia of cattle in Thailand during the investigation of yeast diversity in this habitat. According to the D1/D2 domain of the large subunit (LSU) rRNA gene and internal transcribed spacer (ITS) region sequence analysis, these strains represent a novel yeast species in the genus Pichia. The species produced one to four ascospores per ascus with spherical to ellipsoidal shape and heterogenous in terms of size. These three strains were identical and differed from their closely related species, Pichia exigua NRRL Y-10920T by 2% (six nucleotide substitutions and five gaps) in the D1/D2 domain of the large subunit rRNA gene, while ITS region sequences differed by 3.1% (16 nucleotide substitutions and 27 gaps), 3.7% (19 nucleotide substitutions and 28 gaps) and 3.1% (16 nucleotide substitutions and 27 gaps) for DMKU-MP6-4T, DMKU-MP2-6 and DMKU-MP5-1, respectively. The name, Pichia bovicola, is proposed to accommodate these species. The holotype is DMKU-MP6-4T (TBRC 15616T=PYCC 8905T).

Combining Nuclear and Mitochondrial Loci Provides Phylogenetic Information in the Philopterus Complex of Lice (Psocodea: Ischnocera: Philopteridae).

The Philopterus Complex includes several lineages of lice that occur on birds. The complex includes the genera Philopterus (Nitzsch, 1818; Psocodea: Philopteridae), Philopteroides (Mey, 2004; Psocodea: Philopteridae), and many other lineages that have sometimes been regarded as separate genera. Only a few studies have investigated the phylogeny of this complex, all of which are based on morphological data. Here we evaluate the utility of nuclear and mitochondrial loci for recovering the phylogeny within this group. We obtained phylogenetic trees from 39 samples of the Philopterus Complex (Psocodea: Philopteridae), using sequences of two nuclear (hyp and TMEDE6) and one mitochondrial (COI) marker. We evaluated trees derived from these genes individually as well as from concatenated sequences. All trees show 20 clearly demarcated taxa (i.e., putative species) divided into five well-supported clades. Percent sequence divergence between putative species (~5-30%) for the COI gene tended to be much higher than those for the nuclear genes (~1-15%), as expected. In cases where species are described, the lineages identified based on molecular divergence correspond to morphologically defined species. In some cases, species that are host generalists exhibit additional underlying genetic variation and such cases need to be explored by further future taxonomic revisions of the Philopterus Complex.

Integrative taxonomic description of the chewing louse Tricholipeurus lipeuroides infesting Odocoileus virginianus veraecrucis white-tailed deer in Veracruz, Mexico.

Chewing lice (Phthiraptera: Amblycera, Ischnocera) represent a component of the ectoparasite fauna associated with large sized mammals as deers. However, the diversity of chewing louse species infesting deer remains to be fully characterized in the Neotropics. Little is known about the chewing lice infesting the extant fourteen subspecies of white-tailed deer (Odocoileus virginianus) in Mexico. Known to infest white-tailed deer (WTD) in Canada and the United States (U.S.), Tricholipeurus lipeuroides is a chewing louse species that was originally described in the nineteenth century infesting O. v. mexicanus in Mexico. For the first time, infestation of O. v. veraecrucis, a Neotropical WTD subspecies in Mexico, with T. lipeuroides is reported herein. An integrative taxonomic approach was taken by combining morphological and molecular analyses to describe the T. lipeuroides infestion of O. v. veraecrucis. Ecological parameters of the T. lipeuroides infestations were also calculated. The prevalence was 91.7% of the 56 O. v. veraecrucis (29 females and 27 males) inspected while under chemical restraint that were sampled at 3 sites in the central region of Veracruz state in Mexico. The amplification and sequencing of previously reported T. lipeuroides Cytochrome Oxidase Subunit I gene confirmed the identity of all the chewing louse life stages. These results are discussed in the context of comparative analyses on the emergence of novel chewing lice-deer associations.

Rapid detection of Bovicola ovis using colourimetric loop-mediated isothermal amplification (LAMP): a potential tool for the detection of sheep lice infestation on farm.

The sheep body louse, Bovicola ovis (B. ovis), is one of the most significant ectoparasites affecting Australia's sheep flocks. Despite this, detection methods for B. ovis infestation are limited to visual inspection and ELISA. A colourimetric loop-mediated isothermal amplification (LAMP) method was developed and evaluated for the detection of B. ovis DNA. Diagnostic sensitivity and specificity of LAMP were compared with those of visual inspection and PCR and validated using field samples collected from 22 farms. Two different DNA extraction methods using a commercial kit and a boiling method were also compared. The highest sensitivity and specificity were observed when PCR was used and DNA was extracted using a commercial kit. Compared with PCR, the LAMP assay demonstrated a sensitivity and specificity of 90% and 92% when DNA was extracted by a commercial kit and 100% and 75% when DNA was extracted by the boiling method, respectively. The LAMP test developed in this study could potentially serve as a point-of-care diagnostic tool for monitoring of sheep flocks as well as surveillance of B. ovis populations.

