research article paleogenetic studies in guajajara

Research ArticlePaleogenetic Studies in Guajajara Skeletal RemainsMaranhatildeo State Brazil

Daniela Leite1 Alysson Leitatildeo1 Ana Paula Schaan1

Anderson N R Marinho1 Sheila Souza2 Claudia Rodrigues-Carvalho3

Francisca Cardoso45 and Acircndrea Ribeiro-dos-Santos1

1 Laboratorio de PaleogeneticaLaboratorio de Genetica Humana e Medica ICB Cidade Universitaria Prof Jose da Silveira NettoUFPA Rua Augusto Correa 01 Guama 66075-970 Belem PA Brazil

2 ENSPFiocruz Avenida Brasil 4365 Manguinhos 21040-900 Rio de Janeiro RJ Brazil3Museu NacionalUFRJ Quinta da Boa Vista Sao Cristovao 20940-040 Rio de Janeiro RJ Brazil4 Instituto de Filosofia e Ciencias Humanas (IFCH) UFPA Rua Augusto Correa 01 Guama 66075-970 Belem PA Brazil5 CRIA FCSH Universidade Nova de Lisboa Campus de Campolide 1099-085 Lisboa Portugal

Correspondence should be addressed to Andrea Ribeiro-dos-Santos akelyufpagmailcom

Received 12 November 2013 Revised 3 April 2014 Accepted 9 April 2014 Published 14 May 2014

Academic Editor Santos Alonso

Copyright copy 2014 Daniela Leite et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

In the early 17th century French and Portuguese colonizers and Jesuit priests settled in the state of Maranhao and made contactwith the Guajajara an ethnic group that lived along the margins of the Pindare River The Guajajara maintained contact withBrazilian national society over the centuries including with Brazilian admixed populations and with African slaves that flockedtowards the region from the 18th century onwardsThe present study investigates the origins of this admixture usingmitochondrialgenetic variabilityThe bones of 12 individuals investigated which are currently part of the collection of theNational Museum weretested for genetic diversity aDNAwas extracted by the phenol-chloroformmethod and by DNA IQ (PromegaMadisonWI USA)Amplification of the HVS I region was performed by PCR followed by direct sequencing using the Big Dye kit (Life TechnologiesFoster City CA USA) This region was found to represent haplogroups of Amerindians (A C and D) and Africans (L L1b L1cand L3) The presence of African haplogroups in Guajajara bones from as early as the 18th century is consistent with historicaland anthropological data suggesting the admixture with Africans andor AfrodescendantsTherefore this study demonstrates thatwomen with African haplogroups were introduced into the Guajajara population

1 Introduction

11 The Guajajara Among the indigenous peoples that stillinhabit the vast South American continent are the Guajajaraan example of ethnic resistance after four centuries of contactwith European Brazilian and African populations TheGuajajara are also referred to as the Tenetehara-Guajajara ofMaranhao to distinguish them from the Tenetehara-Tembe ofPara and they belong to the Tupi linguistic family Accordingto Nimuendaju [1] during the 19th century the Guajajaralived along the Pindare Grajau andMearim rivers (Figure 1)an area that they had occupied since pre-Columbian timesaccording to Wagley and Galvao [2]

A more detailed analysis of the populationrsquos demographyrevealed that although their population size had been severelyreduced in the past it recovered significantly during atleast two historical periods and is an extraordinary exampleof demographic expansion Another particularity of thesegroups is their substantial level of admixture [3] both withAfricans and Europeans Additionally in the contemporarydemographic process it is noticeable that more Guajajaramen have married women of the surrounding nationalsociety than women have married non-Guajajara men Thegreater mobility of men in search of work and the socialpressure for the permanence of women within the matrilocalvillages help to explain this pattern According toWagley and

Hindawi Publishing CorporationJournal of AnthropologyVolume 2014 Article ID 729120 8 pageshttpdxdoiorg1011552014729120

2 Journal of Anthropology

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1∘09984000998400998400S

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3∘09984000998400998400S

4∘09984000998400998400S

5∘09984000998400998400S

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10∘09984000998400998400S

Imperatriz

Guajajara

Quilombo

tribes

populationsSatildeo Luiacutes

Figure 1 Current distribution of the studied Guajajara villages ofKamiranga and Januaria (in red) and the quilombola populations(in black) in the state of Maranhao Brazil (adapted from [59])

Galvao [2] and Gomes [3] the Guajajararsquos traditional kinshipsystem is flexible enough to accommodate the need for seek-ing marriages in more distant and less related communitiesthereby acting as a buffer during the most critical demo-graphic periods The historical conditions of admixture withother ethnic groups throughout four centuries of contacts aremapped in the historical-anthropological revision by Gomes[3] However the precise time that the first events of contactwith African andor Afrodescendants started is not clear inthe historical data (Box 1)

12 Bioanthropology and Bioarchaeology of Guajajara TheGuajajara are currently one of the most documented ethnicgroups of Brazil Many studies have recorded the ethnog-raphy demography history bioanthropology and bioar-chaeology of the Guajajara [4ndash7] Lima [4] documentedthe interesting practice of intentional dental modificationprevalent among the Guajajara and it was confirmed in someexhumed skulls The dental modification was characterizedby symmetric cuts of the lateral and medial angles of theincisors called ldquopiranha teethrdquo and was attributed to theadoption of African habits This interpretation was based onoral information about Afrodescendants from some of theGuajajara in the villages Other biological features other thansome adopted material culture music and customs of theseIndians were also recognized as being of Afro-Americanorigin In the past dental modification was found in severalregions of Africa In Brazil other indigenous groups such

as the Tucuna Makuxi Galibi Karipuna Pauaxiana Jauariand Kaigang also adopted similar dental modifications forthe same reasons since the practice of intentional dentalmodification has never been demonstrated in Brazilianarcheological sites prior to contact [4 8ndash10]This practice hasalready been described amongAfricans andAfrodescendantsin Brazil [9 11 12]

