A paleodietary Study of Stable Isotope Analysis from a High-status Burial in the Copper Age: The Montelirio Megaithic Structure at Valencina de la Cncepción-Castilleja de Guzmán,

A Palaeodietary Study of Stable IsotopeAnalysis from a High-status Burial in theCopper Age: The Montelirio MegalithicStructure at Valencina de la Concepción–Castilleja de Guzmán, SpainM. FONTANALS-COLL,a* M. DÍAZ-ZORITA BONILLAb AND M. E. SUBIRÀa

a GRAPAC Grup de Recerca Aplicada al Patrimoni Cultural, Unitat d’Antropologia Biològica, Departament deBiologia Animal, de Biologia Vegetal i d’Ecologia, Universitat Autònoma de Barcelona, Barcelona, Spainb Department of Geosciences, Biogeologie, University of Tübingen, Tübingen, Germany

ABSTRACT This palaeodietary study presents carbon and nitrogen stable isotope data from human and faunal skeletalremains from the Copper Age settlement of Valencina de la Concepción–Castilleja de Guzmán, located inSeville, Spain. Montelirio, the only Valencina–Castilleja human group from which we have obtained reliablepalaeodietary results, had a diet based on C3 terrestrial resources, including both plants and animals. Theprotein component of the diet consisted mainly of meat, milk and dairy products from livestock as well asC3 plant protein from cereals and pulses. This study compares data from Montelirio, the Copper Age groupfrom Valencina–Castilleja, with the published data from other Iberian Late Neolithic–Copper Age (LN-CA) so-cieties. This comparison reveals a homogeneous diet with some exceptions. Overall, the LN-CA diet in theIberian Peninsula consisted mainly of animal proteins from meat, milk and dairy products from livestock aswell as C3 plant sources from the characteristic agriculture of the societies of these periods. This study alsodemonstrates the minor use of aquatic resources from the Neolithic to the Copper Age period in the IberianPeninsula. Copyright © 2015 John Wiley & Sons, Ltd.

Key words: carbon; Copper Age; nitrogen; palaeodiet; stable isotopes; Valencina de la Concepción–Castilleja de Guzmán

Introduction

The reconstruction of prehistoric diets through stablecarbon and nitrogen isotope analysis from bone colla-gen has already been active for several years. However,because of difficulties with collagen preservation, thesestudies are not always easy, particularly in hot climates,such as those of southern European countries.Until now, few isotopic analyses have been per-

formed on the Iberian Peninsula. Most have focusedon understanding prehistoric populations from theMesolithic (e.g. Garcia-Guixé et al., 2006; Arias andSchulting, 2010; Fernández-López de Pablo et al.,2013; Fontanals-Coll et al., 2014) through the

Neolithic (e.g. Lubell et al., 1994) and Copper Age(Díaz-Zorita Bonilla, 2013) up to the Iron Age (e.g.Fuller et al., 2010; Salazar-García, 2011). Isotopicstudies in the southern area of the Iberian Peninsulaare rarer.The present isotopic study analyses data from hu-

man and faunal skeletal remains from the Copper Agesettlement of Valencina de la Concepción–Castillejade Guzmán (henceforth ‘Valencina–Castilleja’), Seville,Spain (Figure 1).The Iberian Copper Age spanned from ca.

3500–2250cal BC (Díaz del Río, 2004). This period isidentified mainly by the increase in the number andsize of sites, higher population densities andagricultural intensification (García-Sanjuán andMurillo-Barroso, 2012). However, these patterns havevaried throughout the Iberian Peninsula, and regionaldifferences exist (for detailed information on each area,see Gibaja et al., 2012). Burials in megalithic

* Correspondence to: Maria Fontanals-Coll, Unitat d’AntropologiaBiològica, Departament de Biologia Animal, Biologia Vegetal i Ecologia(BABVE), Facultat de Biociències (Edifici C), Universitat Autònoma deBarcelona (UAB), 08193 Cerdanyola del Vallès, Barcelona, Spain.e-mail: [email protected]

Copyright © 2015 John Wiley & Sons, Ltd. Received 30 January 2014Revised 3 December 2014

