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.

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.

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.

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.

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.

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.

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.

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

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

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.

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

BACKGROUND: The mobility of birds across or between continents exposes them to numerous vectors that have the potential to transmit pathogens and spread them into new regions. A combination of rich species diversity of birds along with the small amount of molecular studies in Iran makes observing the blood parasite distribution in wild avian populations indispensable for further estimation and administration of blood parasites. METHODOLOGY/PRINCIPAL FINDINGS: In order to evaluate the infection rate and molecular context of avian blood parasites, bird samples were collected (passerine = 316 and non-passerine = 14) in eight provinces of northern Iran between June to September 2015 and 2016. All bird samples were examined for haematoprotozoan infections by morphological screening using light microscope and mtDNA cytb gene amplification. A total of 115 birds were positive for blood parasites by molecular approach (34.84% overall infection). The infection rate of Haemoproteus, Plasmodium, and Leucocytozoon were 33.03%, 1.21%, and 0.6%, respectively. Sequences analysis has detected 43 lineages in Iranian birds' hosts. Lineages were attributed to three genera Haemoproteus (n = 37), Plasmodium (n = 4), and Leucocytozoon (n = 2), of which 23 lineages fully matched previously recorded sequences in GenBank and MalAvi data reciprocities. Five lineages of ACDUM1, ACDUM2, PARUS1, PYERY01, and SISKIN1 were detected in multiple hosts' species from dissimilar families. In Bayesian tree, all sequences were clustered in three main monophyletic clades as Haemoproteus, Plasmodium, and Leucocytozoon genera. CONCLUSIONS/SIGNIFICANCE: As the first study outlining the molecular detection of hematozoa of passerines from Iran, the current study has recorded 20 new lineages for three genera of Haemoproteus, Plasmodium, and Leucocytozoon. Additional investigations into these taxa in the avifauna for the other parts of Iran may provide extra information on blood parasites, hosts relationships and distribution patterns.

Description of a new species of the genus Agamura Blanford, 1874 (Squamata: Gekkonidae) from southern Iran.

Herein, we describe a new species of spider gecko, Agamura kermanensis sp. nov., from south Kerman, Iran. The new species is distinguished from other species by having long forelimbs and hindlimbs and a high interorbital distance. Agamura kermanensis sp. nov. was discovered in the Faryab region of southern Kerman Province, which has a xeric climate and low vegetation density. The habitat of the species is similar to that of A. cruralis in southeast Iran. It can be assumed that the distribution of the species is more extensive than currently known. More investigation is required in Jazmourian and the surrounding areas. This is the third species of the genus Agamura discovered on the Iranian Plateau.

Occurrence of Haemoproteus spp. (Haemosporida: Haemoproteidae) in New Host Records of Passerine Birds from the East of Iran.

BACKGROUND: Avian haemosporidians are able to parasitize numerous bird species all over the world. The extensive range of blood parasites infection rate is between 50% and 100% or less percentage. Haemoparasites with major effects on physiology, ecology, health, population dynamics, sexual selection and production success of avian hosts may promote species extinction. METHODS: To evaluate haemosporidians infection rate in Iranian birds, 136 individuals were examined by microscopic observation of stained blood smears under light microscope. These samples belonged to 10 different families of Songbirds from the east of Iran from April to August 2014-2016. RESULTS: Fifty-one passerine birds were detected as harboring Haemoproteus spp. Furthermore, we recorded Haemoproteus spp. infection of Granativora bruniceps, Oenanthe pleschanka for the first time in the world and eight more species for Iran. CONCLUSION: Age and sampling localities do not influence the infection rate of Haemoproteus spp. from the eastern provinces of Iran. The relative high infection of avian haematozoa revealed this region might provide suitable sites for future studies on these parasites and the relationship with their hosts and vectors.

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.

Molecular Survey of Tularemia and Plague in Small Mammals From Iran.

Introduction: Plague and tularemia are zoonoses and their causative bacteria are circulating in certain regions of Iran. This study was conducted to investigate potential disease reservoirs amongst small wildlife species in different regions of Iran. Methods: Rodents, insectivores and hares from 17 different provinces of the country were collected in 2014 and 2015. Samples were taken from the spleens of the animals and Real-time PCR was applied to detect nucleic acid sequences that are specific to Francisella tularensis and Yersinia pestis, respectively. Results: Among 140 collected rodents, 25 distinct species were identified out of which five were the most common: Microtus paradoxus (21% out of 140 rodents), Apodemus witherbyi (12%), Microtus irani (11%), Mus musculus (11%) and Microtus socialis (10%). Seventeen insectivores were collected and identified as Crocidura suaveolens (82%) and C. leucodon (18%). Fifty-one hares were collected and identified as Lepus europaeus (57%), Lepus tolai (14%) and Lepus sp. (29%). Three out of 140 explored rodents (1.91%) were positive for F. tularensis, an A. witherbyi, a Mus musculus domesticus, and a Chionomys nivalis collected from Golestan, Khuzestan and Razavi Khorasan provinces, respectively. Two hares (3.92%) were F. tularensis-positive, a L. europaeus from Khuzestan and a Lepus sp. from the Sistan and Baluchistan province. None of the tested animals were positive for Y. pestis. Conclusion: This is the first report of direct detection of F. tularensis in mammals of Iran and the first-time observation of the agent in a snow vole, C. nivalis worldwide. The results indicate that tularemia is more widespread in Iran than previously reported including the Northeast and Southwestern parts of the country. Future studies should address genetic characterization of F. tularensis positive DNA samples from Iran to achieve molecular subtyping and rule out assay cross-reactivity with near neighbor Francisella species.

