The universal history of Jewish people is full of constant oppressions and persecutions Portugal as no exception to this rule It was in TrasosMontes a rugged and isolated province of Portugal on the North East border with Spain that many Jews settled and were able to secretly maintain their ancestral culture and religious practises for more than four centuries This Marrano culture of inestimable value has been the object of a few scattered studies by ethnologists and historians However there was yet to occur any modern genetic research that could shed the light of science on the historical knowledge that has already been gathered To fill this gap was our main aim of research carried out by IPATIMUP Institute of Molecular Pathology and Immunology of the University of Porto and the University of Coimbra

A little history

The first archeological evidence of the presence of Jews in Portugal dates back to the fifth century in a tombstone unearthed in Mertola where a menorah is shown (Pignatelli 2000; artins 2006) From the beginning of the monarchy up to the XV century there were tolerant kingdoms in which the Jewish communities proliferated as well as others which were less forbearing where numerous restrictive measures were taken such as the prohibition of access to public positions the use of distinguishing items of clothing or obligatory curfews (Paulo 1985; Canelo 1987; Pignatelli 2000; Martins 2006)

Jews probably established themselves in TrasosMontes after 1187 because of the benefits offered to them by the foral of king D Sancho I During the more tolerant reigns Jewish communities spread throughout Portugal A highlight occurred during the reign of king D Joao II who put at his service Jewish doctors mathematicians and cosmographers who played a fundamental role in the Portuguese Discoveries (Pignatelli 2000; Martins 2006)

In 1492 the Decree of Expulsion was issued in Spain It prompted a massive emigration to Portugal estimated to be above a hundred thousand people (Carvalho 1999) It is thought that at least three thousand Spanish Jews crossed the border into TrasosMontes (Alves 1974) There followed a rapid expansion of local commercial activities such as tanning shoe making iron mongering and later the silk industry At this time there is evidence of the existence of a synagogue within the walls of the castle of Braganca capital of the district This indicates the importance that the local Jewish community had during this period (Jacob 1997)

In December 1496 the Portuguese King also signed a decree of expulsion However this resolution was modified in May 1497 by means of a forced conversion in which about twenty thousand Jews from all of the country who were preparing themselves to depart in exile were baptized against their will This attempt at a political solution through integration did not have the desired effect Tensions and hostilities between conversos and oldChristians became more and more accentuated and the phenomenon of cryptoJudaism emerged (Saraiva 1985; Canelo 1987; Mea 1997; Pignatelli 2000; Martins 2006) In May 1536 a papal bull was issued which established the Inquisition in Portugal In 158283 the Inquisition arrived in TrasosMontes reaching a peak of activity in 1599 Accusations and imprisonments followed and the list of names of individuals charged with Judaism grew day by day (Azevedo 1994; Mea 1996; Mea 1997) An example of these grim times is found in the village of Carcao where among 150 families 130 people were arrested for Judaism many of whom perished in the fires of the Inquisition (Andrade & Guimaraes 2008)

In the XVII and XVIII centuries the inquisitorial process was intensified and as a consequence there was a significant exodus to other countries particularly from the manufacturing and mercantile elite which then were flourishing in this region (Paulo 1985; Garcia 1993) At the end of the fifteenth century there were approximately 134 Jewish communities throughout the country with a population estimated at 100 thousand which reflect 10 percent of the total population The exact number of those who emigrated is not known however it is believed that in 1631 the Jewish population was reduced to 10 thousand (Carvalho 1999) Initially in the midsixteenth century many of them set up in Amsterdam London Hamburg and some French and Italian cities After that some of them went to the new Portuguese colonies in Africa India and Brazil and then from the cities of north Europe to the New World Curacao Paramaribo and the US (Carvalho 1999; Pignatelli 2000; Martins 2006) The major Diaspora of crypto Jews actually begins in the fifteenth century and continued up until the eighteenth century at which point the Pombaline discrimination law came into effect and the official persecution of the Inquisition ended (Carvalho 1999; Martins 2006) It is not until the XIX century that the Jewish communities’ resurgence was attested In 1927 the Jewish community was founded in Braganca and in June 1928 the Synagogue “Shaaré Pideon” was inaugurated 431 years after the previous one During the Dictatorship period from the year 1934 on the community of Braganca gradually disaggregated The descendants of this extinct community have remained dispersed through the district area still claiming Jewish identity although most of them have been assimilated into the local culture and even in some cases into the dominant religion

