Phylogeny and evolutionary history of the Sombre Tit, Poecile lugubris in the western Palearctic (Aves, Paridae).

Few studies have delimited evolutionary entities within the Sombre Tit, Poecile lugubris. Here, we explored its morphological and genetic variability using 24 morphometric variables, two mitochondrial (COX1 and ND2), two nuclear (ODC and MB), and 10 microsatellite loci. Genetic and morphometric characters supported the species status of the Caspian Tit, Poecile hyrcanus with a separation from P. lugubris more than 4.5 MYA. The phylogenetic analyses uncovered three distinct clades within P. lugubris. The subspecies P. l. lugubris, with strong genetic differences from the other subspecies (4.5%), diverged at ∼1.1 MYA. Samples from Iran formed the remaining two clades. Individuals from western and northwestern Iran were placed in a single clade (anatoliae), while those from southern and southwestern Iran were in another independent clade (dubius-kirmanensis). Morphometric analyses also confirmed this pattern. The microsatellite results discriminated Iranian subspecies as discrete clusters with signs of nuclear admixture between dubius and anatoliae in the Zagros Mountains. Signs of population expansion for anatoliae and dubius-kirmanensis coincided with the late LGM. Our results shed new light on the phylogenetic relationships, evolutionary history, and past demographic processes of P. lugubris.

Multiple species delimitation approaches applied to the avian lark genus Alaudala.

Species delimitation has advanced from a purely phenotypic exercise to a branch of science that integrates multiple sources of data to identify independently evolving lineages that can be treated as species. We here test species limits in the avian Lesser Short-toed Lark Alaudala rufesens-Sand Lark A. raytal complex, which has an intricate taxonomic history, ranging from a single to three recognised species, with different inclusiveness in different treatments. Our integrative taxonomic approach is based on a combination of DNA sequences, plumage, biometrics, songs, song-flights, geographical distributions, habitat, and bioclimatic data, and using various methods including a species delimitation program (STACEY) based on the multispecies coalescent model. We propose that four species should be recognised: Lesser Short-toed Lark A. rufescens (sensu stricto), Heine's Short-toed Lark A. heinei, Asian Short-toed Lark A. cheleensis and Sand Lark A. raytal. There is also some evidence suggesting lineage separation within A. cheleensis and A. raytal, but additional data are required to evaluate this. The species delimitation based on STACEY agrees well with the non-genetic data. Although computer-based species delimitation programs can be useful in identifying independently evolving lineages, we stress that whenever possible, species hypotheses proposed by these programs should be tested by independent, non-genetic data. Our results highlight the difficulty and subjectivity of delimiting lineages and species, especially at early stages in the speciation process.

Genome-wide evidence supports mitochondrial relationships and pervasive parallel phenotypic evolution in open-habitat chats.

In wheatears and related species ('open-habitat chats'), molecular phylogenetics has led to a comprehensively revised understanding of species relationships and species diversity. Phylogenetic analyses have suggested that, in many cases, phenotypic similarities do not reflect species' relationships, revealing traditionally defined genera as non-monophyletic. This led to the suggestion of pervasive parallel evolution of open-habitat chats' plumage coloration and ecological phenotypes. However, to date, the molecular evidence for the phylogenetic relationships among open-habitat chats is mainly limited to mitochondrial DNA. Here, we assessed whether the mitochondrial relationships are supported by genome-wide data. To this end, we reconstructed the species tree among 14 open-habitat chat taxa using multi-species coalescent analyses based on ~1'300 SNPs. Our results confirm previous ones based chiefly on mitochondrial DNA; notably the paraphyly of the Oenanthe lugens complex and the clustering of individual species formerly placed in the genera Cercomela and Myrmecocichla within Oenanthe. Since several variable morphological and ecological characteristics occur in multiple places across the open-habitat chat phylogeny, our study consolidates the evidence for pervasive parallel evolution in the plumage coloration and ecology of open-habitat chats.

Parallel plumage colour evolution and introgressive hybridization in wheatears.

