mtDNA Gene Sequences Research Articles

Hidden in the frog: morphological and molecular characterization of Halipegus ovocaudatus (Platyhelminthes: Trematoda), type species of the genus Halipegus Looss, 1899.

The type species of the genus Halipegus Looss, 1899, Halipegus ovocaudatus (Vulpian, 1859) is a generalist parasite of different species of amphibians, mainly anurans. To date, the phylogenetic position of this species has not been established. Specimens of H. ovocaudatus, were collected from three species of water frogs, Pelophylax ridibundus (Pallas), P. esculentus (Linnaeus), and P. lessonae (Camerano) inhabiting the Middle Volga region (European Russia). In our study, we provide detailed morphological redescription of these digeneans and combine it with molecular phylogenetic analyses based on partial sequences of the 28S rDNA and coI mtDNA genes. The phylogenetic position of H. ovocaudatus among the derogenids has been established as a sister clade to other representatives of the Halipeginae. We compared pairwise distances based on partial sequences of coI mtDNA genes of H. ovocaudatus and other representatives of Derogenidae. It was shown, that adults of H. ovocaudatus from different hosts are representatives of the same species. Specimens parasitizing three frog hosts from several localities differ slightly in body shape and size, which probably reflects host-induced intraspecific variability.

Genetic Diversity of Bluefish Population in Türkiye Coastal

Bluefish (Pomatomus saltatrix) is a commercially important, highly migratory predatory fish species distributed along the coasts of Türkiye. Conservation and sustainability of bluefish populations requires knowledge of the genetic structure of the population, but There are no genetic studies on bluefish in Türkiye. In this study, the population structure of Bluefish in the coastal regions of Türkiye was investigated by sequence analysis of mitochondrial DNA gene regions (Cytochrome b and Displacement-loop) and nuclear gene region (Rhodopsin). We tried to identify bluefish samples collected from 14 regions in the coastal region of Türkiye by analyzing two mtDNA gene sequences (Cyt b and D-loop) and nuclear gene region (Rhodopsin). While a total of 35 haplotypes were found in the Cytb gene region, a total of 132 haplotypes were found in the D-loop (control region) gene region. A single haplotype was detected in all stations for Rhodopsin. Also, the maximum likelihood trees based on Cyt b and D-loop sequences suggested relationship of Bluefish populations from Türkiye and the world. Results showed a low level of genetic differentiation between Bluefish populations in the study area, but Aegean and Mediterranean populations are more diverse than other populations due to migratory trait of this species.

The first Genetic data on <i>Lernaea Cyprinacea</i> Linnaeus, 1758 from the European North of Russia (The Pinega River)

Morphological and molecular genetic data about Lernaea cyprinacea Linnaeus, 1758, collected from pike (Esox lucius Linnaeus, 1758) in the European North of Russia (Pinega river) in 2022, were presented. Molecular genetic studies were carried out based on the analysis of COI mtDNA, 18S rDNA and 28S rDNA gene sequences. A significant genetic distance for the COI gene from individuals of the same species collected in other territories (China, Australia, South Africa, Canada, Austria, Argentina, Pakistan, Iran, India, Japan, Romania, Egypt and Western Siberia) was revealed, which indicates the existence of a cryptic species L. cyprinacea in the European North of Russia. Analysis of 28S rDNA sequences showed the presence of two substitutions fixed in the population, which may indicate a fairly long-standing divergence of sister species. The 18S rDNA gene fragment is less suitable for identifying L. cyprinacea due to low variability compared to the other two gene regions.

Is the Javan tiger Panthera tigris sondaica extant? DNA analysis of a recent hair sample