Accepted 23 December 2014

International Journal of OsteoarchaeologyInt. J. Osteoarchaeol. (2015)Published online in Wiley Online Library(wileyonlinelibrary.com) DOI: 10.1002/oa.2435

monuments were still prevalent in this period, althoughother structures such as pits and natural caves were alsoin use. The burial rite during the third millennium BCseems to be mainly collective; inhumation is predomi-nant, but cremations have also been documented(Agustí, 2002; Silva et al., 2012).During this period, evidence exists of long-distance

networks, craft specialisation and the use of certainraw materials as prestige objects in the funerary con-text. One of the best-studied Copper Age sites thatinclude all these indicators is Valencina–Castilleja,which is located in Seville, in the south-west of theIberian Peninsula.The Copper Age settlement of Valencina–Castilleja

is located 6km from the city of Seville, in south-westSpain. It is situated in the Aljarafe plateau, west ofthe Guadalquivir River, and it covers approximately400ha. This area is very rich in natural resources,and the ground is fertile. Freshwater resources areavailable because it was located next to marine andlake environments and near important forestry re-sources. According to the few published studies aboutthe site, the main two activities were cultivation andlivestock management in which cattle and suids werethe main animals exploited (e.g. Pajuelo and López,2013; Martínez, 2013). These results are consistentwith the exploitation of the landscape since the Neo-lithic period, based on the so-called ‘policultivoganadero’, an extensive pastoralist strategy (Harrison,

1985) along with dry forest management practices(a mixed exploitation of forest resources and livestockfarming). The modification of the landscape washighly visible during Late Prehistory in Iberia fromthe fourth millennium BC to the second millenniumAD (Stevenson and Harrison, 1992).Valencina–Castilleja was first investigated at the end

of the 19th century based on the study of the mega-lithic monuments of La Pastora, Matarrubilla andOntiveros (e.g. Carriazo, 1962; Collantes de Terán,1969). However, investigations have continued sincethen (for a complete review, see Vargas, 2004; Costaet al., 2010; García-Sanjuán, 2013).This site thus represents one of the most inten-

sively excavated Copper Age settlements in Iberia,and many different structures and monuments havebeen identified. The radiocarbon dates suggest thatsettlement at Valencina–Castilleja has a chronologi-cal range from the beginning of the third millennium(3000–2800 cal BC) until the first half of the secondmillennium BC (1710–1404 cal BC 2σ) (Noceteet al., 2011; García-Sanjuán, 2013).In 1998, a major megalithic monument, Montelirio,

similar to La Pastora and Matarrubilla, was discovered(Férnadez and Aycart, 2013). Montelirio representsone of the most recent excavations documented inValencina–Castilleja. The monument is a tholos witha double chamber, oriented on the east–west axis(Fernández and Aycart, 2013). Two individuals were

Figure 1. Location of Valencina–Castilleja, Seville (Spain), and the Late Neolithic–Copper Age sites from the Iberian Peninsula used for the study.

M. Fontanals-Coll et al.

Copyright © 2015 John Wiley & Sons, Ltd. Int. J. Osteoarchaeol. (2015)

found buried in the corridor, and the rest (minimumnumber of individuals=24) were found in the mainchamber (Pecero et al., 2012). The funerary ritual ofthe main chamber was collective, and the burials inboth the corridor and main chamber were found tobe associated with a great diversity of grave goods, in-cluding ceramics, lithics, shells and metals that werefound along with exotic goods such as amber, ivoryand ostrich egg shells. Similar findings have previouslybeen documented at Valencina–Castilleja. For exam-ple, flint and flint objects (e.g. Morgado et al., 2011),ivory (e.g. Schuhmacher et al., 2012), amber (e.g.Murillo-Barroso and Martinón-Torres, 2012) and greenstones (Odriozola et al., 2010) have been found. Thehigh-status burial of Montelirio represents a good ex-ample of a specific segment of the society withexcellent preservation. Montelirio represents an excep-tional burial among the prehistoric monuments ofValencina–Castilleja and also within the Late Prehis-tory of the Spanish south-west. The excellent preserva-tion of the osteological record, the biological remainsand the burial materials make this site unique. In addi-tion, the funerary structure (one of the few large mega-lithic structures) itself is another element of socialdifferentiation. The different chronological phasesand the evolution of this deposit are explained in detailby Fernández and Aycart (2013). This monument rep-resents one of the few well-preserved Copper Ageburials in Iberia that contain high-status objects, in-cluding some allochthonous objects. Therefore, thissite is very significant for the study of social complexityand social differences during Late Prehistory in Iberia.The other studied sector from Valencina–Castilleja

is a surrounding area of the megalithic monument ofLa Pastora. Many negative structures with several pitsand a ditch were found in this sector (Vargas Jiménez,2013), including a circular structure where the individ-ual under study was found.This paper aims to analyse the diet and the subsis-

tence economy of the human individuals found in thehigh-status burial of Montelirio and the La Pastoracomplex from Valencina–Castilleja, by stable carbon

and nitrogen isotope analysis of human and faunalskeletal remains. To highlight similarities and differencesin diet between Late Neolithic and Copper Agesocieties, the Valencina–Castilleja results were comparedwith the published analyses of other contemporary sitesfrom the Iberian Peninsula (García et al., 2010; McClureet al., 2011;Waterman, 2012;Waterman et al., 2014). It isimportant to highlight that the megalithic constructionsreveal continuity between Portuguese and Spanish sites,which reinforces the validity of this comparison. ThePortuguese sites present a long occupational period thatincludes the Copper Age.