Loss of genetic diversity, recovery and allele surfing in a colonizing parasite, Geomydoecus aurei.

Understanding the genetic consequences of changes in species distributions has wide-ranging implications for predicting future outcomes of climate change, for protecting threatened or endangered populations and for understanding the history that has led to current genetic patterns within species. Herein, we examine the genetic consequences of range expansion over a 25-year period in a parasite (Geomydoecus aurei) that is in the process of expanding its geographic range via invasion of a novel host. By sampling the genetics of 1,935 G. aurei lice taken from 64 host individuals collected over this time period using 12 microsatellite markers, we test hypotheses concerning linear spatial expansion, genetic recovery time and allele surfing. We find evidence of decreasing allelic richness (AR) with increasing distance from the source population, supporting a linear, stepping stone model of spatial expansion that emphasizes the effects of repeated bottleneck events during colonization. We provide evidence of post-bottleneck genetic recovery, with average AR of infrapopulations increasing about 30% over the 225-generation span of time observed directly in this study. Our estimates of recovery rate suggest, however, that recovery has plateaued and that this population may not reach genetic diversity levels of the source population without further immigration from the source population. Finally, we employ a grid-based sampling scheme in the region of ongoing population expansion and provide empirical evidence for the power of allele surfing to impart genetic structure on a population, even under conditions of selective neutrality and in a place that lacks strong barriers to gene flow.

The chewing lice (Phthiraptera: Ischnocera, Amblycera) of the great cormorant (Phalacrocorax carbo).

The Great Cormorant is a widespread bird species with almost worldwide distribution. Accordingly, its general biology has been investigated thoroughly. Less well known, however, are the chewing lice that live inside the plumage of this diving bird. We examined the two known species of Great Cormorant chewing lice, Eidmanniella pellucida (Rudow, 1869) (Amblycera: Menoponidae) and Pectinopygus gyricornis (Denny, 1842) (Ischnocera: Philopteridae). Taking advantage of the autofluorescence of the cuticle, confocal laser scanning microscopy (CLSM) was used to explore the external morphology of all developmental stages of P. gyricornis. Morphometric analyses revealed a standard increase in body size from first larval instar to the adult. In addition, all instars exhibited increasing body segment differentiation, especially in the abdomen and the head. A total of 277 individuals of Pectinopygus gyricornis and 2 individuals of Eidmanniella pellucida were collected from eleven Great Cormorants from Mecklenburg-Western Pomerania, Germany, in 2015.

Development and Characterization of 12 Novel Polymorphic Microsatellite Loci for the Mammal Chewing Louse Geomydoecus aurei (Insecta: Phthiraptera) and a Comparison of Next-Generation Sequencing Approaches for Use in Parasitology.

Next-generation sequencing methodologies open the door for evolutionary studies of wildlife parasites. We used 2 next-generation sequencing approaches to discover microsatellite loci in the pocket gopher chewing louse Geomydoecus aurei for use in population genetic studies. In one approach, we sequenced a library enriched for microsatellite loci; in the other approach, we mined microsatellites from genomic sequences. Following microsatellite discovery, promising loci were tested for amplification and polymorphism in 390 louse individuals from 13 pocket gopher hosts. In total, 12 loci were selected for analysis (6 from each methodology), none of which exhibited evidence of null alleles or heterozygote deficiencies. These 12 loci showed adequate genetic diversity for population-level analyses, with 3-9 alleles per locus with an average HE per locus ranging from 0.32 to 0.70. Analysis of Molecular Variance (AMOVA) indicated that genetic variation among infrapopulations accounts for a low, but significant, percentage of the overall genetic variation, and individual louse infrapopulations showed FST values that were significantly different from zero in the majority of pairwise infrapopulation comparisons, despite all 13 infrapopulations being taken from the same locality. Therefore, these 12 polymorphic markers will be useful at the infrapopulation and population levels for future studies involving G. aurei. This study shows that next-generation sequencing methodologies can successfully be used to efficiently obtain data for a variety of evolutionary questions.

The Mitochondrial Cytochrome Oxidase Subunit I Gene Occurs on a Minichromosome with Extensive Heteroplasmy in Two Species of Chewing Lice, Geomydoecus aurei and Thomomydoecus minor.