The distribution of genetic patterns of the ancestralmtDNA complements and confirms the results obtained bymorphological and archeological studies it has enabled theformulation of a more adequate model to explain prehistoricdiversity and the peopling of the Americas These char-acteristics confer significant advantages of the use of themitochondrial genome for the research in the present studyHowever one must not forget a significant limitation froma biological perspective it only narrates one part of humanhistory that of the maternal lineage

The specificity of mtDNA lineages among different popu-lations and groups has favored their use as ancestry markersover the past few decades [13ndash20] Among Amerindianpopulations five main haplogroups are prevalent (A B CD and Xmdash[21 22]) [13 23ndash27] Asian populations shareAmerican haplogroups (A B C and D) as well as otherhaplogroups (M N G F Z Y and R) that are described asspecific to the Asian continent [28 29] European and Euro-Asian populations also possess more frequent haplogroups(I J K H T U V and W) [15 28 29] In contrast Africanpopulations have a large amount of exclusive haplogroupsand subhaplogroups (L0 L1 L2 L3 L4 L5 and L6) [18 3031]

This study is a first step toward elucidating the geneticcharacteristics of the Guajajara individuals Our objective isto characterize the admixture with non-Amerindian popula-tions In addition to contributing to our understanding of thehistory of this ethnic group this study aims to corroborateprevious bioanthropological studies and to contribute tothe knowledge of regional population dynamics during thecenturies that followed colonization until the final formationof what is today the Brazilian national society The presentstudy uses skeletal samples from the Guajajara cemeteriesto investigate aDNA mitochondrial haplogroups and tocontribute to the reconstruction of part of its history

2 Materials and Methods

21 Subjects The bone samples belong to the BiologicalAnthropology collection of the National Museum Rio deJaneiro The samples were obtained from 12 different indi-viduals exhumed from cemeteries of the Guajajara villages ofJanuaria and Kamiranga (Figure 1) These skeletons are partof the collection brought by Pedro Lima and his collaboratorsin 1945

The bone samples were collected from ribs removed fromeach individual The ribs were selected following criteria ofintegrity absence of pathological signs anomalies or severetaphonomic changes that could compromise future inves-tigations about the health state of other parts of the skeletons(bioarchaeological studies such as age and sex assessment or

Journal of Anthropology 3

First colonization try at Maranhao 1553French invade Sao Luiz island they contact ldquoPinariensrdquo (Maranhao) 1612Portuguese expel the French 1613Portuguese refer the Guajajara Amerindians (Maranhao) 1615Jesuıts fund the first cathechist missions (Maranhao) 1653Ships bringing the first Africans slaves Sao Marcos bay (Maranhao) century VIIFoundation Mission Carara (Maranhao) century XVIIIFirst slave ships officers arrive (Maranhao) 1761Refuge of Cabanagem soldiers (Grao-Para) 1835ndash1841Foundation of the original village Januaria (Maranhao) 1854Migration of refugees from drought in the Northeast 1877ndash1880Foundation of the Quilombos in Imperatriz (Marnhao) century XIXFoundation of the present village Januaria century XX

Box 1 Chronology related to contact with the Guajajara (upper Pindare River)

Table 1 Sex age and village of origin of the studied samples and a description of analyses performed in each methodological stage

Sample Sex Age Number of extractions Number of sequences Sequences obtained VillageGAJ 700 Female 1820 2 21 11 KamirangaGAJ 701 Female 2025 2 17 4 KamirangaGAJ 709 Female +50 2 21 6 JanuariaGAJ 713 Undetermined 612 2 13 6 JanuariaGAJ 715 Female +20 2 21 6 JanuariaGAJ 7171 Male 2049 2 10 11 JanuariaGAJ 7182 Male 2034 2 15 7 Januaria1Skeleton number GAJ 717-MN-RJ provided a 19th century chronology of 140 plusmn 30 BP (BETA-291714-AMS C13 corrected) 2Skeleton number GAJ 718-MN-RJ provided an 18th century chronology of 210 plusmn 40 BP (GEOCHRONMA GX31824-AMS C13 corrected)

other additional information) The upper ribs in particularthe fourth rib were preserved as they are used for ageassessment and therefore contain additional bioarchaeologi-cal data All of the selected bone samples were photographedand described prior to being sent to the laboratory foranalysis

Osteometric and osteological data as well as sex andage were already estimated in previous studies and theinformation in the National Museum archives was used forthe present study (Table 1) The material was also dated forthe purpose of the present investigation the skeleton number717-MN-RJ provided a 19th century chronology of 140 plusmn30BP (BETA-291714-AMS C

13corrected) and the skeleton

number 718-MN-RJ provided an 18th century chronologyof 210 plusmn 40BP (GEOCHRON MA GX31824-AMS C

13

corrected)

22 Extraction PCR and Sequencing The extraction andtreatment of the DNA samples were performed in thePaleogenetic section of the Human and Medical Genet-ics Laboratory at the Federal University of Para (UFPA)according to the established protocol [25 32] The DNA wasextracted either with the phenol-chloroform method [33]withmodifications or with the DNA IQ System kit (PromegaMadison WI USA) The method used depended on thestate of preservation of the samples The extracted materialwas then stored in a freezer until PCR amplification of thehypervariable region I (HVS-I ranging from 15920 to 16498)

For this purpose two sets of primers pairs with overlappingregions were selected which together amplified a 578-bpregion [25 32] A negative control containing all necessarycomponents for a reaction except DNA was used in all thestages of the amplification

The PCR amplification product was sequenced followingthe method of Sanger et al [34] with the Big Dye Termi-nator Cycle Sequence kit which uses AmpliTaq DNA Poly-merase (Life Technologies Foster City CA USA) The directsequencing of mtDNA was performed in an ABI Prism-3130genetic analyzer (Life Technologies) These sequences wereexported and printed using the Chromas v14 software

All steps from extraction to sequencing were repeatedbetween two and five times To confirm the results and toassess the reliability of the sequencing each sample was onaverage extracted 18 times enlarged by PCR 46 times andsequenced 157 times using the two sets of primers in bothdirections (forward and reverse)

