MANUSCRIT ACCEPTAT MANUSCRIT ACCEPTAT Detecting the T1 cattle haplogroup in the Iberian Peninsula from Neolithic to medieval times: new clues to continuous cattle migration through time Lídia Colominas ; Ceiridwen J. Edward ; Albano Beja-Pereira ; Jean-Denis Vigne ; Raquel M.Silva ; Pere Castanyer ; Joaquim Tremoleda ; Maria Saña Seguí ; Manuel Pérez-Ripoll ; Felix Goyache ; Christopher J. Howe ; Graeme Barker ; Mim A. Bower Revista Journal of Archaeological Science. Volume 59, July 2015, Pages 110-117 DOI https://doi.org/10.1016/j.jas.2015.04.014 Disponible 06/05/2015 en línia Data de publicació 07/2015 Per citar aquest document: Lídia Colominas, Ceiridwen J. Edwards, Albano Beja-Pereira, Jean-Denis Vigne, Raquel M. Silva, Pere Castanyer, Joaquim Tremoleda, Maria Saña Seguí, Manuel Pérez-Ripoll, Felix Goyache, Christopher J. Howe, Graeme Barker, Mim A. Bower, Detecting the T1 cattle haplogroup in the Iberian Peninsula from Neolithic to medieval times: new clues to continuous cattle migration through time, In Journal of Archaeological Science, Volume 59, 2015, Pages 110-117, ISSN 0305-4403, https://doi.org/10.1016/j.jas.2015.04.014. Aquest arxiu PDF conté el manuscrit acceptat per a la seva publicació. 1 MANUSCRIT ACCEPTAT Abstract The spread of domestic animals through time is one of the topics studied by archaeologists to assess human trade and migration. Here we present mitochondrial analysis of 42 archaeological cattle (Bos taurus) bone samples, from 16 different sites in the Iberian Peninsula and covering a broad timeframe (from the early Neolithic to the Middle Ages), to provide evidence about the origin and dispersion of the T1 cattle haplogroup in relation to human contacts and movements. The presence of the T1 haplotype in one sample from an early Neolithic site close to the Mediterranean coast of Iberia, and its continuing presence in the Peninsula during Roman and Medieval times, clearly demonstrates that T1 was not solely a Muslim or later introduction. Rather, our molecular data show evidence for a pioneer coastal colonisation of the Iberian Peninsula from the Mediterranean basin, followed by possible further colonisation, leading to a continuity of diversity through time. Highlights • We report the earliest dated evidence of T1 mtDNA haplotypes in Neolithic Iberia. • There is continuity of T1 in Iberia from the Neolithic to medieval period. • Therefore, T1 was not solely a Muslim (711 AD) or later introduction. • Rather, T1 persisted from the initial spread of agriculture from the Near East. • This was then followed by possible further colonisation from Europe and Africa. Keywords Ancient DNA ; Cattle moviments ; Iberian Peninsula ; Mitochondrial haplotypes ; Mediterranean routes 1. Introduction The origin and diversification of domestic taurine cattle (Bos taurus) has been extensively investigated using mitochondrial (mt) DNA analyses of both modern and ancient samples. The majority of modern taurine mtDNA sequences in Europe fall into one of four phylogenetically distinct, yet closely related, star-like haplogroups, termed T, T1, T2 and T3. High diversity has been documented in Anatolia and the Middle East for both modern (Troy et al., 2001) and Neolithic and Bronze Age (Bollongino et al., 2006, 2012) specimens. The diversity in Europe and in Africa are both subsets of that seen in the Near East, with the most frequently observed haplogroups being T3 and T1 respectively. These results have been interpreted as a scenario of a single Neolithic origin of all T haplogroups in the Near and Middle East and a subsequent spread towards Europe and Africa (Troy et al., 2001; Bollongino et al., 2006; Lenstra et al., 2014). However, it appears that the process of cattle domestication and diffusion was more complex than