Genetic and phenotypic mosaics, in which various phenotypes and different genomic regions show discordant patterns of species or population divergence, offer unique opportunities to study the role of ancestral and introgressed genetic variation in phenotypic evolution. Here, we investigated the evolution of discordant phenotypic and genetic divergence in a monophyletic clade of four songbird taxa-pied wheatear (O. pleschanka), Cyprus wheatear (Oenanthe cypriaca), and western and eastern subspecies of black-eared wheatear (O. h. hispanica and O. h. melanoleuca). Phenotypically, black back and neck sides distinguish pied and Cyprus wheatears from the white-backed/necked black-eared wheatears. Meanwhile, mitochondrial variation only distinguishes western black-eared wheatear. In the absence of nuclear genetic data, and given frequent hybridization among eastern black-eared and pied wheatear, it remains unclear whether introgression is responsible for discordance between mitochondrial divergence patterns and phenotypic similarities, or whether plumage coloration evolved in parallel. Multispecies coalescent analyses of about 20,000 SNPs obtained from RAD data mapped to a draft genome assembly resolve the species tree, provide evidence for the parallel evolution of colour phenotypes and establish western and eastern black-eared wheatears as independent taxa that should be recognized as full species. The presence of the entire admixture spectrum in the Iranian hybrid zone and the detection of footprints of introgression from pied into eastern black-eared wheatear beyond the hybrid zone despite strong geographic structure of ancestry proportions furthermore suggest a potential role for introgression in parallel plumage colour evolution. Our results support the importance of standing heterospecific and/or ancestral variation in phenotypic evolution.

Signatures of human-commensalism in the house sparrow genome.

House sparrows (Passer domesticus) are a hugely successful anthrodependent species; occurring on nearly every continent. Yet, despite their ubiquity and familiarity to humans, surprisingly little is known about their origins. We sought to investigate the evolutionary history of the house sparrow and identify the processes involved in its transition to a human-commensal niche. We used a whole genome resequencing dataset of 120 individuals from three Eurasian species, including three populations of Bactrianus sparrows, a non-commensal, divergent house sparrow lineage occurring in the Near East. Coalescent modelling supports a split between house and Bactrianus sparrow 11 Kya and an expansion in the house sparrow at 6 Kya, consistent with the spread of agriculture following the Neolithic revolution. Commensal house sparrows therefore likely moved into Europe with the spread of agriculture following this period. Using the Bactrianus sparrow as a proxy for a pre-commensal, ancestral house population, we performed a comparative genome scan to identify genes potentially involved with adaptation to an anthropogenic niche. We identified potential signatures of recent, positive selection in the genome of the commensal house sparrow that are absent in Bactrianus populations. The strongest selected region encompasses two major candidate genes; COL11A-which regulates craniofacial and skull development and AMY2A, part of the amylase gene family which has previously been linked to adaptation to high-starch diets in humans and dogs. Our work examines human-commensalism in an evolutionary framework, identifies genomic regions likely involved in rapid adaptation to this new niche and ties the evolution of this species to the development of modern human civilization.

Natural epigenetic variation within and among six subspecies of the house sparrow, Passer domesticus.

Epigenetic modifications can respond rapidly to environmental changes and can shape phenotypic variation in accordance with environmental stimuli. One of the most studied epigenetic marks is DNA methylation. In the present study, we used the methylation-sensitive amplified polymorphism (MSAP) technique to investigate the natural variation in DNA methylation within and among subspecies of the house sparrow, Passer domesticus We focused on five subspecies from the Middle East because they show great variation in many ecological traits and because this region is the probable origin for the house sparrow's commensal relationship with humans. We analysed house sparrows from Spain as an outgroup. The level of variation in DNA methylation was similar among the five house sparrow subspecies from the Middle East despite high phenotypic and environmental variation, but the non-commensal subspecies was differentiated from the other four (commensal) Middle Eastern subspecies. Further, the European subspecies was differentiated from all other subspecies in DNA methylation. Our results indicate that variation in DNA methylation does not strictly follow subspecies designations. We detected a correlation between methylation level and some morphological traits, such as standardized bill length, and we suggest that part of the high morphological variation in the native populations of the house sparrow is influenced by differentially methylated regions in specific loci throughout the genome. We also detected 10 differentially methylated loci among subspecies and three loci that differentiated between commensal or non-commensal status. Therefore, the MSAP technique detected larger scale differences among the European and non-commensal subspecies, but did not detect finer scale differences among the other Middle Eastern subspecies.

Avian gametologs as molecular tags for sex identification in birds of prey of Iran.