The result of each sample was compared to the mtDNAreference sequence [35] to identify the haplogroups Com-parisons were also made with the sequences of each researchparticipant to discard the possibility of contamination frominside the laboratory

23 Data Analysis Neighbor-joining (NJ) trees of mtDNAsequenceswere constructedwithMEGAversion 50 using theKimura two-parameter distance method and the NJ methodof Saitou and Nei [36] bootstrapped with 10000 replicates


In addition the samples were subjected to the analysis ofinconsistent mutations (phantoms) using the Netmat andNetwork v 4109 programs [37ndash39] which were also used toassess the quality and reliability of the obtained results

3 Results and Discussion

The total number of analyzed samples generated 73 sequen-ces 23 of which were excluded either because of contami-nation or because they did not generate a conclusive resultconcerning the HVS-I region of the mtDNA Among the 12individuals analyzed in the present study five were excludedby contamination or by inconclusive results (GAJ 699 GAJ702 GAJ 704 GAJ 705 GAL 711) (Table 1)

To ensure the reliability of the results we followed theauthentication criteria suggested by Cooper and Poinar [40]Gilbert et al [41] Hofreiter et al [42] and Paabo et al[43] for results obtained in ancient DNA sequencing Theyare (1) isolation of the work area the DNA extraction mustbe in a separate area from the PCR amplification (2) theremust be negative control samples to detect the presence ofcontaminants (3) respect the molecular state of the sampleamplify small fragments due to the difficulty of amplifyinglarge fragments (4) reproducibility extractions and PCRsmust be repeated (5) cloning to evaluate the presenceof contaminants (6) independent replication of the resultsin another laboratory (7) morphological and biochemicalpreservation of the material bone structure and biomoleculepreservation aspects that relate to the integrity of the DNA(collagen amino acids etc) (8) quantification to assess thequantity of the extracted DNA Out of these criteria theonly one we did not follow was item number 6 (independentreplication of the results in another laboratory)

In the seven samples that allowed sequencing analysishaplogroups characteristic of Amerindian and African pop-ulations was observed Of the Amerindian individuals onebelonged to haplogroup A another to haplogroup C1 andone to haplogroup D Of the individuals of African originone belonged to haplogroup L two to haplogroup L1 (L1b eL1c) and another to haplogroup L3e (Table 2) None of theindividuals studied belonged to Amerindian haplogroup B orto the European haplogroups

A reticulation was obtained without filter application andcan be explained either by the existence of two differentancestries (Amerindian and African) or by the presence ofhotspot mutations This result was confirmed when the samesamples were analyzed after application of a hotspotmutationfilter (in the Netmat program) as suggested by Bandelt et al[44] followed by the analysis of the Network program

In the present study the samples were aligned andsubjected to phylogenetic analyses with other groups ofsamples from previous mitochondrial studies [24 25 45ndash47] An NJ tree (Figure 2) of this data set gives tenuoussupport for separation into seven clusters There is goodbootstrap index (10000 replicates) for the Amerindians andAfricans haplogroups The extended sequence informationclarifies the admixture pattern evident in theGuajajaraTheseresults were consistent the sample GAJ 701 divided the

57

AMER21

AMER18AMER1962

21

12

4GAJ701

AFR50GAJ715

AMER34AMER35

AMER36GAJ718

AMER29AMER33

AMER016

A

AMER22

AMER20AMER23

62

AMER10AMER10AMER10AMER10AMER10AMER10AMER10


84

AMER31AMER32AMER3065

6073

6118

312837

1

AFR43AFR44AFR42AFR45

AFR46191067

63AMER02

GAJ717

GAJ709GAJ713

GAJ7003

4411 10

768

AFR37AFR41

AFR38AFR39AFR40

9455

60

MIT

AFR48AFR49

AFR47AMER27

AMER28AMER24AMER25

AMER26

7485

8652

465730

0002

D

C

L2

L1

L3

B

Figure 2 NJ Phylotree bootstrapped with 10000 replicates

same branch with contemporaneous Amerindian samplesthat belong to haplogroup A the samples GAJ 715 andGAJ 718 cluster with Amerindian samples according to theirrespective haplogroups D and C1 and the samples GAJ 700GAJ 709 GAJ 713 and GAJ 717 cluster with African samplesaccording to their respective haplogroups L L3e L1c and L1b

The absence of Amerindian haplogroup B in this seriesis probably due to the reduced number of examined spec-imens [13 24 27 48] However it may also reflect theunderrepresentation of this haplogroup in the formation


Table 2 Mutations assessed in the present study according to their nucleotide position their haplogroup and their respective intentionaldental modification (IDM)

Sample IDMlowastlowastlowast

Nucleotide position in the mitochondrial genome

HAPLOG

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 16 6 6 6 6 6 6 6 6 6 6 6 6 6 6 60 1 1 1 2 2 2 2 2 2 3 3 3 3 3 35 0 2 8 2 5 7 9 9 9 1 2 2 2 3 61 4 6 4 3 7 8 0 4 8 1 0 5 7 4 2

Sequence referencelowast mdash A C T C C C C C C T T C T C T T HGAJ 700 Present G mdash mdash mdash mdash T mdash mdash mdash mdash C mdash mdash mdash mdash mdash LGAJ 701 Present G mdash mdash T mdash mdash T T mdash mdash mdash mdash mdash mdash mdash C AGAJ 709 Absent G mdash mdash T T mdash T mdash mdash mdash C T mdash mdash mdash mdash L3eGAJ 713 Absent mdash mdash mdash mdash mdash T T mdash T mdash C mdash mdash mdash C mdash L1cGAJ 715 Undet mdash mdash mdash mdash mdash mdash mdash mdash mdash mdash mdash mdash C mdash mdash C DGAJ 717 Undet mdash mdash C mdash mdash mdash T mdash mdash mdash C mdash mdash mdash mdash C L1bGAJ 718 Absent mdash T mdash mdash T mdash mdash mdash mdash C C mdash C T mdash mdash C1lowastAnderson et al 1981 [35]lowastlowastlowastIDM Intentional dental modification

of the community in particular due to the mechanismsof fission-fusion [49] that rule the population growth ofindigenous groups The absence of haplogroup B in ancestralAmerindian populations is not exclusive to this study [32 50ndash53]