originally thought, with a fifth haplogroup, T5, and two novel haplotypes (Q and R), recently reported from a small number of extant Italian cattle (Achilli et al., 2008, 2009). In addition, evidence of T3 being predominant in Italian aurochs (Beja-Pereira et al., 2006; Mona et al., 2010; Lari et al., 2011) is suggestive that at least some 2 MANUSCRIT ACCEPTAT Mediterranean T3 maternal lineages may have a European, rather than a Southwest Asian, origin. Similarly to T3 in Europe, T1 is fixed in the majority of extant African cattle (Lenstra et al., 2014), but it also found at low numbers in some contemporary Iberian, Italian and Greek breeds (Bradley et al., 1996; Cymbron et al., 1999; BejaPereira et al., 2006; Dadi et al., 2009; Bonfiglio et al., 2012). Research supports a spread of domestic T1 cattle from the Near East towards southern Europe and the Mediterranean basin during the Neolithic (Troy et al., 2001; Bollongino et al., 2006; Achilli et al., 2008; Lenstra et al., 2014), with the possibility that hybridisation occurred between migrating domestic populations from the Near East and resident wild African populations (Achilli et al., 2008; Decker et al., 2014). Specifically for the Iberian Peninsula, different explanations for the presence of T1 mitotypes in Iberian cattle breeds have been hypothesised. There is evidence for a wholly or partially North African origin for domestic cattle in Iberia (Cymbron et al., 1999; Miretti et al., 2002; Anderung et al., 2005; Beja-Pereira et al., 2006; Ginja et al., 2010), and a number of time periods for these origins and subsequent migrations have been identified. Possible introgressions of North African cattle into Iberia have been attested to have occurred: (1) during the Muslim invasion and occupation in the 8th century AD (Cymbron et al., 1999, 2005; Beja-Pereira et al., 2006); (2) as a consequence of the colonial activities in the 18th century AD (Cymbron et al., 1999); or (3) due to recent gene flow from Africa derived in the 1960s and 1970s (Beja-Pereira et al., 2002, 2003). However, the T1 haplogroup has been observed in a single Bronze Age animal from the north of Iberia (Anderung et al., 2005), highlighting that T1 was not only a recent introduction into the area. Taking these considerations into account, the aim of the research reported in this paper was to understand the phylogeny of haplogroup T1 in the Iberian Peninsula and the processes that led to its current geographical and breed distribution. 2. Methods 2.1. Samples Archaeological bones were collected from 20 Bos taurus samples retrieved from archaeological sites located in different areas of the Iberian Peninsula (Table 1; Fig. 1). The time periods included were Neolithic, Bronze Age, Roman and Middle Ages. Table 1. Data of archaeological Spanish cattle used in this study. Table of archaeological cattle samples studied, with associated information. Lab Sampl code e code Site Area Period Date Fragment length NEOL0 Spain Cova de Neolith 5400 c Alicanti 240 bp 1 04 l'Or ic al. BC Haplogro up T3 Source this study 3 MANUSCRIT ACCEPTAT Lab Sampl code e code Site Area Period Date Fragment length Haplogro up Source Cueva NEOL0 Spain Atapuer Neolith 5400 c del 240 bp 2 05 ca ic al. BC Mirador T3 this study 5213– Neolith 5044 c 240 bp ic al. BC T1 this study T this study NEOL0 IBE6 3 BRON0 IBE4 1 La Draga Girona Gatas 1900– Bronze Almeria 1700 c 240 bp Age al. BC BRON0 MAD1 Portalón Burgos 2 7 Bronze 1800 c 240 bp Age al. BC T3 AY8471 99 BRON0 MAD2 Portalón Burgos 3 Bronze 1780 c 240 bp Age al. BC T3 AY8471 88 BRON0 MAD3 Portalón Burgos 4 Bronze 1780 c 240 bp Age al. BC T3 AY8471 89 BRON0 MAD5 Portalón Burgos 5 Bronze 1780 c 240 bp Age al. BC T3 AY8471 90 BRON0 MAD8 Portalón Burgos 6 Bronze 1780 c 240 bp Age al. BC T3 AY8471 