Abstract The Javan tiger Panthera tigris sondaica and the Bali tiger P. tigris balica were categorized as Extinct on the IUCN Red List in both 2003 and 2008, leaving only the Sumatran subspecies P. tigris sumatrae extant in Indonesia. There have, however, been occasional, more recent reports of the Javan tiger but without conclusive evidence. Here, a potential observation in 2019 of a Javan tiger in a community plantation near the village of Cipendeuy in the forest of South Sukabumi, West Java, and a single hair found on a fence nearby, are assessed. The cytochrome b mitochondrial DNA (mtDNA) gene sequence of the putative Javan tiger hair were compared with that of a Javan tiger specimen in Museum Zoologicum Bogoriense, collected in 1930; hair samples of several tiger subspecies and the Javan leopard Panthera pardus melas were used as controls. The results showed that the genetic distances (d) of the putative Javan tiger hair with the Sumatran, Bengal P. tigris tigris and Amur P. tigris altaica tigers and the Javan leopard are 0.074 ± SE 0.009, 0.071 ± SE 0.009, 0.072 ± SE 0.009 and 0.088 ± SE 0.010, respectively, whereas the genetic distance of the putative Javan tiger hair with the Javan tiger museum specimen is 0.040 ± SE 0.006. In addition, phylogenetic trees showed that the putative Javan tiger hair sample belongs to the same group as the museum specimen of the Javan tiger, but is differentiated from other tiger subspecies and the Javan leopard. Whether the Javan tiger still occurs in the wild needs to be confirmed with further genetic and field studies.

Molecular characterization of the camel nasal botfly, Cephalopina titillator (Diptera: Oestridae)

BackgroundLarval stages of the camel nasal botfly, Cephalopina titillator (Diptera: Oestridae), rank among the top obligate endoparasites causing nasopharyngeal myiasis. Such infestations have adverse effects on camel health, reducing their economic value. The current study’s objective is to assess the applicability of PCR using partial regions of the COI mtDNA and 28S rRNA gene sequences to identify, classify, and reveal the phylogenesis of C. titillator.ResultsLarvae of the 2nd and 3rd instars of C. titillator were collected from the El-Bassatin abattoir in Egypt, extracted from the nasopharyngeal passage of the Arabian camel (Camelus dromedarius) during postmortem inspection. Genomic DNA was successfully isolated from 10 samples of 3rd instar larvae of C. titillator and amplified using primer pairs targeting partial mitochondrial COI (916 bp) and ribosomal 28S rRNA (830 bp) gene fragments. Nucleotide sequences from five samples have been sequenced and submitted to GenBank under accession numbers OP482168 to OP482172 for the COI gene and OP482160 to OP482164 for the 28S rRNA gene fragments. The COI gene exhibited 97.3% nucleotide identity across all specimens, while the 28S rRNA gene displayed 99.74% identical nucleotides. Maximum likelihood trees were constructed based on the generalized time-reversible (GTR) model. The resulting COI phylogenetic tree demonstrated that the subfamily Oestrinae does not exhibit monophyly. Additionally, it revealed that C. titillator is a sister group to the subfamily Gasterophilinae. Despite the scarcity of data available for the 28S rRNA gene, the phylogenetic analysis utilizing 28S rRNA revealed one distinct lineage for the Egyptian camel nasal bot fly.ConclusionsMolecular phylogenetic analysis was conducted using molecular markers of distinct origins (both mitochondrial and nuclear) to elucidate the evolutionary relationships within the family Oestridae. This analysis is particularly significant following the inclusion of C. titillator, a first-time discovery in Egypt.

Human fascioliasis emergence in southern Asia: Complete nuclear rDNA spacer and mtDNA gene sequences prove Indian patient infection related to fluke hybridization in northeastern India and Bangladesh

Fascioliasis is a snail-borne zoonotic disease with impact on the development of human subjects and communities. It is caused by two liver-infecting fasciolid trematode species, the globally-distributed Fasciola hepatica and the Africa/Asia-restricted but more pathogenic, larger F. gigantica. Fasciola gigantica is the cause of endemicity in livestock throughout the warm lowlands from Pakistan to southeastern Asia since old times. Human fascioliasis is emerging in this region at present, with an increase of patient reports. Complete sequences of rDNA ITS-1 and ITS-2 spacers and mtDNA nad1 and cox1 genes were obtained from fasciolid eggs found in the endoscopic bile aspirate from a patient of Arunachal Pradesh, northeastern India. Egg measurements, pronounced ITS heterozygosity, and pure F. gigantica mtDNA haplotypes demonstrate an infection by a recent F. gigantica-like hybrid. Sequence identities and similarities with the same DNA markers found in livestock from Bangladesh prove the human-infecting fasciolid to present identical ITSs and nad1 haplotypes and only one silent transversion in cox1 when compared to a widely-spread combined haplotype in animals. In northeastern India and Bangladesh, human fascioliasis emergence appears linked to increasing livestock prevalences due to: ruminant importation from other countries because of the increasing demand of rapidly growing human populations; numerous livestock movements, including transborder corridors, due to the uncontrolled small-scale household farming practices; and man-made introduction of F. hepatica with imported livestock into an area originally endemic for F. gigantica leading to frequent hybridization. Sequences, phylogenetic trees, and networks indicate that the origins of intermediate/hybrid fasciolids and factors underlying human infection risk differ in eastern and western South Asia. The emergence scenario in southern China and Vietnam resembles the aforementioned of northeastern India and Bangladesh, whereas in Pakistan it is linked to increasing monsoon rainfall within climate change combined with an impact of an extensive irrigation system. Past human-guided movements of pack animals along the western Grand Trunk Road and the eastern Tea-Horse Road explain the F. gigantica mtDNA results obtained. Physicians should be aware about these emerging scenarios, clinical pictures, diagnostic techniques and treatment. Government authorities must appropriately warn health professionals, ensure drug availability and improve livestock control.

