Once widespread in their homelands, the Anatolian mouflon (Ovis gmelini anatolica) and the Cyprian mouflon (Ovis gmelini ophion) were driven to near extinction during the 20th century and are currently listed as endangered populations by the International Union for Conservation of Nature. While the exact origins of these lineages remain unclear, they have been suggested to be close relatives of domestic sheep or remnants of proto-domestic sheep. Here, we study whole genome sequences of n = 5 Anatolian mouflons and n = 10 Cyprian mouflons in terms of population history and diversity, comparing them with eight other extant sheep lineages. We find reciprocal genetic affinity between Anatolian and Cyprian mouflons and domestic sheep, higher than all other studied wild sheep genomes, including the Iranian mouflon (O. gmelini). Studying diversity indices, we detect a considerable load of short runs of homozygosity blocks (<2 Mb) in both Anatolian and Cyprian mouflons, reflecting small effective population size (N-e). Meanwhile, N-e and mutation load estimates are lower in Cyprian compared with Anatolian mouflons, suggesting the purging of recessive deleterious variants in Cyprian sheep under a small long-term N-e, possibly attributable to founder effects, island isolation, introgression from domestic lineages, or differences in their bottleneck dynamics. Expanding our analyses to worldwide wild and feral Ovis genomes, we observe varying viability metrics among different lineages and a limited consistency between viability metrics and International Union for Conservation of Nature conservation status. Factors such as recent inbreeding, introgression, and unique population dynamics may have contributed to the observed disparities.

We present palaeogenomes of three morphologically unidentified Anatolian equids dating to the first millennium BCE, sequenced to a coverage of 0.6-6.4x. Mitochondrial DNA haplotypes of the Anatolian individuals clustered with those of Equus hydruntinus (or Equus hemionus hydruntinus), the extinct European wild ass, secular name 'hydruntine'. Further, the Anatolian wild ass whole genome profiles fell outside the genomic diversity of other extant and past Asiatic wild ass (E. hemionus) lineages. These observations suggest that the three Anatolian wild asses represent hydruntines, making them the latest recorded survivors of this lineage, about a millennium later than the latest observations in the zooarchaeological record. Our mitogenomic and genomic analyses indicate that E. h. hydruntinus was a clade belonging to ancient and present-day E. hemionus lineages that radiated possibly between 0.6 and 0.8 Mya. We also find evidence consistent with recent gene flow between hydruntines and Middle Eastern wild asses. Analyses of genome-wide heterozygosity and runs of homozygosity suggest that the Anatolian wild ass population may have lost genetic diversity by the mid-first millennium BCE, a possible sign of its eventual demise.

A major challenge in zooarchaeology is to morphologically distinguish closely related species’ remains, especially using small bone fragments. Shotgun sequencing aDNA from archeological remains and comparative alignment to the candidate species’ reference genomes will only apply when reference nuclear genomes of comparable quality are available, and may still fail when coverages are low. Here, we propose an alternative method, MTaxi, that uses highly accessible mitochondrial DNA (mtDNA) to distinguish between pairs of closely related species from ancient DNA sequences. MTaxi utilises mtDNA transversion-type substitutions between pairs of candidate species, assigns reads to either species, and performs a binomial test to determine the sample taxon. We tested MTaxi on sheep/goat and horse/donkey data, between which zooarchaeological classification can be challenging in ways that epitomise our case. The method performed efficiently on simulated ancient genomes down to 0.3x mitochondrial coverage for both sheep/goat and horse/donkey, with no false positives. Trials on n=18 ancient sheep/goat samples and n=10 horse/donkey samples of known species identity also yielded 100% accuracy. Overall, MTaxi provides a straightforward approach to classify closely related species that are difficult to distinguish through zooarchaeological methods using low coverage aDNA data, especially when similar quality reference genomes are unavailable. MTaxi is freely available at https://github.com/goztag/MTaxi.