Global environmental change and rapid destruction of natural habitats necessitate the conservation of endangered and threatened birds of prey. Recently, molecular sex identification methods based on amplification of introns of chromodomain-helicase DNA binding protein1 (CHD1) have provided valuable tools for ecological study and conservation breeding programs of birds. These methods employ a primer pair flanking an intron which varies considerably in length between the avian gametologs CHD1Z and CHD1W. Herein, we test the applicability of CHD1Z and CHD1W as universal tags for molecular sex identification in birds of prey of Iran. We showed successful sex identification in 22 species of birds of prey using feathers as the source of DNA. The results suggest that the regions of CHD1W and CHD1Z amplified in this study are conserved among most of Falconiformes, enabling accurate sex identification in birds of prey.

Mercury contamination in Khramulia (Capoeta capoeta) from the Cheshme Kile and Zarrin Gol Rivers in Iran and human health risk assessment.

Total mercury concentrations were determined in muscle tissue of Khramulia (Capoeta capoeta) captured in the Cheshme Kile and Zarrin Gol Rivers, Iran. In Cheshme Kile River, 49 fish samples were collected. The mean total mercury concentration in the muscles of C. capoeta from this area was 249 ng g(-1) dw. In Zarrin Gol River, where 62 fish samples were collected, the total mercury in muscles averaged 164 ng g(-1) dw. A significant difference was found between means of mercury in the rivers (p < 0.001). All samples from the two rivers had mean mercury concentrations below the maximum allowable limits for mercury set by the Food and Agriculture Organization, World Health Organization, Standardization Administration of China and Environmental Protection Agency. The results of this study indicate that the values of hazard target quotient and estimated weekly intake are low and represent a negligible risk for human health.

Beak and skull shapes of human commensal and non-commensal house sparrows Passer domesticus.

BACKGROUND: The granivorous house sparrow Passer domesticus is thought to have developed its commensal relationship with humans with the rise of agriculture in the Middle East some 10,000 years ago, and to have expanded with the spread of agriculture in Eurasia during the last few thousand years. One subspecies, P. d. bactrianus, residing in Central Asia, has apparently maintained the ancestral ecology, however. This subspecies is not associated with human settlements; it is migratory and lives in natural grass- and wetland habitats feeding on wild grass seeds. It is well documented that the agricultural revolution was associated with an increase in grain size and changes in seed structure in cultivated cereals, the preferred food source of commensal house sparrow. Accordingly, we hypothesize that correlated changes may have occurred in beak and skull morphology as adaptive responses to the change in diet. Here, we test this hypothesis by comparing the skull shapes of 101 house sparrows from Iran, belonging to five different subspecies, including the non-commensal P. d. bactrianus, using geometric morphometrics. RESULTS: The various commensal house sparrow subspecies share subtle but consistent skeletal features that differ significantly from those of the non-commensal P. d. bactrianus. Although there is a marked overall size allometry in the data set, the shape difference between the ecologically differentiated sparrows cannot be explained by differences in size alone. Relative to the size allometry commensal house sparrows exhibit a skull shape consistent with accelerated development (heterochrony), resulting in a more robust facial cranium and a larger, more pointed beak. CONCLUSION: The difference in skull shape and robustness of the beak between commensal and non-commensal house sparrows is consistent with adaptations to process the larger and rachis encapsulated seeds of domesticated cereals among human associated populations.

DNA barcoding as a tool for elucidating species delineation in wide-ranging species as illustrated by owls (Tytonidae and Strigidae).

The mitochondrial cytochrome c-oxidase subunit I (cox1) can serve as a fast and accurate marker for the identification of animal species, and for the discovery of new species across the tree of life. Distinguishing species using this universal molecular marker, a technique known as DNA barcoding, relies on the identifying the gap between intra- and interspecific divergence. One of the difficulties could be wide-ranging, cosmopolitan species that show large amounts of morphological variation. The barn owl Tyto alba is a case in point. It occurs worldwide and varies morphologically, leading to the recognition of many subspecies or, more recently, species. We analysed data from the cox1 gene for 31 individuals of seven subspecies, and compared this with 214 sequences from 29 other owl species. Phylogenetic analysis of the T. alba samples gives very strong support for an Old World alba-clade (three subspecies) and a New World furcata-clade (four subspecies) that are genetically equidistant. The amount of intraspecific variation within each of these clades ranges from 0.66-0.99%, but variation among these clades ranges from 5.33-6.20%. Combined these data suggest that barn owl of the Old World is indeed best considered a separate species different from that of the New World. For combined dataset, sample size of owl species (n between 1 and 21 sequences) increased with geographic range size but we did not find significant relationships between interspecific divergence and sample size or between interspecific divergence and geographic range. For 21/24 species of owls with sample sizes of n ≥4 the maximum interspecific divergences was ≤ 3.00%. However, similar to those found in barn owls, the largest amount of divergence (3.23-4.09%) was present in two other wide-ranging species (Strix nebulosa and Aegolius funereus) raising the possibility of multiple species in other wide-ranging owls as well.