Influence of COVID-19 on the emergence of stone-tool use behavior in a population of common long-tailed macaques (Macaca fascicularis fascicularis) in Thailand.

Stone tool use is a rare behavior across nonhuman primates. Here we report the first population of common long-tailed macaques (Macaca fascicularis fascicularis) who customarily used stone tools to open rock oysters (Saccostrea forskali) on a small island along the Thai Gulf in Koh Ped (KPE), eastern Thailand. We observed this population several times during the past 10 years, but no stone-tool use behavior was observed until our survey during the coronavirus disease 2019 (COVID-19) pandemic in July 2022. KPE is located in Pattaya City, a hotspot for tourism in Thailand. Tourists in this area frequently provided large amounts of food for the monkeys on KPE. During the COVID-19 curfew, however, tourists were not allowed to access the island, and monkeys began to face food scarcity. During this time, we observed stone-tool use behavior for the first time on KPE. Based on our observations, the first tool manipulation was similar to stone throwing (a known precursor of stone tool use). From our observations in March 2023, we found 17 subadult/adult animals performing the behavior, 15 of 17 were males and mostly solitary while performing the behavior. The M. f. fascicularis subspecies was confirmed by distribution, morphological characteristics, and mtDNA and SRY gene sequences. Taken together, we proposed that the stone tool use behavior in the KPE common long-tailed macaques emerged due to the COVID-19 food scarcity. Since traveling is no longer restricted many tourists have started coming back to the island, and there is a high risk for this stone tool-use behavior to disappear within this population of long-tailed macaques.

Plasma mitochondrial DNA is elevated in maternal serum at first trimester and may serve as a biomarker for prediction of gestational diabetes mellitus.

We evaluated whether an abundance of first-trimester plasma mitochondrial DNA (mtDNA) fragments could predict the risk for the development of gestational diabetes mellitus (GDM) by the late second or early third trimester. It was a prospective study wherein we enrolled 150 women in their first trimester of gestation. Oral glucose tolerance test (OGTT) was administered both in the first and second trimesters to diagnose GDM. Among our cohort, 23 women were diagnosed with GDM in the first trimester and excluded from the study. Of the remaining 127, 29 women were diagnosed with GDM in the second trimester, and 98 women who did not develop GDM served as controls. We amplified blood drawn from each participant during the first trimester for three distinct mtDNA gene sequences: COX, ND4, and D-loop. An abundance of each mtDNA sequence, estimated by the ΔCt method between mtDNA and 18S rRNA, correlated with GDM occurrence in the late second or early third trimester. There was a significant difference in ΔCt COX between controls and those with GDM occurrence in the second trimester (p = .006). These levels were not associated with age or fasting plasma glucose levels in the first trimester. ΔCt COX could predict GDM with a sensitivity of 90% and a specificity of 40%. Though ΔCt ND4 was higher in the GDM-positive group, the levels did not reach statistical significance. ΔCt D-loop was similar in GDM-positive cases and controls who did not develop GDM during pregnancy. These results were in plasma samples collected 3 to 4 months before overt hyperglycemia diagnosis suggestive of GDM. The abundance of plasma mtDNA fragments represents a promising cost-effective, convenient early-stage biomarker for predicting GDM development. Importantly, it can be administered irrespective of the fasting status of the subject. Further assessment of the predictive capacity of these biomarkers within large, diverse populations is needed for effective clinical utility.