In the lineage classified as haplogroup A five mutationswere observed (GAJ 701 16051 16184 16278 16290and 16362) Only the mutations in bold are consensuallyaccepted asmotifs of haplogroupAThe presence of the othermutations can be justified as hotspot positions or privatemutations

Each one of the lineages classified as haplogroups C1 andD was observed in a single sample and presented six andtwo mutations respectively (GAJ 718 16104 16223 1629816311 16325 and 16327 GAJ 715 16325 and 16362) Ofthese only the mutations in bold are accepted as motifs ofhaplogroups C1 and D

Each of the lineages classified as haplogroups L L1b L1cand L3e was represented by a single sample and presentedthree four five and six mutations respectively (GAJ 70016051 16257 and 16311 GAJ 717 16126 16278 16311 and16362 GAJ 713 16257 16278 16294 16311 and 16334GAJ 709 16051 16184 16223 16278 16311 and 16320) Ofthese only themutations in bold are consensually accepted ashaplogroupmotifs In order to assign the haplogroups scien-tific articles [24 25 45 46 54ndash56] and databases specializedin mtDNA analysis HaploGrep (httphaplogrepuibkacat)and Phylotree (httpwwwphylotreeorg) were used

Analysis of mitochondrial DNA extracted from Guaja-jara ribs revealed African haplogroups in higher propor-tion (5714) than indigenous haplogroups Although thenumber of successful extractions was low this result raisesan important issue to be confirmed in future studies Thepossible penetration of African haplogroups among theGuajajara Indians suggests that women of African ancestryhave been incorporated into the Native American group

and have reproduced One cultural marker from the sameperiod as the samples from this study is the intentional dentalmodification which has never been recorded as an originalpractice of the groups of the Tupi linguistic family in Brazilandwhichwas still present among theGuajajara in themiddleof the 20th century [4]

During the period when the Guajajara bones from thisstudy originated historical documents report a decrease incontact with Europeans and a strong African and admixedpopulation presence in the Sertao of Maranhao an areatowards the East to where runaway slaves from the agri-cultural region fled to [57] and mixed with sertanejos andquilombolas who were living in an increasingly AfricanizedMaranhaoTherefore it is likely that the social circumstancesand interests among African Afrodescendant and Indige-nous groups resulted in admixture

As mentioned earlier the results obtained here are con-sistent with the data provided by Wagley and Galvao [2]Consiglio [58] Gomes [3] and other authors on theGuajajaraand their admixture with Africans The process of admixturewas possibly initiated in the 17th century and it is importantto stress that at least four slave ships arrived in Maranhaobetween 1630 and 1693 not to mention the ones that mayhave clandestinely ported on theMaranhao shoresThereforecontact and occasional admixture with Africans could havestarted very early and could have been favored by the practiceof enslaving Indigenous people who provided cheaper laborthan Africans at the time

The beginning of the slave trade in Maranhao during the17th and 18th century brought Africans from the Benin (WestAfrica) region and more recently from the Central Africancoast The presence of L1b a haplogroup mostly restricted toWest Africa [46 55 56] is consistent with our findings whichidentified four African individuals one of which belonging toL1b haplogroup


During the colonial period in Brazil both African andindigenous populations occupied the basis of the socialpyramid These groups were regarded as nothing more thanmanual laborers in an economy that was rural and slave-based This social and economic profile could be foundthroughout Brazil Slaves andAmerindians shared a commonfate which they both attempted to escape These shared lifeexperience spontaneously introduced different nonindige-nous ethnical elements into the indigenous villages whichwas also promoted by the Empirersquos strategy of mixing differ-ent ethnic groups within the coloniesThis process also led tothe appearance of the quilombos or multiethnic communitiesformed by Amerindians and Africans [46 54 57] as seen inthe present study

One of the hypotheses explaining admixture is theAfricanization of Maranhao villages after the trade activi-ties of the General Company of Grao-Para and Maranhao(Figure 1) Although reports on the first quilombos ofMaranhao date back to the end of the 19th century thereare a number of documents that address the concerns of theDiretorio dos ındios with the Guajajara due to their mixturewith ldquodeserters and fugitive slavesrdquo [3] Only two quilombolacommunities were close to the Alto Pindare region Mandidos Pretos and Buritirama [59] but there are no documentsto confirm their contact with Guajajara The first censusperformed in the state ofMaranhao in the 19th century (1872)measures in 5248 (27495238) the contribution of foreignindividuals of African origin (non-Amerindians) (IBGE)

A second wave of contact which involved admixedBrazilian people might have occurred in the Pindare areabeginning from the first half of the 19th century onwardsTheCabanos came from the state of Para between 1835 and 1841as well as the northeastern sertanejos who were fleeing fromthe severe drought in the northeastern region between 1877and 1880 These new waves of people may have introducedAfrican or people of Afrodescent to the Guajajara and wouldrevitalize villages whose populations were decreasing Thesertanejos had a significant number of women who practicedintentional dental modifications similar to those observedamong the Guajajara [11] making the sertanejos women theprobable vector of this cultural practice

Another work suggests the contact between the Africanand Amerindian populations in Brazil Gonsalves et al [60]described an unexpectedmitochondrial lineage traditionallyconsidered Polynesian among teeth samples obtained fromtwo Amerindian skulls (Botocudos who lived in the south-west Brazil at XIX century) The authors presented severalpossible scenarios to explain the admixture process amongPolynesian and Amerindian

Within this context the African haplogroups foundin this sample are of greater interest as they suggest theimportant role of African or Afrodescendent womenwho lefttheir mitochondrial evidence While the common sense con-structed around the process of admixture tends to emphasizethe role of enslaved men fleeing captivity and introducingthemselves into indigenous communities it is also importantto consider other possibilities such as the role of womenAlthough less numerous than men in the slave squadsthe presence of fugitive women is documented and proven