Materials and methods

The isotopical analysis performed in this study presentsdata from human and faunal remains collected in theMontelirio megalithic monument and from the LaPastora complex at the Valencina–Castilleja site. TheMontelirio sector dates from 3000 to 2200cal BC, ac-cording to the radiocarbon dating of the funerary con-text (Table 1).The remains include bone material from 15 humans

and five faunal bone samples of Sus sp., Ovis/Capra sp.and Bos taurus from Montelirio and material from onehuman and five faunal samples of Equus sp., Capra sp.,Cervus sp., Sus sp. and B. taurus from La Pastora.The age and sex diagnosis and the osteological

methods used are explained in detail by Pecero et al.(2012). The sampling process was conducted byselecting long bone fragments from lower extremities.All human and faunal individuals available for stableisotope analysis were sampled.The stable carbon and nitrogen isotope analyses

from bone collagen provide information about the die-tary content of the individuals, based on the protein in-take. In this sense, the stable carbon isotope value(δ13C) distinguishes the consumption of marine (13C-enriched) versus terrestrial C3-based foods, with theo-retical endpoints of approximately �21‰ to �20‰for a major terrestrial protein intake and approximately

Table 1. Radiocarbon-calibrated ages from Montelirio sector

Site Laboratory code Context 14C age BP14C age BC (calibrated 2σ) Source

Montelirio Ua-40804 Funerary 3862± 30 2470–2200 Fernández and Aycart, 2013Ua-40802 Funerary 4002± 31 2580–2460Ua-40805 Funerary 4086± 35 2862–2494CNA-588 Funerary 4060± 40 2851–2476CNA-585 Funerary 4250± 35 2923–2698Ua-40801 Funerary 4180± 30 2890–2660Ua-40803 Funerary 4165± 30 2880–2630CNA-586 Funerary 4140± 50 2880–2575

A Palaeodietary Study of a Copper Age Burial, Valencina, Spain


�12‰ for a purely marine protein consumption. Thisvalue also helps to discriminate between the input ofC3 (13C-depleted) and C4 (13C-enriched) plant sourcesinto the diet (Schoeninger and DeNiro, 1984;Schwarcz and Schoeninger, 1991; Richards, 2002;Lillie, 2003; Eriksson et al., 2008, Lillie, et al., 2011).However, when the consumption of both C4 plantsand marine resources is present in the individual’s diet,there could appear a difficulty in the interpretation. Butthis complication does not arise in this study because ofthe insignificant quantities of edible C4 plants duringthe Chalcolithic in Southern Iberia. The stable nitro-gen isotope value (δ15N) provides information aboutthe trophic level of each individual: the values of con-sumers present an increase of 3–5‰ over the valuesof their diets in the food chain (De Niro and Epstein,1981; Schoeninger and De Niro, 1984; Bocherens andDrucker, 2003; Richards et al., 2005; Lillie et al., 2011).However, it is always important to take into account thedifficulty of distinguishing animal-rich diets from plant-rich diets using δ15N values discussed in some studies suchas Hedges and Reynard (2007) and Fraser et al. (2013).The stable isotope signature of an adult human bone

reflects the average diet of at least the last 10–15yearsof life because of the rate of the bone turnover duringthe individual’s lifetime (Hedges et al., 2007).Established quality indicators are used to ensure thepreservation quality of the extracted collagen(De Niro, 1985; Ambrose, 1993; Van Klinken, 1999).The collagen extraction process for this study was per-

formed at the laboratories of the Unitat d’AntropologiaBiològica of theUniversitat Autònoma de Barcelona. Priorto analysis, mechanical abrasion with a multifunction toolDremel 3000 (Dremel Europe), with a diamond engravingtip, was used to remove visible contaminants. Collagenextraction proceeded following Richards and Hedges(1999), including Brown et al. (1988) and modificationsreferring to the ultrafiltration step. The measurementsfor the stable isotope analysis were performed at theInstitut de Ciència i Teconologia Ambientals using aThermo Flash 1112 (Thermo Scientific) elementalanalyser coupled to a Thermo Delta V Advantage massspectrometer (Thermo Scientific) with a Conflo III inter-face, which measures the ratios of 13C to 12C and 15Nto 14N relative to a standard (Vienna Pee Dee Belemnite(V-PDB) for carbon and air for nitrogen) and expressesthe stable isotope values in parts per thousand (‰) andin delta notation (δ). The international laboratory stan-dard, IAEA 600 (caffeine), is used here. An analyticalerror always below 0.2‰ (1σ) in all the repeated analyseswas determined for δ13C and δ15N. The descriptive statis-tics were calculated using the IBM Statistics Software SPSS20.0.0 (Armonk, NY, USA). Ta