In animals, mitochondrial DNA (mtDNA) typically occurs as a single circular chromosome with 13 protein-coding genes and 22 tRNA genes. The various species of lice examined previously, however, have shown mitochondrial genome rearrangements with a range of chromosome sizes and numbers. Our research demonstrates that the mitochondrial genomes of two species of chewing lice found on pocket gophers, Geomydoecus aurei and Thomomydoecus minor, are fragmented with the 1,536 base-pair (bp) cytochrome-oxidase subunit I (cox1) gene occurring as the only protein-coding gene on a 1,916-1,964 bp minicircular chromosome in the two species, respectively. The cox1 gene of T. minor begins with an atypical start codon, while that of G. aurei does not. Components of the non-protein coding sequence of G. aurei and T. minor include a tRNA (isoleucine) gene, inverted repeat sequences consistent with origins of replication, and an additional non-coding region that is smaller than the non-coding sequence of other lice with such fragmented mitochondrial genomes. Sequences of cox1 minichromosome clones for each species reveal extensive length and sequence heteroplasmy in both coding and noncoding regions. The highly variable non-gene regions of G. aurei and T. minor have little sequence similarity with one another except for a 19-bp region of phylogenetically conserved sequence with unknown function.

New records of species of Saemundssonia (Insecta: Phthiraptera: Philopteridae) infesting breeding terns in the Arabian Peninsula, with notes on their phylogeny and ecology.

Six species of terns, which breed on the Arabian Peninsula, were examined for head chewing lice of the genus Saemundssonia in four different islands around the coasts of Saudi Arabia, both in the Red Sea and in the Arabian Gulf. Four louse species were collected: Saemundssonia laticaudata, Saemundssonia melanocephalus, Saemundssonia meridiana and Saemundssonia sternae, of which three are recorded for the first time from this region. Also, we record three new host-louse associations for the world-Saemundssonia laticaudata and Saemundssonia sternae from white-cheeked terns and Saemundssonia melanocephalus from Saunders's terns-including a host-switch event of Saemundssonia laticaudata on white-cheeked terns in the Karan Island population. Gene bank data for the COI gene from seven species of Saemundssonia that infest marine birds were used to propose evolutionary trees using two different statistical methods: maximum parsimony (MP) and neighbour joining (NJ). The result indicated that the tree which was produced by NJ is likely to be more accurate as it appeared more compatible with hosts' phylogeny. The trees indicate relationships between tern Saemundssonia and congeneric species from other marine birds, especially from gulls. An ANOVA was also conducted to test the mean parasite load for each tern species studied, and results indicate that there is a relation between louse loads and colonization behaviour of the hosts. Data from lice examined and illustrations of lice and their hosts are also included.

Characterization of 17 novel polymorphic microsatellite loci in the mammal chewing louse Geomydoecus ewingi (Insecta: Phthiraptera) for population genetic analyses.

We report 17 novel microsatellite loci in the parasitic chewing louse Geomydoecus ewingi, a common parasite of the pocket gopher, Geomys breviceps . Thirty-three G. ewingi individuals from 1 geographic locality and 3 pocket gopher hosts (populations) were genotyped at each locus. The number of alleles per locus ranged from 3 to 13. Observed heterozygosity ranged from 0.182 to 0.788. Four to 6 loci per louse population fell outside of Hardy-Weinberg expectations (HWE) and examination of population structure also revealed substantial homozygote excess as well as significant structure among louse populations. These findings are likely the consequence of biological characteristics of the lice (low dispersal abilities, population bottlenecks, etc.), which can result in inbreeding. Notably, when all louse individuals were analyzed together as 1 population, a Wahlund effect was detected, supporting that louse populations are restricted to 1 host individual. The microsatellite markers characterized in this study will be useful in future studies exploring the population dynamics in host-parasite systems, potentially yielding a better understanding of the processes underlying symbiotic associations.

Comparative ectoparasite loads of five seabird species in the Galapagos Islands.

In this paper we describe the ectoparasitic lice (Insecta: Phthiraptera) found on 5 species of seabirds (magnificent frigatebird Fregata magnificens; great frigatebird Fregata minor ; Nazca booby Sula granti ; blue-footed booby Sula nebouxii ; and red-footed booby Sula sula ) on the Galapagos Archipelago. We found 9 species of ectoparasitic lice: 5 species of Pectinopygus ischnocerans, 1 infesting each host; 2 species of Colpocephalum amblyceran lice, 1 on each frigatebird species; and 2 shared amblycerans, Eidmanniella albescens (Piaget, 1880) found on Nazca and blue-footed boobies and Fregatiella aurifasciata (Kellogg, 1899) found on the 2 frigatebirds. We tested the relative importance and interactions of host sex, body size, host, island, host family, and breeding status and found that inter-island differences were the main predictors of prevalence and infestation intensity. These differences could be related to host density or weather, but further evidence is needed.

Co-phylogeography and morphological evolution of sika deer lice (Damalinia sika) with their hosts (Cervus nippon).