[57] Furthermore the hypothesis of the kidnapping andorconquest of women should also be considered Additionallythe current literature on Guajajara demography draws atten-tion to the fact that men had greater mobility while in searchof work and they probably brought back nonindigenouswomen to the villages for the purpose of marriage Thisscenario undermines the stereotype that most of the sexdynamic was between Brazilianmen thatmarried indigenouswomen It is therefore reasonable to assume that whenthe Guajajara men returned to their villages after escapingfrom oppression or captivity some of them brought alongslaves or admixed women with whom they had establisheda relationship The chronology of Guajajaras is summarizedin Box 1

Conflict of Interests

The authors declare no competing interests

Acknowledgments

This work was supported by grants from CNPq (ConselhoNacional de Desenvolvimento Cientıfico e Tecnologico ofBrazil) FINEP (Financiadora de Estudos e Projetos) CAPES(Coordenacao de Aperfeicoamento de Pessoal de NıvelSuperior) UFPA (PROPESP) Universidade Federal do Paraand FADESP (Fundacao de Amparo a Pesquisa) AndreaRibeiro-dos-Santos was supported by CNPqProdutividadeThe funders had no role in study design data collection andanalysis decision to publish or preparation of the paper

References

[1] IBGE ldquoBRASILrdquo Ministerio do Planejamento Orcamento eGestao Instituto Brasileiro de Geografia e Estatıstica 1981httpwwwibgegovbr

[2] C Wagley and E Galvao The Tenetehara Indians of Brazil ACulture in Transition Columbia University Press New YorkNY USA 1949

[3] M P GomesO Indio naHistoriamdashOPovo Tenetehara em Buscada Liberdade Vozes Rio de Janeiro Brazil 2002

[4] P E Lima ldquoDeformacoes tegumentares e mutilacao dentariaentre os ındios Teneteharardquo Boletim do Museu NacionalAntropologia vol 16 pp 1ndash22 1954

[5] M C M Alvim and J C O Gomes ldquoHiperostose porosaanemiamalarica Indios guajajaramdashestudo de casordquo inPaleopa-tologia and Paleoepidemiologia Estudos Interdisciplinares A JG Araujo and L F Ferreira Eds vol 1 of Serie PANORAMApp 141ndash158 ENSP Rio de Janeiro Brazil 1992

[6] M LocksHipoplasias Lineares de Esmalte em Indios Tenetehara-Guajajara do Estado do Maranhao Brasil Monografia deEspecializacao ENSP Rio de Janeiro Brazil 1995

[7] C Rodrigues Patologias Dento-Maxilares EmColecoes de IndiosGuajajara Monografia de Especializacao ENSP Rio de JaneiroBrazil 1995

[8] E S Cunha ldquoMutilacoes dentarias noNegro noBrasilrdquo inAnaisda Sexta Jornada Fluminense de Odontologia pp 19ndash26 Rio deJaneiro Brazil 1968


[9] W Nesi ldquoMutilacao dentaria em silvıculas de RoraimardquoArquivos Fluminenses de Odontologia vol 1 no 3 pp 8ndash141968

[10] W Nesi and V Queiroz ldquoMutilacoes dentariasrdquo in Anais daSexta Jornada Fluminense de Odontologia pp 67ndash70 Rio deJaneiro Brazil 1968

[11] L Netto Le Museum National de Rio-de-Janeiro et son Influencesur les Sciences Naturelles au Bresil Librairie C Delagrave ParisFrance 1889

[12] A Lyrio S M de Souza and C Rodrigues ldquoModificacoesdentarias na primeira catedral do Brasil Salvador BahiardquoRevista de Antropologia Portuguesa vol 18 pp 119ndash141 2001

[13] A Torroni T G Schurr C-C Yang et al ldquoNative Americanmitochondrial DNA analysis indicates that the Amerind andthe Nadene populations were founded by two independentmigrationsrdquo Genetics vol 130 no 1 pp 153ndash162 1992

[14] A Torroni R I Sukernik T G Schurr et al ldquomtDNA variationof aboriginal Siberians reveals distinct genetic affinities withNative Americansrdquo The American Journal of Human Geneticsvol 53 no 3 pp 591ndash608 1993

[15] A Torroni K Huoponen P Francalacci et al ldquoClassificationof European mtDNAs from an analysis of three Europeanpopulationsrdquo Genetics vol 144 no 4 pp 1835ndash1850 1996

[16] Y-S Chen A Torroni L Excoffier A S Santachiara-Benerecetti and D C Wallace ldquoAnalysis of mtDNA varia-tion in African populations reveals the most ancient of allhuman continent-specific haplogroupsrdquo The American Journalof Human Genetics vol 57 no 1 pp 133ndash149 1995

[17] M Richards H Corte-Real P Forster et al ldquoPaleolithic andneolithic lineages in the European mitochondrial gene poolrdquoTheAmerican Journal of Human Genetics vol 59 no 1 pp 185ndash203 1996

[18] E Watson P Forster M Richards and H-J Bandelt ldquoMito-chondrial footprints of human expansions in Africardquo TheAmerican Journal of Human Genetics vol 61 no 3 pp 691ndash7041997

[19] T Kivisild M J Bamshad K Kaldma et al ldquoDeep commonancestry of Indian and western-Eurasian mitochondrial DNAlineagesrdquo Current Biology vol 9 no 22 pp 1331ndash1334 1999

[20] VMacaulayM Richards E Hickey et al ldquoThe emerging tree ofwest EurasianmtDNAs a synthesis of control-region sequencesand RFLPsrdquo The American Journal of Human Genetics vol 64no 1 pp 232ndash249 1999

[21] D C Wallace K Garrison and W C Knowler ldquoDramaticfounder effects in Amerindianmitochondrial DNAsrdquoAmericanJournal of Physical Anthropology vol 68 no 2 pp 149ndash155 1985

[22] R H Ward B L Frazier K Dew-Jager and S Pabo ldquoExtensivemitochondrial diversity within a single Amerindian triberdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 88 no 19 pp 8720ndash8724 1991