92 BRON0 MAD9 Portalón Burgos 7 Bronze 1780 c 117 bp Age al. BC T/T3 AY8471 93 BRON0 MAD1 Portalón Burgos 8 0 Bronze 1780 c 240 bp Age al. BC T AY8471 94 BRON0 MAD1 Portalón Burgos 9 1 Bronze 1780 c 240 bp Age al. BC T3 AY8471 95 BRON1 MAD1 Portalón Burgos 0 4 Bronze 1780 c 240 bp Age al. BC T3 AY8471 96 BRON1 MAD1 Portalón Burgos 1 8 Bronze 1780 c 240 bp Age al. BC T3 AY8472 00 Valparai Cuenca so de Bronze 1780 c 240 bp T3 AY8472 BRON1 MAD5 4 MANUSCRIT ACCEPTAT Lab Sampl code e code 2 1 Site Area Abajo Period Age Date Fragment length Haplogro up al. BC Source 11 BRON1 MAD1 Portalón Burgos 3 6 Bronze 1740 c 240 bp Age al. BC T1 AY8471 98 BRON1 MAD6 Portalón Burgos 4 Bronze 1635 c 240 bp Age al. BC T3 AY8471 91 Girona Iron Age 5th– 3rd c. BC 119 bp T/T3 JX87655 6 Olivet ROMN0 MC37 d'en 1 Pujol Girona Roman 1st c. BC 119 bp T/T3 JX87655 7 Olivet ROMN0 MC38 d'en 2 Pujol Girona Roman 1st c. BC 119 bp T/T3 JX87655 8 ROMN0 MC1 3 Vilauba Girona Roman 1st c. AD 119 bp T/T3 JX87655 3 ROMN0 MC9 4 Baetulo Barcelon 2nd c. Roman a AD 119 bp T/T3 JX87655 4 1st–3rd 119 bp c. AD T/T3 JX87655 5 T1 this study IRON01 MC36 St. Julià Ramis Vila ROMN0 MC18 Ametller Girona 5 s ROMN0 IBE5 6 Son Fornés Roman 2nd Mallorca Roman BC-1st 240 bp c. AD ROMN0 Empúrie EM20 Girona 7 s Roman 3rd c. AD mosaic ? this study ROMN0 Empúrie EM21 Girona 8 s Roman 1st c. AD 67 bp T/T3 this study ROMN0 Empúrie EM22 Girona 9 s Roman 2nd c. AD 240 bp T3 this study 5 MANUSCRIT ACCEPTAT Lab Sampl code e code Date Fragment length Haplogro up Area Period ROMN1 Empúrie EM26 Girona 0 s Roman 3rd c. AD mosaic ? this study ROMN1 Empúrie EM27 Girona 1 s Roman 3rd c. AD 316 bp T3 this study ROMN1 Empúrie EM28 Girona 2 s Roman 1st c. AD 240 bp T3 this study ROMN1 Empúrie EM50 Girona 3 s Roman 1st c. AD 316 bp T3 this study ROMN1 Empúrie EM51 Girona 4 s Roman 2nd c. BC 316 bp T3 this study ROMN1 Empúrie EM52 Girona 5 s Roman 1st c. AD 175 bp T/T3 this study ROMN1 Empúrie EM55 Girona 6 s Roman 1st–3rd mosaic c. AD ? this study ROMN1 Empúrie EM57 Girona 7 s Roman 3rd c. AD 316 bp T3 this study ROMN1 Empúrie EM80 Girona 8 s Roman 1st c. BC 316 bp T1 this study ROMN1 Empúrie EM81 Girona 9 s Roman 2nd c. AD 203 bp T1 this study ROMN2 Empúrie EM82 Girona 0 s Roman 3rd c. AD 175 bp T/T3 this study 900– Montsori Barcelon Middle 1200 u a Age AD 240 bp T1 this study MIDD0 IBE2 1 Site MIDD0 MAD4 Cueva de Teruel 2 7 Joaquin Middle 1120 Age AD MIDD0 MAD1 San 3 9 Pablo 1300– Middle 1500 240 bp Age AD Burgos 117 bp + 100 T3 bp T3 Source AY8472 08 AY8472 01 6 MANUSCRIT ACCEPTAT Lab Sampl code e code Site MIDD0 MAD2 San 4 0 Pablo MIDD0 MAD2 San 5 2 Pablo MIDD0 MAD2 San 6 3 Pablo MIDD0 MAD2 San 7 5 Pablo Area Period Date Fragment length Haplogro up Source Burgos 1300– Middle 1500 240 bp Age AD T3 AY8472 02 Burgos 1300– Middle 117 bp + 100 1500 T3 Age bp AD AY8472 03 Burgos 1300– Middle 1500 240 bp Age AD T3 AY8472 04 Burgos 1300– Middle 1500 240 bp Age AD T3 AY8472 05 1. Download high-res image (181KB) 2. Download full-size image 7 MANUSCRIT ACCEPTAT Fig. 1. Location of archaeological Iberian sites by period. (A) Neolithic (5400– 5000 cal. BC): 1, Cova de l'Or, Alicante; 2, Cueva del Mirador, Atapuerca; 3, La Draga, Girona. (B) Bronze Age (1900–1600 cal. BC): 4, Gatas, Almeria; 5, Portalón, Burgos; 6, Valparaiso de Abajo, Cuenca. (C) Iron Age to Roman (5th c. BC to 3rd c. AD): 7, St. Julià Ramis, Girona; 8, Empúries, Girona; 9, Olivet d'en Pujol, Girona; 10, Vila Ametllers, Girona; 11, Vilauba, Girona; 12, Baetulo, Barcelona; 13, Son Fornés, Mallorca. (D) Middle Age (900–1500 AD): 14, Montsoriu, Barcelona; 15, Cueva de Joaquin, Teruel; 16, San Pablo, Burgos. 