Lice are obligate parasites of mammals and birds and have become an important model for studies of host-parasite co-evolution and co-phylogenetics. Population genetic and phylogeographic studies represent an important bridge between microevolution and co-phylogenetic patterns. We examine co-phylogeographic patterns in sika deer and their parasitic lice. Co-phylogeographic patterns in deer and lice were evaluated using homologous regions of mitochondrial COI sequences. The phylogeographic breaks recovered for deer populations matched those of previous studies. Comparisons of the phylogeographic tree topology for deer lice with that of their hosts revealed a significant level of congruence. However, comparisons of genetic distances between deer and lice suggested that one of the estimated co-divergence events is more likely a recent host switch. Taking into account genetic divergence, there is not strong evidence for complete phylogeographic co-divergence between deer and their parasitic lice. However, mitochondrial phylogenies only track genetic structure of female lineages, and the incongruence between deer and louse phylogeography may be explained by louse migration mediated by male deer. Morphological analysis of head shape variation based on an elliptic Fourier descriptor showed that overall morphological variation contained phylogenetic signal, suggesting that in general morphology of these lice evolves congruent to population history.

A new genus and species of Philopteridae (Phthiraptera: Ischnocera) from the trumpeters (Aves: Gruiformes: Psophiidae).

A new chewing louse genus and species belonging to the Philopteridae, namely, Palmaellus inexpectatus n. gen., n. sp., is described. The new genus is distinguished from the other ischnoceran genera hitherto described by its peculiar characters of the dorsal anterior head plate with 2 postero-lateral projections, pterothorax and abdomen with scarce chaetotaxy, male genitalia with simple mesomere and paramere lacking inner digitiform projection, and the genital region of female with postero-vulvar plates bearing setae. It is a parasite of the trumpeters, an avian family endemic to South America's Amazon Basin.

Cophylogeny on a fine scale: Geomydoecus chewing lice and their pocket gopher hosts, Pappogeomys bulleri.

Many species of pocket gophers and their ectoparasitic chewing lice have broadly congruent phylogenies, indicating a history of frequent codivergence. For a variety of reasons, phylogenies of codiverging hosts and parasites are expected to be less congruent for more recently diverged taxa. This study is the first of its scale in the pocket gopher and chewing louse system, with its focus entirely on comparisons among populations within a single species of host and 3 chewing louse species in the Geomydoecus bulleri species complex. We examined mitochondrial DNA from a total of 46 specimens of Geomydoecus lice collected from 11 populations of the pocket gopher host, Pappogeomys bulleri. We also examined nuclear DNA from a subset of these chewing lice. Louse phylogenies were compared with a published pocket gopher phylogeny. Contrary to expectations, we observed a statistically significant degree of parallel cladogenesis in these closely related hosts and their parasites. We also observed a higher rate of evolution in chewing louse lineages than in their corresponding pocket gopher hosts. In addition, we found that 1 louse species (Geomydoecus burti) may not be a valid species, that subspecies within G. bulleri are not reciprocally monophyletic, and that morphological and genetic evidence support recognition of a new species of louse, Geomydoecus pricei.

Flyway homogenisation or differentiation? Insights from the phylogeny of the sandpiper (Charadriiformes: Scolopacidae: Calidrinae) wing louse genus Lunaceps (Phthiraptera: Ischnocera).

The wing louse genus Lunaceps, is the most speciose chewing louse (Phthiraptera) genus inhabiting sandpipers (Charadriiformes: Calidrinae) and is known from almost all sandpiper species. The hosts follow specific flyways from the Arctic breeding grounds to wintering locations in the southern hemisphere, and often form large mixed-species flocks during migration and wintering. We estimated a phylogeny of Lunaceps based on three mitochondrial loci, supporting monophyly of the genus but revealing extensive paraphyly at the species level. We also evaluated the relative importance of flyway differentiation (same host species having different lice along different flyways) and flyway homogenisation (different host species having the same lice along the same flyway). We found that while the lice of smaller sandpipers and stints show some evidence of flyway homogenisation, those of larger sandpipers do not. No investigated host species migrating along more than one flyway showed any evidence of flyway differentiation. The host-parasite associations within Lunaceps are in no case monophyletic, rejecting strict cospeciation.

Evolution of cryptic coloration in ectoparasites.

Cryptic coloration is a classic example of evolution by natural selection. However, it has been studied almost exclusively in predator-prey systems, despite the fact that it may evolve in other groups, such as ectoparasites. The principle defense of hosts against ectoparasites is grooming behavior, which has a visual component. Host-imposed selection should lead to the evolution of background matching if it helps ectoparasites escape from grooming. Here we use sister taxa comparisons to show that avian feather lice (Phthiraptera: Ischnocera) have evolved coloration that matches the host's plumage, except in the case of head lice, which are protected from grooming. We also show covariation of parasite and host color within a single species of louse. Thus, cryptic coloration has evolved both within and between species of feather lice. Other examples of the evolution of crypsis presumably exist among the 70,000 known species of ectoparasites that collectively represent five animal phyla.