[23] S Horai R Kondo Y Nakagawa-Hattori S Hayashi S Sonodaand K Tajima ldquoPeopling of the Americas founded by fourmajor lineages of mitochondrial DNArdquo Molecular Biology andEvolution vol 10 no 1 pp 23ndash47 1993

[24] S E B Santos A K C Ribeibo-Dos-Santos D Meyer and MA Zago ldquoMultiple founder haplotypes of mitochondrial DNAin Amerindians revealed by RFLP and sequencingrdquo Annals ofHuman Genetics vol 60 no 4 pp 305ndash319 1996

[25] A K C Ribeiro-dos-Santos S E B Santos A L Machado VGuapindaia and M A Zago ldquoHeterogeneity of mitochondrialDNA haplotypes in Pre-Columbian Natives of the Amazon

regionrdquo American Journal of Physical Anthropology vol 101 no1 pp 29ndash37 1996

[26] D G Smith R S Malhi J Eshleman J G Lorenz and F AKaestle ldquoDistribution of mtDNA haplogroup X among NativeNorth Americansrdquo American Journal of Physical Anthropologyvol 110 no 3 pp 271ndash284 1999

[27] M Moraga C M Santoro V G Standen P Carvallo and FRothhammer ldquoMicroevolution in prehistoric Andean popula-tions chronologic mtDNA variation in the desert valleys ofnorthernChilerdquoAmerican Journal of Physical Anthropology vol127 no 2 pp 170ndash181 2005

[28] P Forster R Harding A Torroni and H-J Bandelt ldquoOriginand evolution of native American mtDNA variation a reap-praisalrdquo The American Journal of Human Genetics vol 59 no4 pp 935ndash945 1996

[29] P Forster ldquoIce Ages and the mitochondrial DNA chronologyof human dispersals a reviewrdquo Philosophical Transactions of theRoyal Society B Biological Sciences vol 359 no 1442 pp 255ndash264 2004

[30] J C Rando F Pinto A M Gonzalez et al ldquoMitochondrialDNA analysis of Northwest African populations reveals geneticexchanges with European Near-Eastern and sub-Saharan pop-ulationsrdquo Annals of Human Genetics vol 62 no 6 pp 531ndash5501998

[31] Y-S Chen A Olckers T G Schurr A M Kogelnik KHuoponen and D C Wallace ldquomtDNA variation in the SouthAfrican Kung and Khwemdashand their genetic relationships toother African populationsrdquo The American Journal of HumanGenetics vol 66 no 4 pp 1362ndash1383 2000

[32] A N D R Marinho N C Miranda V Braz A K Ribeiro-dos-Santos and S M F M de Souza ldquoPaleogenetic andtaphonomic analysis of human bones from Moa Beirada andZe Espinho Sambaquis Rio de Janeiro Brazilrdquo Memorias doInstituto Oswaldo Cruz vol 101 supplement 2 pp 15ndash23 2006

[33] J Sambrook E F Fritschi andTManiatisMolecular Cloning ALaboratoryManual Cold SpringHarbor Laboratory Press NewYork NY USA 1989

[34] F Sanger S Nichelen and A R Coulson ldquoDNA sequencingwith chain-terminating inhibitorsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 74 no12 pp 5463ndash5468 1977

[35] S Anderson A T Bankier A G Barriel et al ldquoSequence andorganization of the humanmitochondrial genomerdquoNature vol290 no 5806 pp 457ndash465 1981

[36] N Saitou and M Nei ldquoThe neighbor-joining method a newmethod for reconstructing phylogenetic treesrdquo Molecular Biol-ogy and Evolution vol 4 no 4 pp 406ndash425 1987

[37] H-J Bandelt P Forster B C Sykes and M B RichardsldquoMitochondrial portraits of human populations using mediannetworksrdquo Genetics vol 141 no 2 pp 743ndash753 1995

[38] H-J Bandelt P Forster andA Rohl ldquoMedian-joining networksfor inferring intraspecific phylogeniesrdquo Molecular Biology andEvolution vol 16 no 1 pp 37ndash48 1999

[39] H-J Bandelt VMacaulay andM Richards ldquoMedian networksspeedy construction and greedy reduction one simulation andtwo case studies from humanmtDNArdquoMolecular Phylogeneticsand Evolution vol 16 no 1 pp 8ndash28 2000

[40] A Cooper and H N Poinar ldquoAncient DNA do it right or notat allrdquo Science vol 289 no 5482 p 1139 2000

[41] M T P Gilbert H-J Bandelt M Hofreiter and I BarnesldquoAssessing ancient DNA studiesrdquo Trends in Ecology amp Evolutionvol 20 no 10 pp 541ndash544 2005


[42] M Hofreiter D Serre H N Poinar M Kuch and S PaaboldquoAncient DNArdquo Nature Reviews Genetics vol 2 no 5 pp 353ndash359 2001

[43] S Paabo H Poinar D Serre et al ldquoGenetic analyses fromancient DNArdquo Annual Review of Genetics vol 38 pp 645ndash6792004

[44] H-J Bandelt L Quintana-Murci A Salas and V MacaulayldquoThe fingerprint of phantom mutations in mitochondrial DNAdatardquo The American Journal of Human Genetics vol 71 no 5pp 1150ndash1160 2002

[45] A C Feio-dos-Santos B M Carvalho S E B dos Santos andA K C Ribeiro-dos-Santos ldquoNucleotide variability of HV-I inadmixed population of the Brazilian Amazon Regionrdquo ForensicScience International vol 164 no 2-3 pp 276ndash277 2006

[46] B M Carvalho M C Bortolini S E B dos Santos and AK C Ribeiro-dos-Santos ldquoMitochondrial DNA mapping ofsocial-biological interactions in Brazilian Amazonian African-descendant populationsrdquo Genetics and Molecular Biology vol31 no 1 pp 12ndash22 2008

[47] P Taboada-Echalar V Alvarez-Iglesias T Heinz et al ldquoThegenetic legacy of the pre-colonial period in contemporaryBoliviansrdquo PLoS ONE vol 8 no 3 Article ID e58980 2013