2.2. Sample extraction and amplification DNA was successfully amplified from 17 of the 20 samples in three laboratories (Cambridge, Dublin and Grenoble) exclusively dedicated to ancient DNA analyses. Extraction, contamination control and amplification reactions were as described previously (Edwards et al., 2004; Campana, 2007), and followed standard ancient DNA practice. For each sample, at least two independent DNA extractions were performed and extraction and PCR negative controls all produced negative results. 252 base pairs (bp) [16,042–16,158 and 16,179–16,313; see Bollongino et al. (2006) for primer details] of the hypervariable control region of the mitochondrial genome were sequenced. Samples from the Roman site of Empúries were amplified with primers detailed in Campana (2007) [namely BT1F–BT3R, BT2F– BT2R, BosCentreF–BosCentreR and BT1F–BTRUTH], which amplified a longer 316 bp sequence. Five of the 11 samples from Empúries that had endogenous DNA generated this full 316 bp fragment (Table 1). Inter-lab replication, including sample preparation, extraction and PCR, was undertaken on four samples (NEOL03, BRON01, ROMN06 and MIDD01) and the sequences obtained were consistent across laboratories. 2.3. Data analyses Sequences from 25 published archaeological cattle were included in the analyses (Anderung et al., 2005; Colominas et al., 2014) to give a total dataset of 42 ancient cattle sequences from Iberia. This is the largest dataset presented from this region. Moreover, a total of 1130 mtDNA sequences from European and African Bos taurus (from GenBank) were used to compare archaeological cattle diversity to that of the present day. Details of the number of individuals, breed of origin and geographical areas are shown in Table S1. The mtDNA sequences were aligned in MEGA (ver. 6; Tamura et al., 2013) to the reference sequence published in Troy et al. (2001), and all were truncated to 240 bp. Median-joining networks were constructed following Bandelt et al. (1995). A three-dimensional statistical parsimony network was constructed using the R script TempNet (found at http://web.stanford.edu/group/hadlylab/tempnet/; Prost and Anderson, 2011). In addition, analyses of inter-population genetic distances between extant and ancient populations were performed in ARLEQUIN (ver. 3.5; Excoffier et al., 2007) using published modern regional data from native breeds (Table S1; Cymbron et al., 1999; Troy et al., 2001; Miretti et al., 2002; BejaPereira et al., 2006; Pellecchia et al., 2007; Ginja et al., 2010) in ARLEQUIN (ver. 3.5; Excoffier et al., 2007). 8 MANUSCRIT ACCEPTAT 3. Results Amplification of a 240 bp section of the control region was successful in 13 of our samples, with a further four samples yielding sequence data of less than 240 bp in length. The sequences were aligned against a highly polymorphic region of the control region as characterised by Troy et al. (2001) (Table 2). All archaeological samples gave sequences similar to those encountered in modern native cattle. The dominant feature among the archaeological samples was the predominance of the T3 haplotype (eight samples). Five individuals had identical sequences to the T3 reference sequence (NEOL01, NEOL02, ROMN11, ROMN13 and ROMN17). In addition, a further three individuals grouped within T3 haplotypes, but with one or more nucleotide substitutions (ROMN09, ROMN12 and ROMN14), and one sample (BRON01) had a T haplotype. Three samples, where the full sequence could not be generated (ROMN08, ROMN15 and ROMN20), could only be classified as T/T3, due to the non-amplification of the diagnostic 16,255 position. Table 2. Variation in mitochondrial control region sequences. The variable positions in control region sequences of archaeological cattle samples aligned to the European consensus haplotype (T3). Differences are indicated, with a period (.) denoting identity. Sequence codes from Table 1 are given in