A little genetics

Patterns of genetic diversity depend on one hand on history which includes demographic phenomena of migration substructure and fluctuations in the population size and on the other hand of genetic factors such as mutation rates and selection (Agrafioli & Stumpf 2007) The understanding of this complex evolutionary process has in recent years deserved particular interest given its practical application in fields such as evolutionary genetics medicine etiology of human diseases forensic studies and genealogical reconstructions (Hammer et al 2001; Brion et al 2005; Agrafioli & Stumpf 2007) The Y chromosome is passed exclusively from father to son without recombination (with the small exception of its tips) thereby providing a model of excellence for the study and understanding of genetic diversity (Jobling & Tyler Smith 2003)

The information contained in genetic diversity is obtained by using different molecular markers or polymorphisms in DNA sequences Markers as STRs (for Short Tandem Repeats) mutate more quickly causing a greater degree of variability and therefore are used in the reconstruction of recent events ( Butler 2003; Payseur & Cutter 2006; Agrafioli & Stumpf 2007) Other polymorphisms as SNPs (for Single Nucleotide Polymorphisms) due to their low rate of mutation represent the most appropriate markers for defining stable paternal lineages over time periods of thousands of years (Agrafioti & Stumpf 2007) Haplogroups are defined as a linear combination of states at SNPs Within each of these haplogroups are sub strains determined by variations of STRs which are called haplotypes (Knijff 2000; Y Chromosome Consortium 2002)

Genetic studies concerning the origin of the Jewish people are not new There are an extensive number of articles at the NCBI database (National Centre for Biotechnology Information) Initially they started by using classical genetic markers such as blood groups More recently there have been a number of studies with markers of the Y chromosome (male lineages) and mtDNA (female lineages) which have not only helped to establish genetic relationships between different communities but have also unveiled details of their origin and migr ation patterns

Our research

A combination of geographic religious ethnohistorical and individual affiliation criteria were used to select the population samples Buccal cells were collected on cytology brushes from sixty unrelated selfdesignated Jewish males from several villages such as Argoselo Carcao Vilarinho dos Galegos Mogadouro and Braganca in TrásosMontes and also from Belmonte All samples were taken under informed consent Personal inquiries were made to each individual in order to confirm their Jewish ancestry and to avoid close kinship (As strange as it may seem in some of the more hidden away villages of TrásosMontes there was found even today some reluctance to admit to Jewish ancestry which only reflects the power of four centuries of antiSemitism and official persecutions)

In order to characterize the genetic composition of these Portuguese Jews we preformed a highresolution Ychromosome STR and SNP typing strategy Twentythree YSNPs were typed in order to define the major haplogroup lineages while sixteen STRs were analysed in order to characterise haplotypes

A high genetic diversity was found at SNP level with eleven different identified haplogroups Three of them occur with higher frequencies making up a total of 83 percent of the sample collected Another six haplogroups appeared with much lower frequencies representing in some cases only one individual therefore most of them do not merit further discussion in this article

The haplogroup found at the highest frequency is also observed in the nonJewish Portuguese population but at much lower proportions 29 percent vs 58 percent (Beleza et al 2006) and it has not been referred to in studies of other Jewish populations until recently (Adams et al 2008) This haplogroup emerges as the most frequent haplogroup in the Iberian Peninsula and the Western European coastline displaying a increasing distribution gradient moving from east to west (Semino et al 2000; Bosch et al 2001; Cruciani et al 2002; Flores et al 2004; Brion et al 2005; Moore et al 2006) These facts could indicate either some degree of miscegenation with the Portuguese/Iberian host population or miscegenation during the Diaspora with other Western European populations In contrast one of the most frequent lineages found in Ashkenazim Jews displays low frequency in our sample in accordance with the fact that Ashkenazim Jews mainly came to Portugal during and after the Second World War and even then they settled almost exclusively in Lisbon and the Algarve