[48] J Garcıa-Bour A Perez-Perez E Prats andD Turbon ldquoMolec-ular approach to the peopling of the Americas by sequencingmtDNA from extinct Fueguians and Patagonsrdquo Chungara vol32 no 2 pp 265ndash266 2000

[49] J V Neel and F M Salzano ldquoFurther studies on the XavanteIndians X Some hypotheses-generalizations resulting fromthese studiesrdquoTheAmerican Journal of Human Genetics vol 19no 4 pp 554ndash574 1967

[50] P K Rogan and J J Salvo ldquoMolecular genetics of Pre-Columbi-an South American mummiesrdquoUCLA Symposium inMolecularEvolution vol 122 pp 223ndash234 1990

[51] S Horai R Kondo K Murayama S Hayashi H Koike andN Nakai ldquoPhylogenetic affiliation of ancient and contempo-rary humans inferred from mitochondrial DNArdquo PhilosophicalTransactions of the Royal Society B Biological Sciences vol 333no 1268 pp 409ndash416 1991

[52] D A Merriwether F Rothhammer and R E Ferrell ldquoGeneticvariation in the New World ancient teeth bone and tissue assources of DNArdquo Experientia vol 50 no 6 pp 592ndash601 1994

[53] D A Demarchi G M Panzetta-Dutari S E Colantonio andA J Marcellino ldquoAbsence of the 9-bp deletion of mitochondrialDNA in pre-Hispanic inhabitants of Argentinardquo Human Biol-ogy vol 73 no 4 pp 575ndash582 2001

[54] A K C Ribeiro-dos-Santos J M PereiraM R F Lobato BMCarvalho J F Guerreiro and S E B dos Santos ldquoDissimilaritiesin the process of formation of Curiau a semi-isolated Afro-Brazilian population of the Amazon regionrdquo American Journalof Human Biology vol 14 no 4 pp 440ndash447 2002

[55] A Salas M Richards T de la Fe et al ldquoThe making of theAfrican mtDNA landscaperdquo The American Journal of HumanGenetics vol 71 no 5 pp 1082ndash1111 2002

[56] A Salas M Richards M-V Lareu et al ldquoThe African diasporamitochondrial DNA and the Atlantic slave traderdquoTheAmericanJournal of Human Genetics vol 74 no 3 pp 454ndash465 2004

[57] M S Amantino O Mundo das Feras Os Moradores do SertaoOeste de Minas GeraismdashSeculo XVIII vol 1 UFRJIFCS Rio deJaneiro Brazil 2001

[58] V Consiglio Fontes Missionarias e Historia Indıgena Uminventario analıtico sobre textos jesuıticos nos arquivos romanos

referentes a missao em Maranhao e Grao-Paramdashseculos XVII-XVIII USP Sao Paulo Brazil 1997

[59] R S A Anjos andACiprianoQuilombolas Tradicoes e Culturada Resistencia Aori Comunicacao Sao Paulo Brazil 2006

[60] V F Goncalves J Stenderup C Rodrigues-Carvalho et alldquoIdentification of PolynesianmtDNAhaplogroups in remains ofBotocudoAmerindians fromBrazilrdquoProceedings of theNationalAcademy of Sciences of the United States of America vol 110 no16 pp 6465ndash6469 2013

Submit your manuscripts athttpwwwhindawicom

Child Development Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Education Research International


Biomedical EducationJournal of



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ArchaeologyJournal of



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Research and TreatmentSchizophrenia


Urban Studies Research

Population ResearchInternational Journal of


CriminologyJournal of


Aging ResearchJournal of



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Current Gerontologyamp Geriatrics Research

Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

Sleep DisordersHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

AddictionJournal of


Depression Research and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Geography Journal


Research and TreatmentAutism


Economics Research International


48∘09984000998400998400W

47∘09984000998400998400W

46∘09984000998400998400W

45∘09984000998400998400W

44∘09984000998400998400W

43∘09984000998400998400W

42∘09984000998400998400W

1∘09984000998400998400S

2∘09984000998400998400S

3∘09984000998400998400S

4∘09984000998400998400S

5∘09984000998400998400S

6∘09984000998400998400S

7∘09984000998400998400S

8∘09984000998400998400S

9∘09984000998400998400S

10∘09984000998400998400S

Imperatriz

Guajajara

Quilombo

tribes

populationsSatildeo Luiacutes

Figure 1 Current distribution of the studied Guajajara villages ofKamiranga and Januaria (in red) and the quilombola populations(in black) in the state of Maranhao Brazil (adapted from [59])

Galvao [2] and Gomes [3] the Guajajararsquos traditional kinshipsystem is flexible enough to accommodate the need for seek-ing marriages in more distant and less related communitiesthereby acting as a buffer during the most critical demo-graphic periods The historical conditions of admixture withother ethnic groups throughout four centuries of contacts aremapped in the historical-anthropological revision by Gomes[3] However the precise time that the first events of contactwith African andor Afrodescendants started is not clear inthe historical data (Box 1)

12 Bioanthropology and Bioarchaeology of Guajajara TheGuajajara are currently one of the most documented ethnicgroups of Brazil Many studies have recorded the ethnog-raphy demography history bioanthropology and bioar-chaeology of the Guajajara [4ndash7] Lima [4] documentedthe interesting practice of intentional dental modificationprevalent among the Guajajara and it was confirmed in someexhumed skulls The dental modification was characterizedby symmetric cuts of the lateral and medial angles of theincisors called ldquopiranha teethrdquo and was attributed to theadoption of African habits This interpretation was based onoral information about Afrodescendants from some of theGuajajara in the villages Other biological features other thansome adopted material culture music and customs of theseIndians were also recognized as being of Afro-Americanorigin In the past dental modification was found in severalregions of Africa In Brazil other indigenous groups such

as the Tucuna Makuxi Galibi Karipuna Pauaxiana Jauariand Kaigang also adopted similar dental modifications forthe same reasons since the practice of intentional dentalmodification has never been demonstrated in Brazilianarcheological sites prior to contact [4 8ndash10]This practice hasalready been described amongAfricans andAfrodescendantsin Brazil [9 11 12]