Odontobuthus tirgari sp. nov. (Scorpiones, Buthidae) from the eastern region of the Iranian Plateau .

A new species of scorpions in the genus Odontobuthus (Scorpiones, Buthidae) is described from Khorasan Province, Iran. Currently, Odontobuthus includes two species in Iran, Odontobuthus doriae Thorell, 1876, which is restricted to high elevations of the central Iranian Plateau and Odontobuthus bidentatus Lourenço & Pezier, 2002 from the Zagros Mountains. The results of morphological comparisons, univariate and multivariate statistical analyses and phylogenetic analysis of COI sequence data clearly confirm a deep split between populations from the eastern Iranian Plateau and O. bidentatus Lourenço & Pezier, 2002 and O. doriae Thorell, 1876. Therefore, according to comparative morphological and molecular analyses, a new species, Odontobuthus tigari sp. nov. (♀♂) was described from eastern Iran. This addition represents the third species of this genus from Iran.

Convergent evolution of morphological and ecological traits in the open-habitat chat complex (Aves, Muscicapidae: Saxicolinae).

Open-habitat chats (genera Myrmecocichla, Cercomela, Oenanthe and relative) are a morphologically and ecologically cohesive group of genera with unclear phylogenetic relationships. They are distributed mostly in open, arid and/or rocky habitats of Africa and Eurasia. Here, we present the most comprehensive molecular phylogenetic analysis of this group to date, with a complete taxon sampling at the species level. The analysis, based on a multilocus dataset including three mitochondrial and three nuclear loci, allows us to elucidate the phylogenetic relationships and test the traditional generic limits. All genera are non-monophyletic, suggesting extensive convergence on similar plumage patterns in unrelated species. While the colour pattern appear to be a poor predictor of the phylogenetic relationships, some of the ecological and behavioural traits agree relatively well with the major clades. Following our results, we also propose a revised generic classification for the whole group.

Prevalence of co-infection and genetic diversity of avian haemosporidian parasites in two rehabilitation facilities in Iran: implications for the conservation of captive raptors.

BACKGROUND: Various haemosporidian parasites infect raptors, especially captive hosts who may be more exposed. Diagnosis of threatening factors such as infectious diseases indirectly has a significant role in protecting endangered or threatened species that may boost the mortality or extinction resulting from declined reproduction. Few investigations have been performed in captive hosts to detect the prevalence of haemosporidian parasites and define genetic diversity in west Asia. For the first time, the current study was designed to determine the prevalence and genetic diversity of haemosporidian parasites in captive raptors by molecular methods in two rehabilitation facilities in North and North-east Iran and to define phylogenetic relationships of detected lineages circulating in raptors. RESULTS: Molecular characterization of the haemosporidian parasite was accomplished by PCR-based method and DNA sequencing in 62 captive raptors. The overall prevalence was ~ 36% with higher infection of Haemoproteus spp. than Leucocytozoon spp. Plasmodium infection was not detected in any host. Results showed that 22 individuals (of 10 species) were infected with unique lineages. Genus Haemoproteus was detected in 26.66% of examined individuals (of eight species) and Leucocytozoon was found in 10% of individuals (of four species). The molecular analysis could detect ten lineages (nine Haemoproteus spp. and one Leucocytozoon spp.) which were categorizes as new and six lineages which have been previously detected in the other investigations. CONCLUSIONS: The Bayesian phylogenetic analysis derived from obtained data in the present study and published lineages in previous investigations indicated the probable host specificity of Haemoproteus and Leucocytozoon parasites in several sub-clades at hosts' order and genus level. As monitoring the parasite loads of captive birds when admitted reduce the risk of infecting hosts in captivity at those locations, we designed this study to determine infection prevalence and genetic diversity of blood parasites in raptors examined in Iran. These results allow mapping of haemosporidian distribution and shed light on the depth of their diversity in Iran to protect species by identification of risk in rehabilitation facilities.