The social organization of the first fully sedentary societies that emerged during the Neolithic period in Southwest Asia remains enigmatic,(1) mainly because material culture studies provide limited insight into this issue. However, because Neolithic Anatolian communities often buried their dead beneath domestic buildings,(2) household composition and social structure can be studied through these human remains. Here, we describe genetic relatedness among co-burials associated with domestic buildings in Neolithic Anatolia using 59 ancient genomes, including 22 new genomes from Asxikli Hoyuk and Catalhoyuk. We infer pedigree relationships by simultaneously analyzing multiple types of information, including autosomal and X chromosome kinship coefficients, maternal markers, and radiocarbon dating. In two early Neolithic villages dating to the 9th and 8th millennia BCE, Asxikli Hoyuk and Boncuklu, we discover that siblings and parent-offspring pairings were frequent within domestic structures, which provides the first direct indication of close genetic relationships among co-burials. In contrast, in the 7th millennium BCE sites of Catalhoyuk and Barcin, where we study subadults interred within and around houses, we find close genetic relatives to be rare. Hence, genetic relatedness may not have played a major role in the choice of burial location at these latter two sites, at least for subadults. This supports the hypothesis that in Catalhoyuk,(3-5) and possibly in some other Neolithic communities, domestic structures may have served as burial location for social units incorporating biologically unrelated individuals. Our results underscore the diversity of kin structures in Neolithic communities during this important phase of sociocultural development.

Sheep were among the first domesticated animals, but their demographic history is little understood. Here we analyzed nuclear polymorphism and mitochondrial data (mtDNA) from ancient central and west Anatolian sheep dating from Epipaleolithic to late Neolithic, comparatively with modern-day breeds and central Asian Neolithic/Bronze Age sheep (OBI). Analyzing ancient nuclear data, we found that Anatolian Neolithic sheep (ANS) are genetically closest to present-day European breeds relative to Asian breeds, a conclusion supported by mtDNA haplogroup frequencies. In contrast, OBI showed higher genetic affinity to present-day Asian breeds. These results suggest that the east-west genetic structure observed in present-day breeds had already emerged by 6000 BCE, hinting at multiple sheep domestication episodes or early wild introgression in southwest Asia. Furthermore, we found that ANS are genetically distinct from all modern breeds. Our results suggest that European and Anatolian domestic sheep gene pools have been strongly remolded since the Neolithic.

The genetic diversity of the Mediterranean swordfish (Xiphias gladius Linneus) has not been explored extensively at its easternmost range so far. In this study, modern X. gladius samples from the eastern part of the Mediterranean basin, north of the Aegean Sea (Aegean-2013, n = 26) and the Mediterranean coast of Turkey (N.Levantine-2013, n = 42) were studied genetically, along with ancient samples from Yenikapi excavation (n = 6). Partial mitochondrial DNA control region sequences (entire sequences, clade I and clade II) were evaluated spatially and temporally together with previously published sequences (Alvarado Bremer et al., Molecular Phylogenetics and Evolution, 2005, 36, 169-187; Vinas et al., ICES Journal of Marine Science, 2010, 67, 1222-1229; Righi et al., Diversity, 2020, 12, 170) from the rest of the Mediterranean Sea. Pair-wise F-ST and pair-wise AMOVA tests showed that, in general, groups of eastern populations and western Mediterranean populations have not genetically differed from each other significantly nearly in the past 20 years. Therefore, the results direct reconsideration of previous descriptions of population sub-structure within the Mediterranean and support high gene flow throughout the region. On the contrary, the results of this study confirmed the existence of genetic diversity differences between western and eastern Mediterranean, with eastern being low. One-tailed permutation tests revealed that theta, which is directly proportional to long-term female effective population size (Ne), decreased significantly (P < 0.05) in both regions over the past two decades. On the Turkish coasts, theta is not significantly different from that of the nearly contemporary eastern Mediterranean population. Nonetheless, theta of the ancient sample was consistently and significantly (P < 0.001) higher than those of the eastern and western Mediterranean populations in clade I and clade II. Furthermore, it contains two mitochondrial haplotypes that are not observed in modern samples, suggesting that the Ne of X. gladius in the eastern was high in Byzantium times. Eight microsatellite loci were also genotyped in modern samples. The microsatellite-based present Ne estimate of the pooled Aegean-2013 and N.Levantine-2013 populations was lower than 1000 according to the upper limit of 95% c.i. and possibly even lower than 100 according to the mean of posterior distribution of the present Ne estimate calculated by the software package MSVAR. These alarming genetic signals for the sustainability of X. gladius on the coasts of Turkey are in agreement with the nearly collapsing X. gladius fisheries as depicted also in the fisheries statistics. Overall, congruent with the previous studies, the data presented here show that sustainability of the X. gladius population in Mediterranean is under major threat. Therefore, X. gladius around the Turkish coasts need an immediate stringent action and management plan.