The distribution of genetic patterns of the ancestralmtDNA complements and confirms the results obtained bymorphological and archeological studies it has enabled theformulation of a more adequate model to explain prehistoricdiversity and the peopling of the Americas These char-acteristics confer significant advantages of the use of themitochondrial genome for the research in the present studyHowever one must not forget a significant limitation froma biological perspective it only narrates one part of humanhistory that of the maternal lineage

The specificity of mtDNA lineages among different popu-lations and groups has favored their use as ancestry markersover the past few decades [13ndash20] Among Amerindianpopulations five main haplogroups are prevalent (A B CD and Xmdash[21 22]) [13 23ndash27] Asian populations shareAmerican haplogroups (A B C and D) as well as otherhaplogroups (M N G F Z Y and R) that are described asspecific to the Asian continent [28 29] European and Euro-Asian populations also possess more frequent haplogroups(I J K H T U V and W) [15 28 29] In contrast Africanpopulations have a large amount of exclusive haplogroupsand subhaplogroups (L0 L1 L2 L3 L4 L5 and L6) [18 3031]

This study is a first step toward elucidating the geneticcharacteristics of the Guajajara individuals Our objective isto characterize the admixture with non-Amerindian popula-tions In addition to contributing to our understanding of thehistory of this ethnic group this study aims to corroborateprevious bioanthropological studies and to contribute tothe knowledge of regional population dynamics during thecenturies that followed colonization until the final formationof what is today the Brazilian national society The presentstudy uses skeletal samples from the Guajajara cemeteriesto investigate aDNA mitochondrial haplogroups and tocontribute to the reconstruction of part of its history

2 Materials and Methods

21 Subjects The bone samples belong to the BiologicalAnthropology collection of the National Museum Rio deJaneiro The samples were obtained from 12 different indi-viduals exhumed from cemeteries of the Guajajara villages ofJanuaria and Kamiranga (Figure 1) These skeletons are partof the collection brought by Pedro Lima and his collaboratorsin 1945

The bone samples were collected from ribs removed fromeach individual The ribs were selected following criteria ofintegrity absence of pathological signs anomalies or severetaphonomic changes that could compromise future inves-tigations about the health state of other parts of the skeletons(bioarchaeological studies such as age and sex assessment or










13

corrected)















57

AMER21

AMER18AMER1962

21

12

4GAJ701

AFR50GAJ715

AMER34AMER35

AMER36GAJ718

AMER29AMER33

AMER016

A

AMER22

AMER20AMER23

62



84


6073

6118

312837

1


AFR46191067

63AMER02

GAJ717

GAJ709GAJ713

GAJ7003

4411 10

768

AFR37AFR41

AFR38AFR39AFR40

9455

60

MIT

AFR48AFR49

AFR47AMER27

AMER28AMER24AMER25

AMER26

7485

8652

465730

0002

D

C

L2

L1

L3

B








HAPLOG

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 16 6 6 6 6 6 6 6 6 6 6 6 6 6 6 60 1 1 1 2 2 2 2 2 2 3 3 3 3 3 35 0 2 8 2 5 7 9 9 9 1 2 2 2 3 61 4 6 4 3 7 8 0 4 8 1 0 5 7 4 2




















Acknowledgments


References









































































Psychiatry Journal



















Volume 2014


AddictionJournal of




Geography Journal














13

corrected)















57

AMER21

AMER18AMER1962

21

12

4GAJ701

AFR50GAJ715

AMER34AMER35

AMER36GAJ718

AMER29AMER33

AMER016

A

AMER22

AMER20AMER23

62



84


6073

6118

312837

1


AFR46191067

63AMER02

GAJ717

GAJ709GAJ713

GAJ7003

4411 10

768

AFR37AFR41

AFR38AFR39AFR40

9455

60

MIT

AFR48AFR49

AFR47AMER27

AMER28AMER24AMER25

AMER26

7485

8652

465730

0002

D

C

L2

L1

L3

B








HAPLOG

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 16 6 6 6 6 6 6 6 6 6 6 6 6 6 6 60 1 1 1 2 2 2 2 2 2 3 3 3 3 3 35 0 2 8 2 5 7 9 9 9 1 2 2 2 3 61 4 6 4 3 7 8 0 4 8 1 0 5 7 4 2




















Acknowledgments


References









































































Psychiatry Journal



















Volume 2014


AddictionJournal of




Geography Journal













57

AMER21

AMER18AMER1962

21

12

4GAJ701

AFR50GAJ715

AMER34AMER35

AMER36GAJ718

AMER29AMER33

AMER016

A

AMER22

AMER20AMER23

62



84


6073

6118

312837

1


AFR46191067

63AMER02

GAJ717

GAJ709GAJ713

GAJ7003

4411 10

768

AFR37AFR41

AFR38AFR39AFR40

9455

60

MIT

AFR48AFR49

AFR47AMER27

AMER28AMER24AMER25

AMER26

7485

8652

465730

0002

D

C

L2

L1

L3

B








HAPLOG

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 16 6 6 6 6 6 6 6 6 6 6 6 6 6 6 60 1 1 1 2 2 2 2 2 2 3 3 3 3 3 35 0 2 8 2 5 7 9 9 9 1 2 2 2 3 61 4 6 4 3 7 8 0 4 8 1 0 5 7 4 2




















Acknowledgments


References









































































Psychiatry Journal



















Volume 2014


AddictionJournal of




Geography Journal









HAPLOG

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 16 6 6 6 6 6 6 6 6 6 6 6 6 6 6 60 1 1 1 2 2 2 2 2 2 3 3 3 3 3 35 0 2 8 2 5 7 9 9 9 1 2 2 2 3 61 4 6 4 3 7 8 0 4 8 1 0 5 7 4 2




















Acknowledgments


References









































































Psychiatry Journal



















Volume 2014


AddictionJournal of




Geography Journal














Acknowledgments


References









































































Psychiatry Journal



















Volume 2014


AddictionJournal of




Geography Journal





































































Psychiatry Journal



















Volume 2014


AddictionJournal of




Geography Journal





research article paleogenetic studies in guajajara

Documents