A new species of the genus Hemiphyllodactylus Bleeker, 1860 (Squamata: Gekkonidae) from a lowland monsoon forest in Southern Vietnam.

An integrative taxonomic analysis delimits and diagnoses the new species Hemiphyllodactylus cattien sp. nov. from Cat Tien National Park, Dong Nai Province, southern Vietnam-the eighth species of Hemiphyllodactylus to be described from Vietnam. The new species is recovered as the sister species to a clade comprised of H. indosobrinus from Laos and H. flaviventris from Thailand based on phylogenetic analyses using ND2 mtDNA gene sequences. Unlike the vast majority of Hemiphyllodactylus which are site-restricted upland endemics, Hemiphyllodactylus cattien sp. nov. is a lowland species collected from one of the most well-surveyed regions in Vietnam. Its discovery further emphasizes the need for continued re-sampling of survey sites already considered to be well-explored.

The Phylogeny of Bornean Swamp Buffalo (Bubalus bubalis) Analysis Based on D-loop Mitochondrial DNA Sequence Variation

Swamp buffalo ( Bubalus bubalis ) is one of the Indonesian germplasm that adapts well to Borneo Island. This study aimed to determine the genetic diversity, phylogenetic tree, and phylogeographic structure of Bornean swamp buffalo based on control region (D-loop) mitochondrial DNA sequences. A total of 120 animals were collected from three populations in Indonesia, i.e., the North, East, and South of Kalimantan Provinces. Along 1140 bp of D-loop mtDNA gene of Bornean swamp buffaloes was amplified with a design primer of F: 5'-CAA CAC CCA AAG CTG AAG TT-3’ and R: 5'-CGC TCC TCT TAG TCT CGT TG-3’. Therefore, the forward and reverse sequencing was performed to visualize the full length of D-loop mtDNA gene sequence (1140 bp). Results showed that a total of 47 haplotypes were detected in the animal study, with haplotype and nucleotide diversities of 0.936 and 0.005, respectively. Consequently, two haplogroups were observed in the animal study, i.e., Haplogroup A (North) and Haplogroup B (East and South). Based on structure analysis, Bornean swamp buffaloes were comparable to filing swamp buffaloes of China based on structure. According to the analysis of molecular (AMOVA), the geographical component contributed over 56.44% of the total mtDNA sequence variations. In conclusion, it was discovered that the haplogroups of buffalo from the East and South populations were identical.

Oswaldocruzia ukrainae (Nematoda: Molineidae)-A Parasite of European Green Toad Bufotes viridis: Morphological and Molecular Data.

Nematodes of the genus Oswaldocruzia are common parasites of the small intestine of amphibians and reptiles. Our recent molecular analysis of Oswaldocruzia nematodes revealed that only Oswaldocruzia filiformis, which possesses high morphological variability, parasitizes amphibians and reptiles in European Russia. Here we present the study of Oswaldocruzia nematodes from the European green toad Bufotes viridis (Anura, Bufonidae) collected at different localities of the Middle Volga region in 2018-2022. We analyzed the morphological characteristics of the Oswaldocruzia spp. taxonomy together with novel molecular phylogenetic data. The data on phylogenetic analysis (based on partial CoxI mtDNA gene sequences) showed that Bufotes viridis is parasitized by two Oswaldocruzia species, the host-specific parasite Oswaldocruzia ukrainae and species generalist Oswaldocruzia filiformis. Broad morphological variability was revealed in O. ukrainae nematodes both from the same host specimen and from various toad individuals from different localities. Our results highlight the need for further biodiversity research of morphologically similar Oswaldocruzia species from amphibians and reptiles in the Western Palearctic using molecular genetic methods.

Three New Species of Jainus (Monogenea: Dactylogyridae) Parasitizing Gills of Brazilian Freshwater Fishes Supported by Morphological and Molecular Data