the first column and only variable sites are shown. The sequence positions from the BOVMT GenBank sequence are given above each column (accession number V00654; Anderson et al., 1982). The common and putatively ancestral Middle Eastern and European consensus sequences are denoted T and T3, with T1 and T2 denoting the consensus sequences from Africa and the Middle East/Europe, respectively. All ancient sequences generated as part of this study are shown in bold type, and each sample has been assigned to one of the four main haplogroups by means of its relative position in the median-joining network (Fig. 2). Five individuals belonged to haplogroup T1 (Table 2). One individual had an identical sequence to the T1 reference sequence (ROMN18) and four others exhibited a T1 haplotype (NEOL03, ROMN06, ROMN19 and MIDD01). One individual is from the Neolithic period (La Draga), three are from Roman 9 MANUSCRIT ACCEPTAT chronologies (Son Fornés and Empúries) and one is from the medieval period (Montsoriu) (Table 1; Fig. 2). 1. Download high-res image (536KB) 2. Download full-size image Fig. 2. Median-joining network comparing 13 novel and 16 published archaeological cattle sequences with extant native North African and Iberian breeds. Median-joining network drawn using 240 bp fragment to compare the 29 archaeological samples (13 novel and 16 previously published; Table 1) with 159 individuals from 9 modern North African native breeds and 447 individuals from 24 modern Iberian native breeds (Table S1). Circles are coloured according to which haplogroup the sequences belong to: blue = T; yellow = T1; green = T2; red = T3. Ancient samples are highlighted – T and T3 ancient haplotypes are shown in white, and T1 ancient haplotypes are shown in black. Notable haplotype assignments are indicated (labelled as in Table 2), with N = Neolithic, B = Bronze Age, R = Roman, and M = Middle Age. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) These data were compared with published archaeological mtDNA sequences from Iberia (Table 1), establishing a data set of 42 ancient cattle sequences from different time periods, the largest dataset presented from Iberia. From this published data, one individual had a T1 haplotype, a sample from the Bronze Age site of Portalón (Anderung et al., 2005). All other individuals, dating from the Neolithic to the medieval period, had a T/T3 haplogroup. The T2 haplogroup was 10 MANUSCRIT ACCEPTAT not represented in the data set. The haplotypic diversity values of cattle populations from Roman Empúries and Middle Ages San Pablo sites (Table 3; Fig. 3) revealed large diversity in these two time periods. In all time periods tested (Neolithic, Bronze Age, Roman and Middle Ages), haplotypes were found to belong to both the T1 and T3 haplogroups (Table 1; Fig. 2). 1. Download high-res image (229KB) 2. Download full-size image Fig. 3. TempNet showing all ancient Iberian samples. Temporal median-joining network drawn using 240 bp fragment from all ancient Iberian cattle. Circles represent sequence haplotypes, the area being proportional to the haplotype frequency. Points are theoretical intermediate nodes introduced by the medianjoining algorithm, and branches between haplotypes represent single nucleotide mutations, unless otherwise indicated. Circles are coloured by the age of the samples: red = Neolithic; bright green = Bronze Age; turquoise = Roman; purple = medieval. Empty nodes at each time frame indicate the absence of that haplotype at that period of time. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Table 3. Genetic diversity values of archaeological cattle. Genetic diversity of the archaeological Spanish cattle populations from Bronze Age Portalon, Roman Empúries and Middle Ages San Pablo, using the 240 bp fragment of the control region. Country Site Period Number of % Samples per haplogroup Number of haplotypes Haplotype diversity 11 MANUSCRIT ACCEPTAT samples SPAIN Portalón Bronze 11 Age 3 Empúries Roman 7 GROUPS San Pablo Middle 4 Ages T T1 T2 T3 9.1 9.1 14.3 81.8 3 0.346 ± 0.172 85.7 5 0.857 ± 0.137 100 3 0.833 ± 0.222 The archaeological samples from the Iberian Peninsula were compared with mtDNA sequences from European and African Bos taurus sequences from GenBank (Table S1) to compare past cattle diversity. As can be seen from the median-joining network (Fig. 2), the archaeological data fit within living cattle diversity. The T1 sequences are interesting because the closest matches for these individuals in the modern data set are from a broad geographical range. A complex history of population movements may be reflected in these samples with a number of potential origins for these individuals. NEOL03 matched an extant individual from Guinea, ROMN06 and ROMN19 matched extant individuals from Morocco with 99% homology, and MIDD01 matched an extant individual from Tunisia (Table S2). 4. Discussion Our analyses show that haplogroup T3, which dominates among modern domestic European cattle, was the most frequently occurring haplogroup in Iberia since the Neolithic. However, our data also show a presence of T1 haplotypes in Iberia from the Neolithic period through the Bronze Age into the Roman period and the Middle Ages. The presence of T1 haplotypes in Neolithic Iberia, and their chronological and spatial recurrence, clearly demonstrates that T1 was not solely a Muslim or later introduction, as some have argued (Cymbron et al., 1999, 2005; Beja-Pereira et al., 2002, 2003, 2006; Pérez-Pardal et al., 2010), though additional and/or new T1 haplotypes may have been introduced during these periods. The Neolithic T1 cattle in our data most likely represent populations that expanded out of the Near East in the company of early farmers moving along the Mediterranean coastal route. T1 haplotypes could have reached the coast of eastern Iberia from the Near East as early as ∼5400 cal. BC (Price, 2000; Zilhão, 2001), when human migrations and contacts through the Mediterranean basin are archaeologically attested (Winiger, 1998; Gkiasta et al., 2003; Berrocal, 2012). Domestic cattle are well documented along the western Mediterranean (Tresset and Vigne, 2007) and coast of Spain (Saña, 2013) at this time, with La Draga being one of the earliest Neolithic Iberian sites where domestic cattle are found (Saña, 2000). The site of La Draga has close connections with the Mediterranean route of diffusion of the Neolithic, both culturally (Saña, 2013) and due to its location on the eastern coast of the Peninsula (Fig. 1). As the earliest cattle in the coastal areas of north-west Africa are dated significantly later than the earliest Neolithic farming 12 MANUSCRIT ACCEPTAT sites found there (Jousse, 2004; Mulazzani, 2013; Manen, 2014), it is unlikely that cattle could have arrived at La Draga via an African route. The presence of the T1 haplotype at La Draga implies, therefore, that this haplotype travelled with the earliest wave of domestic cattle from the Near East to the north-western Mediterranean. The contacts between the Iberian Peninsula and the rest of the Mediterranean continued during the Bronze and Iron Ages (Ruiz-Gálvez, 1993; Stampolidis, 2003; Celestino et al., 2008), and during Roman and medieval times, with the development of a large and regular sea trade along all the Mediterranean Sea (Rauh, 2003; Arnaud, 2005; Tchernia, 2012; Broodbank, 2013). The large diversity seen in the Roman mtDNA sequences are suggestive of more intensive contacts during this period (Fig. S1). Empúries was a trading post since the 6th century BC (Aquilué et