Prevalence and Genetic Diversity of Avian Haemosporidian Parasites in Southern Iran.

Avian haemosporidians are widespread and diverse and are classified in the genera Plasmodium, Haemoproteus, Leucocytozoon, and Fallisia. These species are known to cause haemosporidiosis and decreased fitness of their hosts. Despite the high diversity of habitats and animal species in Iran, only few studies have addressed avian haemosporidians in this geographic area. This study was performed in the south and southeast of Iran during the bird breeding seasons in 2017 and 2018, with the aim to partly fill in this gap. Blood samples of 237 passerine birds belonging to 41 species and 20 families were collected. Parasite infections were identified using a nested PCR protocol targeting a 479-base-pair fragment of the mitochondrial cytochrome b (cytb) gene of Haemoproteus, Plasmodium and Leucocytozoon species. The overall prevalence of haemosporidian parasites was 51.1%, and 55 different lineages were identified, of which 15 cytb lineages were new globally. The lineages of Haemoproteus predominated (63.6% of all detected lineages), followed by Leucocytozoon and Plasmodium. Nineteen new host records of haemosporidian cytb lineages were identified, and the majority of them were found in resident bird species, indicating local transmission. Thirteen co-infections (9.8% of infected individuals) of Haemoproteus and Leucocytozoon parasites in seven host species were observed. This study shows the presence of active local transmission of parasites to resident bird species in the southeast of Iran and contributes to the knowledge on haemosporidian parasite biodiversity in this poorly studied region of the world.

New Data on Comparative Cytogenetics of the Mouse-Like Hamsters (Calomyscus Thomas, 1905) from Iran and Turkmenistan.

The taxonomy of the genus Calomyscus remains controversial. According to the latest systematics the genus includes eight species with great karyotypic variation. Here, we studied karyotypes of 14 Calomyscus individuals from different regions of Iran and Turkmenistan using a new set of chromosome painting probes from a Calomyscus sp. male (2n = 46, XY; Shahr-e-Kord-Soreshjan-Cheshme Maiak Province). We showed the retention of large syntenic blocks in karyotypes of individuals with identical chromosome numbers. The only rearrangement (fusion 2/21) differentiated Calomyscus elburzensis, Calomyscus mystax mystax, and Calomyscus sp. from Isfahan Province with 2n = 44 from karyotypes of C. bailwardi, Calomyscus sp. from Shahr-e-Kord, Chahar Mahal and Bakhtiari-Aloni, and Khuzestan-Izeh Provinces with 2n = 46. The individuals from Shahdad tunnel, Kerman Province with 2n = 51-52 demonstrated non-centric fissions of chromosomes 4, 5, and 6 of the 46-chromosomal form with the formation of separate small acrocentrics. A heteromorphic pair of chromosomes in a specimen with 2n = 51 resulted from a fusion of two autosomes. C-banding and chromomycin A3-DAPI staining after G-banding showed extensive heterochromatin variation between individuals.

The Lanius excubitor (Aves, Passeriformes) conundrum--Taxonomic dilemma when molecular and non-molecular data tell different stories.

The phylogeny of 18 taxa in the Lanius excubitor complex, and the related species L. sphenocercus, L. ludovicianus and L. somalicus, was estimated based on the mitochondrial cytochrome b gene and the non-coding D-loop (in total approximately 1.3 kb). According to the mitochondrial gene tree, Lanius excubitor s.l. is non-monophyletic, with some of its subspecies being more closely related to L. sphenocercus, L. ludovicianus, and L. somalicus. Also the division of the L. excubitor complex into a northern (L. excubitor) and a southern (L. meridionalis) species, as has been proposed based on morphological and ecological similarity and geographical distributions, is not compatible with the mitochondrial tree. Overall, genetic divergences among the ingroup taxa are small, indicating a recent radiation. A tree based on the nuclear ornithine decarboxylase (ODC) introns 6-7 is unresolved with respect to the ingroup, but provides strong support for a clade containing the Lanius excubitor complex, L. sphenocercus, L. ludovicianus and L. somalicus. We discuss the incongruence between the current taxonomy and the mitochondrial gene tree, and conclude that based on the latter the Lanius excubitor complex may be treated as at least six species, L. borealis, L. elegans, L. excubitor, L. lahtora, L. meridionalis, and L. uncinatus, but that other taxonomic treatments are also possible. However, uncertainty regarding to which extent the mitochondrial gene tree reflects the species phylogeny prevents us from recommending taxonomic change without further investigation. This study highlights the possible danger of relying on a single molecular marker, such as mitochondrial DNA, in taxonomic revisions and phylogenetic inference.