Another very frequent (39 percent) haplogroup has its origin in the Middle East and its pattern of distribution follows a decreasing gradient from there to Western Europe This fact can be linked either to the diffusion of the Neolithic farmers (Underhill et al 2001; Semino et al 2004) or later maritime migrations throughout the Mediterranean basin such as the Phoenician (Hammer et al 2000; Di Giacomo et al 2003) It would appear therefore that the elevated frequencies encountered here can be explained by the historical migration of the ancestral Sephardic population to the Iberian Peninsula which according to some historians accompanied the Phoenician trade routes

The last frequent haplogroup in the Jewish population studied here reached the remarkable frequency of 15 percent contrasting to that found in the general Portuguese population 16 percent (Beleza et al 2006) It should be noted that this is a rather rare haplogroup with a global frequency of around 1 percent (King et al 2006) however it is has quite high frequencies in general Sephardic Jews 23 percent and Israelis 13 percent (Behar et al 2004; Shen et al 2004) These frequencies since they appear with similar values in Israeli Sephardic Jews could demonstrate a clear link to the original Sephardic population and probably would have constituted part of the original genetic patrimony of this group Supporting this idea is the relatively low frequencies with which this haplogroup emerges along the Mediterranean coast

The haplogroup frequencies determined in this work were used to analyse genetic distances between this group and other Jewish populations as well as their respective host populations (Behar et al 2004; Shen et al 2004; Beleza et al 2006; Adams et al 2008) What emerged was the fact that Jews of TrasosMontes share more genetic affinities with Jewish populations of Europe and the Middle East particularly other Sephardic Jews than with the nonJewish Portuguese population in general This is consistent with other studies into diverse Jewish populations and their host communities around the world

The analysis of the fast evolving markers (STRs) shows a little higher haplotype diversity than that found for the general Portuguese population The conclusions that can be drawn are in the genetic YSTR pool that we studied there are neither strong drift nor founder effects acting This population probably resulted from a combination of a number of different lineages which could reflect the historically constant flux of Jewish populations from other areas over the last five hundred years associated with a stable size within this community

The high diversity found both at haplotype and haplogroup levels (9877 percent and 0822+/0025 respectively) is rather surprising in a demographically small and inbred community A deeper and more detailed investigation is required in order to clarify how these communities avoided the genetic erosion caused by over four centuries of religious repression

For the most frequent Near East/Jewish haplogroups found here we used STR information to conduct a haplotype match analysis using YHRD the Y Chromosome Haplotype Reference Database (wwwyhrdorg) Curiously coincidences were found in such diverse locations as Northern Europe USA and Latin America This probably reflects what is known in terms of the history of the Diaspora from the Iberian Peninsula

All this new data makes up no more than the first steps on a long road of research which will eventually bring to light in much greater detail the complex history of the Iberian Jewish Diaspora In the full awareness of this our research team is now setting out to analyze the second Diaspora of Jewish populations of Iberian origin in Northern Europe USA and Latin America using the Y chromosome genetic polymorphisms briefly reported here along with those that define the maternal lineages the mtDNA In addition recombining autosomal and X chromosome markers will be typed as well to trace the demographic history of these communities

This endeavour will require the active involvement of such local communities and cooperation in sample collection is essential To that end specific diffusion actions targeted at the explanation of project aims and background will soon be undertaken allowing for an effective informed consent on an individual and collective basis Furthermore it should be noted for those interested that at this moment the team is finalizing an article which will discuss the technical details of the research done so far

Ináªs Nogueiro Departamento de Antropologia Universidade de Coimbra 3000 Coimbra Portugal Leonor Gusmëo IPATIMUP Instituto de Patologia e Imunologia da Universidade do Porto R Dr Roberto Frias s/n 4200465 Porto Portugal António Amorim Faculdade de Ciáªncias da Universidade do Porto 4050 Porto Portugal


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