A new Pardosa species from northern Iran (Araneae, Lycosidae).

A study of Pardosa pontica (Thorell, 1875) in Iran indicates two distinct morphospecies. A detailed comparison of these morphospecies and types of P. pontica reveals that populations from Mazandaran, Golestan and North Khorasan Provinces of Iran and Balkan Welaýaty of Turkmenistan belong to a newly described species, P. mirzakhaniae sp. n., and populations in western and easternmost Iran belong to P. pontica. Although the two morphospecies differ in colour, size and spination, no molecular differences in 16S rRNA and COI were detected.

First report of bioaccumulation and bioconcentration of aliphatic hydrocarbons (AHs) and persistent organic pollutants (PAHs, PCBs and PCNs) and their effects on alcyonacea and scleractinian corals and their endosymbiotic algae from the Persian Gulf, Iran: Inter and intra-species differences.

The coral reefs of the Persian Gulf are the most diverse systems of life in the marine environment of the Middle East. Unfortunately, they are highly threatened by local and global stressors, particularly oil pollutants. This is the first quantitative and qualitative study aimed at assessing the concentration and sources of n-alkanes and POPs (PAHs, PCBs and PCNs) in coral tissues, symbiotic algae (zooxanthellae), reef sediments and seawaters in coral reefs of Lark and Kharg in the Persian Gulf, Iran. This work was conducted on eight species of six genera and three families of hard corals and one family of soft coral. A significant variation in the concentration of ∑30n-alkanes and POPs (∑40PAHs, ∑22PCBs and 20PCNs) was found in the decreasing order: zooxanthellae > coral tissue > skeleton > reef sediment > seawater. The bioaccumulation of these compounds was 2-times higher in ahermatypic than in hermatypic corals, among which significant variations were observed in both sites. In Kharg, Porites lutea had the highest mean concentration of ∑30n-alkanes and ∑40PAHs in soft tissue, whereas the lowest values were in Platygyra daedalea. A contrasting trend was documented for ∑22PCBs and 20PCNs, with the highest level reported in soft tissue of P. daedalea and the lowest in P. lutea at Kharg. Compositional pattern of AHs and PAHs demonstrated the predominance of LMW-PAHs and n-alkanes. In skeleton and reef sediments, tetra, penta and tri-CBs were the most abundant PCBs congeners followed by di-CB > hexa-CB > hepta-CB > octa-CB,whiletri-CB > di-CB > tetra-CB > penta-CB > hexa-CB > hepta-CB > octa-CB was observed for soft tissue, zooxanthellae and seawater. The results of RAD test indicated significantly negative correlation between total concentration of these compounds with zooxanthellae density, the chlorophyll-a and C2 in corals at both reefs. This is the first report on levels, health assessment and source apportionments of POPs in zooxanthellae and a first step in the implementation of specific coral reef management measures.

New Host Records for Haemoproteus Spp. (Apicomplexa: Haemosporidiasina) in Passeriformes from North-West of Iran.

BACKGROUND: The intracellular protozoan parasites of the genus Haemoproteus occur in different avian hosts all over the world. Various genus of blood sucking insects' families such as Hippoboscidae and Ceratopogonidae could transmit Haemoproteus in avian hosts. There are very limited number of studies on wild infected birds with blood parasites in Iran, so the aim of this study was to determine the frequency of Haemoproteus spp. infection in passerine birds from northwest of Iran. METHODS: Passerines were collected from four different localities in Zanjan Province, northwest Iran during June to August 2014. RESULTS: Of 86 passerines, we found Haemoproteus infection in 19 (22.09%) individuals. In general, 15 bird species were observed for haemosporidians, of which 53% were infected. CONCLUSION: Three species of passerines: Petronia petronia, Sitta tephronota and Acrocephalus melanopogon are new host records for Haemoproteus infection in the world. Results acquired by this study support widespread distribution of Haemoproteus in passerines and illustrated the prevalence of Haemoproteus species in wild birds of northwest of Iran. Conclusively, our study specified that more investigations are needed to reach exact prevalence rate in different families of birds in Iran.

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.