Objective: The genetic diversity parameters for gazelle populations sampled in Turkey were estimated to assess the effects of captive breeding on the populations’ gene pools and effective population sizes. Materials and Methods: Four individuals from a recently discovered Gazella gazella population in Hatay and two captive gazelle populations were sampled (the Kizilkuyu State Farm (n=48) and the Erikce State Farm (n=25)) and analyzed using nuclear DNA, mtDNA and Y-chromosome markers. Results: The mtDNA cyt-b partial sequence analysis assigned the Erikce and Kizilkuyu samples to Gazella marica. The structure analysis differentiated significantly between them, and revealed samples originating from wild population. Both, the Y-chromosome INRA126 locus sequences of Gazella gazella and Gazella marica males and the mtDNA partial cyt-b region RFLP analysis from all the samples distinguished the two gazelle species from each other. Based on microsatellites, the estimated effective population sizes were 9.7, 8.9 and 6.4 for the Kizilkuyu, Erikce and Hatay populations, respectively. When the Kizilkuyu and Erikce populations (where severe inbreeding depressions seems to be occurring already) were pooled, the estimated Ne was 24.5. All these estimates were too small for the sustainability of either individual or pooled populations in the wild or even in captivity. Conclusion: The markers used in the study provided information on two of the gazelle species (Gazella marica, and Gazella gazella): their species identity, degree of divergences, effective population sizes and the presence of admixture within the populations. These results turned out to be invaluable in terms of their contribution to future studies for the conservation of these species.

In this study, 182 male animals from 12 native sheep breeds, as well as Karacabey Merino and Karagül breeds of Anatolia, wild sheep Anatolian Mouflon (Ovis gmelinii anatolica) were used as the study material. Based on SRY and SRYM18 regions on the Y-chromosome, haplotypes of the populations were analyzed using DNA sequence analyses. The SRY region, A-oY1 allele was observed in all of the individuals studied. On the other hand, four different alleles corresponding to four Y-chromosome haplotypes were detected at the SRYM18 microsatellite region. Among native Anatolian breeds (n=143), H6 haplotype (80.41%), H4 haplotype (9.09%), H8 haplotype (8.40%) and H12 haplotype (2.1%) were identified. H6 haplotype was observed in all 16 individuals of Ovis gmelinii anatolica. Pairwise FST values based on haplotype frequencies were calculated for domestic sheep, and the highest FST value was observed between Karagül and Kıvırcık along with Karagül and Ovis gmelinii anatolica with pairwise FST value of 0.43202 (P<0.01). Y chromosome polymorphism of sheep from Turkey were examined comparatively with the accumulated data in the literature. Out of seven haplotypes (H4, H5, H6, H7, H8, H12, H19) observed in Europe and Asia, 4 haplotypes (H4, H6, H8 and H12) were observed in Anatolia. H12 was a private haplotype of Sakız, H6 seems to be the predominant haplotype of domestic sheep (79.51%) as well as being the only haplotype observed in Ovis gmelini anatolica. H4 haplotype seemed to be associated with fat tailed sheep migrating to Turkey, entering from south east of Turkey, which may be related with the arrival of nomadic Turks.