The present study describes three new species of monogeneans parasitizing the gills of anostomid fishes from the Upper Paraná River basin, southeastern Brazil: Jainus beccus n. sp. and Jainus radixelongatus n. sp. on Leporinus friderici, Leporinus octofasciatus, Leporinus striatus, and Megaleporinus elongatus; and Jainus ornatus n. sp. on L. friderici. The new species differ from other congeners by the morphology of the accessory piece. There is a semicircular distal portion resembling a “bird’s beak” in Jainus beccus n. sp. It composed of two subunits—one ventral and more sclerotized, sickle-shaped, and another dorsal with three projections—in Jainus radixelongatus n. sp. There are two elongated and sclerotized subunits, both of which have a sickle-shaped distal portion, in Jainus ornatus n. sp. Supplementary observations not reported in the original descriptions of the type-species Jainus piava Karling, Bellay, Takemoto & Pavanelli, 2011 are proposed, as follows: the presence of a thin and delicate ventral bar, which can vary greatly in shape; an accessory piece not articulated with the MCO’s base. This paper provides the first phylogenetic study based on LSU rDNA and COI mtDNA gene sequences for Jainus, improving and clarifying the understanding of host–parasite relationships in neotropical characiforms.

The North Asian Genus Kolhymamnicola Starobogatov and Budnikova 1976 (Gastropoda: Amnicolidae), Its Extended Diagnosis, Distribution, and Taxonomic Relationships

The taxonomic position and phylogenetic affinities of the endemic North Asian genus Kolhymamnicola Starobogatov and Budnikova, 1976 (Gastropoda: Amnicolidae) remain unknown. To resolve this, we studied key morpho-anatomical characteristics of Kolhymamnicola snails and performed a molecular phylogenetic analysis based on sequences of COI mtDNA, 16S rRNA, and 18S rRNA genes. In terms of protoconch microsculpture, operculum, radular teeth, and gill complex morphology, Kolhymamnicola snails do not differ significantly from the North American genera Amnicola Gould and Haldeman, 1840 and Taylorconcha Hershler et al., 1994, and the European genus Marstoniopsis van Regteren Altena 1936. The bifid penis found in Kolhymamnicola is similar to that in the genus Marstoniopsis. The female reproductive anatomy has some features shared by Kolhymamnicola and Taylorconcha (absence of bursa copulatrix, single seminal receptacle in rs2′ position, and ventral channel). The molecular analysis has revealed Taylorconcha as the closest relative to Kolhymamnicola; the COI-based genetic distance between them amounted to 0.113. We discuss the possible time of divergence of these two genera, as well as of European Marstoniopsis and the Baikal Lake endemic family Baicaliidae. The last common ancestor of these groups was widely distributed in Miocene–Pliocene in the Holarctic waterbodies. Recent Kolhymamnicola snails are distributed in Northern Asia, including lakes of the Baikal rift zone. We rank the Baicaliidae as a family rather than a subfamily of Amnicolidae based on their distinct, unique morpho-anatomical characteristics and highly supported separate position on the molecular tree. The tribe Erhaiini Davis and Kuo, 1985 is elevated to the rank of the family, with 3–4 recent genera included. The family Palaeobaicaliidae Sitnikova et Vinarski fam. nov. is established to embrace the Cretaceous North Asian gastropods conchologically similar to the recent Baicaliidae and Pyrgulidae.

Two New Species of Leptobrachella from Northern Thailand (Amphibia, Anura, Megophryidae)

Phylogenetic analyses using mtDNA gene sequences revealed the presence of two genetic clades in the genus Leptobrachella (former Leptolalax) from northern Thailand. One formed a clade including L. ventripunctata and L. aerea, while another included L. bourrreti and L. laoi. The first clade contained L. cf. minima from Doi Suthep, while the second clade contained L. murphyi and two undescribed species, one from Doi Inthanon, at a high altitude of 2,300 m asl, and another from Mae Hong Son, at Myanmar border. These form a clade, that is sister to L. murphyi. Morphologically, the first species is generally similar to L. murphyi, while the second species resembles L. cf. minima. However, they are clearly distinguished from those described species by great genetic differences, and we describe them as L. korifi sp. nov. and L. sinorensis sp. nov., respectively.

Resurrection of Pareas yunnanensis (Vogt, 1922) with description of a new species of Pareas from Yunnan Province, China (Squamata, Pareidae)

Based on molecular and morphological data of four specimens of Pareas Wagler, 1830 collected from the type locality of P. yunnanensis (Vogt, 1922), along with examination of the type specimens of P. yunnanensis, we revalidate this poorly known, secretive species. Furthermore, based on molecular and morphological lines of evidence we also describe a new species of Pareas from Xishuangbanna Prefecture, Yunnan Province, China. Morphologically, the new species closely resembles its sister species P. nigriceps Guo & Deng, 2009. However, the new species is divergent from the latter in cytochrome b mtDNA gene sequences, and can be distinguished from all congeners by the following combination of morphological characteristics: single preocular, postocular fused with subocular, loreal not bordering orbit, vertebral scales enlarged, 3–5 rows of mid-dorsal scales keeled at the middle of the body, ventral scales 160–171; subcaudals 62–64, dorsal surface of head solid black or reddish-brown, dark nuchal band present, iris brownish-black or reddish-brown.