al., 1999), and the genetic distinctness of the haplotypes found there may either represent a local maintenance of cattle introduced during the Neolithic from the Near East, or new introductions, from other regions of Europe or directly from Africa. Although T1 is seen at low frequencies in extant Iberian and Italian breeds, our results suggest that the haplogroup was lost from the majority of European populations sometime after the Middle Ages. 5. Conclusions Domestic animals have been one of the key pillars of social and economic change during the last ten millennia. Our results show the presence of the two most common haplogroups in domestic cattle in Europe (T3) and Africa (T1) in three early Neolithic sites in Iberia. These haplogroups are also documented in Iberia during the Bronze and Iron Ages, as well as in the Roman and Medieval periods. The absence of the T2 haplogroup, which has an appreciable frequency in several Eastern European and Italian cattle breeds (Bradley et al., 1996; Beja-Pereira et al., 2006; Pellecchia et al., 2007; Achilli et al., 2009), is striking and requires special attention in future studies. Our results provide important data about the phylogeny of cattle haplogroup T1 in the Iberian Peninsula, and open new issues about cattle movements through time. Further studies of the geographical distribution of the T1 haplogroup in Europe prior to ∼5400 cal. BC, when the Neolithic arrived into the Iberian Peninsula, would shed more light on this topic. In addition, we would suggest further studies concentrate on analysis of Y-chromosomal diversity, thus obtaining paternal information about cattle movements in the past, and a systematic DNA analysis of Mesolithic aurochs in Europe, especially in the south of the continent. Acknowledgements For providing access to the archaeological samples, we thank J. Tarrus, A. Bosch and J. Chinchilla [La Draga; work funded by Ministerio de Economía y Competitividad, Spain: HAR2011-25826]; V. Lull, R. Micó, C. Rihuete and R. Risch [Gatas and Son Fornés; given in the frame of the project ‘Circulación y movilidad en el sureste ibérico durante El Argar (ca. 2200–1550 cal. BC): relaciones de distribución y delimitación de territorios políticos y económicos’: HAR2011-25280]; 13 MANUSCRIT ACCEPTAT and J. Mateu, J. Tura, G. Font, S. Pujadas, J. M. Llorens and Museu Etnològic del Montseny [Montsoriu]. L. Colominas was funded by a postdoctoral fellowship (HA2010-0293) from the Government of Spain (‘Programa Nacional de Movilidad de Recursos Humanos del Plan Nacional de I + D + i 2008–2011’) and by the McDonald Grants & Awards Fund 2013, McDonald Institute for Archaeological Research, University of Cambridge. C. J. Edwards is currently supported by a Leverhulme Trust research project grant (project no. RPG-388), but would like to recognise previous support from the Irish Research Council for Science Engineering and Technology (project numbers SC/1999/409 and SC/2002/510), The Wellcome Trust (grant no. 047485/Z/96/Z) and from a Eurocores OMLL Programme Grant via CNRS, France. M. A. Bower was supported by the McDonald Institute for Archaeological Research and the University of Cambridge. Appendix A. Supplementary data The following is the supplementary data related to this article: Download Acrobat PDF file (99KB)Help with pdf files References      Achilli et al., 2008 A. Achilli, A. Olivieri, M. Pellecchia, C. Uboldi, L. Colli, N. Al-Zahery, M. Accetturo, M. Pala, B. Hooshiar Kashani, U.A. Perego, V. Battaglia, S. Fornarino, J. Kalamati, M. Houshmand, R. Negrini, O. Semino, M. Richards, V. Macaulay, L. Ferretti, H.-J. Bandelt, P. Ajmone-Marsan, A. TorroniMitochondrial genomes of extinct aurochs survive in domestic cattle Curr. 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