OBJECTIVES:North Mesopotamia has witnessed dramatic social change during the Holocene, but the impact of these events on its demographic history is poorly understood. Here, we study this question by analysing genetic data from the recently excavated Late Iron Age settlement of Çemialo Sırtı in Batman, southeast Turkey. Archaeological and radiocarbon evidence indicate that the site was inhabited during the second and first millennia BCE. Çemialo Sırtı reveals nomadic items of the Early Iron Age, as well as items associated with the Late Achaemenid and subsequent Hellenistic Periods. We compare Çemialo Sırtı mitochondrial DNA profiles with earlier and later populations from west Eurasia to describe genetic continuity patterns in the region.MATERIALS AND METHODS:A total of 16 Çemialo Sırtı individuals' remains were studied. PCR and Sanger sequencing were used to obtain mitochondrial DNA HVRI-HVRII sequences. We studied haplotype diversity and pairwise genetic distances using FST , comparing the Çemialo Sırtı population with ancient and modern-day populations from west Eurasia. Coalescent simulations were carried out to test continuity for specific population comparisons.RESULTS:Mitochondrial DNA (mtDNA) haplotypes from 12 Çemialo Sırtı individuals reveal high haplotype diversity in this population, conspicuously higher than early Holocene west Eurasian populations, which supports the notion of increasing population admixture in west Eurasia through the Holocene. In its mtDNA composition, Çemialo Sırtı shows highest affinity to Neolithic north Syria and Neolithic Anatolia among ancient populations studied, and to modern-day southwest Asian populations. Based on population genetic simulations we cannot reject continuity between Neolithic and Iron Age, or between Iron Age and present-day populations of the region.DISCUSSION:Despite the region's complex sociopolitical history and indication for increased genetic diversity over time, we find no evidence for sharp shifts in north Mesopotamian maternal genetic composition within the last 10,000 years.

The Neolithic transition in west Eurasia occurred in two main steps: the gradual development of sedentism and plant cultivation in the Near East and the subsequent spread of Neolithic cultures into the Aegean and across Europe after 7000 cal BCE. Here, we use published ancient genomes to investigate gene flow events in west Eurasia during the Neolithic transition. We confirm that the Early Neolithic central Anatolians in the ninth millennium BCE were probably descendants of local hunter-gatherers, rather than immigrants from the Levant or Iran. We further study the emergence of post-7000 cal BCE north Aegean Neolithic communities. Although Aegean farmers have frequently been assumed to be colonists originating from either central Anatolia or from the Levant, our findings raise alternative possibilities: north Aegean Neolithic populations may have been the product of multiple westward migrations, including south Anatolian emigrants, or they may have been descendants of local Aegean Mesolithic groups who adopted farming. These scenarios are consistent with the diversity of material cultures among Aegean Neolithic communities and the inheritance of local forager know-how. The demographic and cultural dynamics behind the earliest spread of Neolithic culture in the Aegean could therefore be distinct from the subsequent Neolithization of mainland Europe.

The archaeological documentation of the development of sedentary farming societies in Anatolia is not yet mirrored by a genetic understanding of the human populations involved, in contrast to the spread of farming in Europe [1-3]. Sedentary farming communities emerged in parts of the Fertile Crescent during the tenth millennium and early ninth millennium calibrated (cal) BC and had appeared in central Anatolia by 8300 cal BC [4]. Farming spread into west Anatolia by the early seventh millennium cal BC and quasi-synchronously into Europe, although the timing and process of this movement remain unclear. Using genome sequence data that we generated from nine central Anatolian Neolithic individuals, we studied the transition period from early Aceramic (Pre-Pottery) to the later Pottery Neolithic, when farming expanded west of the Fertile Crescent. We find that genetic diversity in the earliest farmers was conspicuously low, on a par with European foraging groups. With the advent of the Pottery Neolithic, genetic variation within societies reached levels later found in early European farmers. Our results confirm that the earliest Neolithic central Anatolians belonged to the same gene pool as the first Neolithic migrants spreading into Europe. Further, genetic affinities between later Anatolian farmers and fourth to third millennium BC Chalcolithic south Europeans suggest an additional wave of Anatolian migrants, after the initial Neolithic spread but before the Yamnaya-related migrations. We propose that the earliest farming societies demographically resembled foragers and that only after regional gene flow and rising heterogeneity did the farming population expansions into Europe occur.

Archaeogenomic analysis of ancient Anatolians : first genetic indication for Neolithic cultural diffusion in the Near East

The aim of this study was to determine the genetic diversity of 4 native Turkish cattle breeds, based on the growth hormone (GH), prolactin (PRL), and diacylglycerol acyltransferase (DGAT1) genes. In order to study the polymorphisms in these genes, the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method was performed. A 329-bp fragment and a 223-bp fragment of GH, a 156-bp fragment of PRL, and a 411-bp fragment of DGAT1, thus 4 loci of 3 genes, were amplified via PCR. These fragments were then restricted with the enzymes MspI, Alu1, RsaI, and CfrI, respectively. In this study, 2 types of alleles, (+) and (-) for the GH-MspI, L and V for the GH-Alu1, A and B for the PRL, and K and A for the DGAT1 loci, were observed. The results of the present study will contribute to the polymorphism data on the world's cattle breeds. Furthermore, the above-mentioned allele frequencies of Turkish native breeds are evaluated in relation to their genetic relatedness, and to infer their milk production properties on the basis of the available literature. In turn, these results can be utilized for future breeding programs of dairy cattle in Turkey.