Phylogeography of the Maleo Senkawor (Macrocephalon maleo Sal. Muller 1846) Based on Cytochrome B Gene in Sulawesi And Their Sex Determination using Molecular Sexing

The Maleo Senkawor bird (Macrocephalon maleo) is endemic and widely distributed in Sulawesi. Maleo Senkawor bird is monomorphic so it is difficult to distinguish between male and female individuals. This study aimed to determine the kinship relationship among M. maleo in various geographic areas in Sulawesi through the use of the gene marker Cytochrome-b (Cyt-b), determine the sex ratio, and examine the reliability of the 2550F/2718R primer. Fifteen (feather and eggshell) samples from North Sulawesi, Southeast Sulawesi, and Central Sulawesi were collected. Total DNA isolation was performed using the Dneasy® Blood and Tissue kit (Cat. No. 69504) following a Qiagen protocol with modification. PCR amplification (35 cycles) used a forward primer MMCytb_F (5'- GAAAATCCCACCCCCTACTA-3'), a reverse primer MMCytb_R (5'- GTTGGCTACGAGGAGTCAGA-3') and a primer for sexing used 2550F/2718R. Analysis was performed on the length of 903 bp Cyt-b mtDNA gene sequences of M. maleo along with their amino acid sequences (301 AA) and Chromo Helicase DNA-Binding (CHD) gene sequences. Based on the genetic distance of the Kimura 2-Parameter model and p-distance, the interpopulation of M. maleo in Sulawesi (the population of Southeast Sulawesi was separated from the population of Central Sulawesi and North Sulawesi) was 0.002 (0.2%) – 0.003 (0.3%). Each population has formed a different haplotype. Primer 2550F/2718R was able to amplify the CHD gene and could distinguish the sex identification. Male individuals are more dominant than female individuals with a sex ratio of 6,5:1

Characterization of Hoplolaimus seinhorsti and Hoplolaimus pararobustus (Tylenchina: Hoplolaimidae) from banana, with phylogeny and species delineation in the genus Hoplolaimus.

The morphological and molecular characterisations of two lance nematode species isolated from the rhizosphere of banana, Hoplolaimus seinhorsti and H. pararobustus, are provided based on an integrative study that includes light and scanning electron microscopy, phylogenetic analysis and two tree-based molecular species delimitation methods (GMYC and bPTP). Nineteen new sequences were obtained, including 5 partial 18S rRNA, 6 D2-D3 of 28S rRNA, 1 ITS rRNA and 7 COI mtDNA (the first COI sequences of H. seinhorsti and H. pararobustus), and an updated morphological character comparison of 37 Hoplolaimus species is presented. The tree-based molecular species-delimitation approaches employed gave markedly differing results, and also showed remarkable discrepancies among the investigated genes, although the bPTP output was found to agree well with established morphological species delimitations. Both species-delimitation approaches did, however, provide the same output for the COI mtDNA sequences, and the COI mtDNA gene sequence was also found to correspond better to established morphological species. It is therefore recommended by this paper as representing the most suitable barcode marker for Hoplolaimus species identification. This integrative study also resulted in the corrective reassignment of 17 gene sequences that were previously unidentified or incorrectly classified, as well as concluding that H. pararobustus consists of two cryptic species.

Türkiye Podarcis siculus siculus (Rafinesque-Schmaltz, 1810) Populasyonlarının Kökenini Ortaya Çıkarmak için Tür Sınırlarının Belirlenmesi

It is possible that the origin of Podarcis sicurus, which was introduced to Turkey by humans, is based on two different source populations or that the samples introduced to different geographies may differ at the subspecies level. The rapid evolutionary processes observed in island species can similarly occur in introduced and isolated populations. In this study Tissues were taken from samples obtained from Zonguldak, Istanbul and Samsun. mtDNA gene sequences were obtained and combined with Podarcis sicurus samples from sequences downloaded from Genbank. Phylogenetic analyzes and species delimitation analyzes were executed. Accordingly results, the specimens obtained from Turkey were grouped into two different haplotypes, similar to their geographic isolation. It has been observed that these groups are different resource groups. Species delimination analyzes show that the differentiation of these haplotypes may be at the subspecies level.