Phylogeny of the mourning wheatear Oenanthe lugens complex.

The phylogenetic relationship of many species and subspecies within the genus Oenanthe (wheatears) is still debated. Only recently molecular approaches have been used to clarify their basal taxonomy. One of the main unsolved groups is summarized under the name mourning wheatear O. lugens, which comprises depending on the underlying species concept 3-8 different taxa. These include the wheatears of the subspecies group lugens (halophila, lugens, persica), the subspecies group lugubris (lugubris, schalowi, vauriei), and the subspecies group lugentoides (lugentoides, boscaweni). In order to shed light on this unsolved issue we studied the taxonomy of the mourning wheatear complex by means of molecular markers and comparative morphometry. We found reasonable evidence to follow a narrow species concept treating all the three subspecies groups of the mourning wheatear in future as three independent taxonomic entities (super-species O. lugens, O. lugentoides, O. lugubris). Further within the subspecies group O. lugens we suggest treating halophila and lugens as members of the polytypic species O. lugens, while the Persian mourning wheatear O. persica merits the status of an independent monotypic species, endemic to the Iranian Plateau. Genetic and morphometric characters support a long separation of this form (a half to one Million years). For the black form "basalti" from Syria and Jordan our molecular data indicates a close relationship to lugens and this form is therefore probably best treated as a colour morph of lugens, adapted to the local habitat conditions. However, future behavioural studies have to show if there exist prezygotic barriers between both forms.

TMTCPT: The Tree Method based on the Taxonomic Categorization and the Phylogenetic Tree for fine-grained categorization.

Fine-grained categorization is one of the most challenging problems in machine vision. Recently, the presented methods have been based on convolutional neural networks, increasing the accuracy of classification very significantly. Inspired by these methods, we offer a new framework for fine-grained categorization. Our tree method, named "TMTCPT", is based on the taxonomic categorization, phylogenetic tree, and convolutional neural network classifiers. The word "taxonomic" has been derived from "taxonomical categorization" that categorizes objects and visual features and performs a prominent role in this category. It presents a hierarchical categorization that leads to multiple classification levels; the first level includes the general visual features having the lowest similarity level, whereas the other levels include visual features strikingly similar, as they follow top-bottom hierarchy. The phylogenetic tree presents the phylogenetic information of organisms. The convolutional neural network classifiers can classify the categories precisely. In this study, the researchers created a tree to increase classification accuracy and evaluated the effectiveness of the method by examining it on the challenging CUB-200-2011 dataset. The study results demonstrated that the proposed method was efficient and robust. The average classification accuracy of the proposed method was 88.34%, being higher than those of all the previous methods.

New insights to the taxonomy of Rhagodes eylandti (Walter, 1889): A remarkable sexually dimorphic species (Solifugae: Rhagodidae).

The taxonomy of the widespread camel spider, Rhagodes eylandti (Walter, 1889), is herein updated and revised by proposing three nominal taxa as its junior synonyms. These are based on data from males of two taxa, Rhagodes melanopygus nigricans Birula, 1905 and R. plumbescens (Walter, 1889), and a female of R. melanochaetus Heymons, 1902. Consequently, both sexes of R. eylandti are re-described and the validity of their morphological diagnostic characters is evaluated. Detailed morphological and morphometrical characters, as well as data on sexual dimorphism and intraspecific variations, are provided. The illustrations of type material are given for the first time. Moreover, a distribution map and ecological notes are presented.

Correction: Molecular detection and genetic diversity of avian haemosporidian parasites in Iran.

[This corrects the article DOI: 10.1371/journal.pone.0206638.].