In this study, twenty microsatellite loci were used to define genetic diversity among 56 water buffalo samples. Their somatic tissues (the skin and cartilage tissues from ears) and DNAs were deposited in Turkish Gene Banks. Samples of healthy animals were collected from three different geographic regions: Northern Turkey, North-Western Turkey and Eastern Thrace. Three (loci CSSM57, ETH3) to ten (locus CSSM47) different alleles were identified per micro-satellite locus in a total of 103 alleles. PIC values for the micro-satellite loci analysed ranged from 0.14 (CSSM32) to 0.82 (CSSM47) with a mean of 0.4945. In all of the populations for each of the loci, the observed heterozygosities (H-o) were greater than the expected heterozygosities (H-E), indicating that populations suffered from bottleneck. H-E per population ranged between 0.5359 in the Black Sea Region and 0.5208 in the Aegean-South Marmara Region. Within the population, inbreeding estimates (F-IS) was positive in only four of the 20 loci analysed. Individuals of the different geographic populations did not cluster on the neighbour joining tree which was constructed on the basis of allele sharing distances. Population differentiation was further visualized by Factorial Correspondence Analysis and determined by the pairwise estimations of fixation index (F-ST) and Nei's standard genetic distance (D-a). The results revealed that populations have inertia (as depicted by Factorial Correspondence Analysis), they are differentiated significantly but little (by pairwise F-ST values) and the least genetic distance is between Black Sea Region and Thrace Region. Comparative studies indicated that the genetic diversity of water buffalo harboured in the Gene Banks of Turkey is at the lower end of the diversity spectrum. This study thus highlights the usefulness of heterologous bovine microsatellite markers to assess the genetic variability in Anatolian water buffalo breeds. Furthermore, the results can be utilized for future breeding strategies and conservation.

In this study, the genetic diversity of three milk protein genes namely beta-casein, kappacasein and beta-lactoglobulin was estimated in Turkish cattle breeds. Based on these genes, breeds in Turkey have been grouped as: 1) Eastern Anatolian Red, Anatolian Black and Southern Anatolian Red and 2) Turkish Grey, Turkish Holstein and Holstein Candidate Bulls. B alleles of the three studied genes, which were reported to be positively related with cheese yield and quality, seemed to be low-intermediate for beta-casein and kappa-casein but relatively high for beta-lactoglobulin in the first group of Turkish breeds compared to other breeds of the world. The kappa-casein E allele, which has a negative effect on cheese quality, is absent in Turkish cattle breeds, except in Holstein Candidate Bulls. Therefore, the results suggest that milk of Turkish native breeds is suitable for cheese making. Based on observations of the Turkish breeds, some suggestions were made regarding breeding practices in Turkey.

Two anchovy forms identified from their external morphology and designated as ‘Silver’ and ‘Blue’ anchovies were captured in Mersin Bay (Eastern Mediterranean Sea). Silver anchovy seemed to inhabit shallower waters (9–77m) than Blue anchovy (majority at depths of 69–111m). Significant differences in body shape, which was analyzed through geometric morphometrics, and in otolith shape, examined using Fourier analysis, separated the two forms. Length polymorphisms at two intron and 9 microsatellite loci showed strong genetic differences between the two forms. Mersin-Bay Blue anchovy were genetically related to Engraulis encrasicolus from the Western Mediterranean Sea. Mersin-Bay Silver anchovy possessed an original genetic composition that distinguished them from both E. encrasicolus and Engraulis albidus at the two intron loci, while presenting affinities to E. albidus based on microsatellite markers. Mersin-Bay Silver anchovy were assigned to the latter species, of which they are considered as a geographically isolated population. This is the first record of E. albidus in the Levantine basin and confirmation of its presence in the Eastern Mediterranean Sea. These results have implications for the management of anchovy populations and their habitats.