Molecular evidence for introgressive hybridization in New Zealand masked gulls

We used mitochondrial DNA (mtDNA) gene sequences and nuclear microsatellite loci to investigate the extent and outcome of hybridization between the Black‐billed Gull Chroicocephalus bulleri and the Red‐billed Gull Chroicocephalus novaehollandiae scopulinus in New Zealand. Six of 26 sampled Black‐billed Gulls possessed mtDNA typical of Red‐billed Gulls, but allele frequencies at six polymorphic microsatellites provided little evidence of mixed ancestry expected in very recent hybrids. None of the Red‐billed Gulls sampled from different colonies possessed Black‐billed Gull mtDNA expected in the reciprocal cross, suggesting that hybridization in the two species typically occurs between female Red‐billed Gulls and Black‐billed Gull males. The lack of any hybrid signal in the nuclear loci indicates that there has been extensive backcrossing with Black‐billed Gulls, effectively diluting the Red‐billed Gull nuclear DNA contribution. Divergence of Red‐billed Gulls and Black‐billed Gulls occurred approximately 250 000 years ago, indicating that unsorted ancestral polymorphism is an unlikely alternative to hybridization. Comparing demographic models within an approximate Bayesian computation (ABC) framework, we confirm that the observed patterns cannot result from incomplete lineage sorting. Using an ABC random forest approach, we determined that the most likely model explaining the data is a recent introgression scenario, whereby unidirectional gene flow is re‐established following a period of strict isolation. The ability of Black‐billed and Red‐billed Gulls to successfully interbreed shows that despite significant differentiation (FST > 0.3), there has been insufficient time for the two species to develop complete reproductive isolation. The apparent one‐way transfer of Red‐billed Gull mtDNA into Black‐billed Gulls and extensive backcrossing argues against cytoplasmic–nuclear genome incompatibilities between the two species. We hypothesize that the specific mate recognition system cued on colours of soft parts normally functions to prevent hybridization, but that it can break down under demographic conditions where there is a shortage of available mates and a surplus of females in the Red‐billed Gull population. The high incidence of introgression in Black‐billed Gulls conflicts with field observations that interbreeding is extremely rare.

Enenterum kyphosi Yamaguti, 1970 and Enenterum petrae n. sp. (Digenea: Enenteridae) from kyphosid fishes (Centrarchiformes: Kyphosidae) collected in marine waters off eastern Australia.

Species of the digenean genus Enenterum Linton, 1910 (Lepocreadioidea: Enenteridae) are characterised primarily by their elaborate oral suckers, which are divided into varying numbers of anteriorly directed lobes, and their host-restriction to herbivorous marine fishes of the family Kyphosidae. We describe Enenterum petrae n. sp. from the brassy chub Kyphosus vaigiensis (Quoy Gaimard) collected off Lizard Island, Great Barrier Reef, Queensland, Australia. Enenterum petrae n. sp. is readily differentiated from congeners by its unique oral sucker morphology, in having a minute pharynx, and the combination of a genital cap and accessory sucker. We also provide the first record of Enenterum kyphosi Yamaguti, 1970 from Australia based on material obtained from the blue sea chub Kyphosus cinerascens (Forsskl) collected off Lizard Island and North Stradbroke Island, Queensland. Morphologically, our specimens of E. kyphosi agree closely with descriptions of this species from Hawaii and South Africa, and despite lack of molecular data from outside of Australian waters, we consider all three reports to represent a single, widespread species. The first ITS2 and COI mtDNA gene sequences for species of Enenterum are provided and molecular phylogenetic analyses of 28S rDNA gene sequences place these species in a strongly-supported clade with the type-species of the genus, Enenterum aureum Linton, 1910. The oral suckers of both E. kyphosi and E. petrae n. sp. can be interpreted as having varying numbers of lobes depending on the particular specimen and how the division between lobes is defined. Scanning electron microscopical images improves understanding of the morphology of the enenterid oral sucker, and permits speculation regarding the evolutionary history leading to its specialisation in this lineage.