ble

2.Stable

carbon

(δ13C)an

dnitrog

en(δ

15N)va

lues

obtained

from

faun

alsa

mplesfrom

Valen

cina

–Cas

tilleja:M

ontelirio

andLa

Pas

tora,ind

icatingco

llagen

control

indicators(yield,%

C,%

Nan

dC:N

)an

dsa

mpledbon

e

Site

Figures

code

Laboratory

code

Individua

lTa

xaδ1

3C

δ15N

Yield

(%)

%C

%N

C:N

Sam

pledbon

e

Mon

telirio

12MoF17

UE-253

Sus

sp.

�19.6

4.2

1.3

40.9

14.4

3.3

Pha

lanx

13MoF18

UE-253

Ovis/Cap

rasp

.�1

9.4

5.6

1.1

41.6

14.7

3.3

Long

bon

ediaphy

sis

14MoF19

UE-235

Ovis/Cap

rasp

.�1

8.7

5.6

0.9

45.3

16.1

3.3

Long

bon

ediaphy

sis

—MoF20

.1UE-17Ind.1

Bos

taurus

——

——

——

Lefttib

ia—

MoF20

.2UE-17Ind.2

Ovis/Cap

rasp

.—

——

——

—Lo

ngbon

ediaphy

sis

La Pas

tora

—Pa1

UC54

UD72

(63)

Equu

ssp

.—

——

——

—Lo

ngbon

ediaphy

sis

—Pa4

UC20

6UD20

7(101

)Cervu

ssp

.�2

0.8

4.7

0.8

35.1

12.7

2.8a

Long

bon

ediaphy

sis

15Pa5

UC54

UD72

(13)

Cap

rasp

.�2

0.4

5.9

1.1

39.8

13.7

2.9

Long

bon

ediaphy

sis

16Pa6

UC20

6UD23

4(23)

Bos

taurus

�18.7

6.2

1.4

38.9

12.7

2.9

Leftfemur

—Pa7

UC20

6UD23

4(36)

Sus

sp.

�20.6

9.0

0.5

38.9

14.1

2.8a

Long

bon

ediaphy

sis

aValue

sfalling

outsidetheac

ceptedrang

eforgoo

dco

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prese

rvation.



Results

The isotope data obtained from the human and faunalskeletal material from Montelirio and La Pastora sectorsat Valencina–Castilleja are presented in Tables 2 and 3and Figure 2.Only three human samples, Mo1 (UE-251), Mo7

(UE-229) (the only male individual from Montelirio)and Mo9 (UE-232) of a total of 15 human remainsdid not yield enough collagen to run samples in dupli-cate. One human sample Mo12 (UE-115) and threesamples from La Pastora [the two faunal samples, Pa4(UC206 UD207 101) Cervus sp. and Pa7 (UC206

UD234 36) Sus sp., and the single human sample iden-tified as Pa3] did not meet the published quality criteria(De Niro, 1985; Ambrose 1993; Van Klinken, 1999)and are excluded from all the summary calculations.Geographical variation in local isotopic baselines has

been demonstrated by Van Klinken et al. (1994),Hedges et al. (2004) and Fischer et al. (2007). For thisstudy, we calculated endpoints based on measurementsof local fauna using offsets of approximately 1‰ forδ13C and 3.5‰ for δ15N (e.g. Minagawa and Wada,1984; Schoeninger and De Niro, 1984; Richards andHedges, 1999; Bocherens and Drucker, 2003; Fischeret al., 2007). The δ13C values of the herbivores[�19.3‰±0.8 (1σ)‰] and the single omnivore(�19.6‰) are consistent with the values for a terres-trial C3 European ecosystem (De Niro and Epstein,1978; Schwarcz and Schoeninger, 1991). And theδ15N values are 5.8‰±0.3 (1σ)‰ and 4.2‰, respec-tively. To define the local trophic baseline for the hu-man palaeodietary reconstruction, we used only theherbivore mean value for calculations.By combining the faunal data, a theoretical terrestrial

endpoint of approximately �18.3‰ (approximately�19.3‰, adjusted 1‰) can be suggested from themean δ13C value of terrestrial herbivores. The marineendpoint estimation of �12±1‰ was used(Francalacci, 1989; Jennings et al., 1997; Richards andHedges, 1999). The theoretical endpoint or terrestrialmammal protein consumption was calculated fromthe mean δ15N value ~5.8‰ and, adjusted 3–5‰for the trophic level shift correction, set a range of8.8–10.8‰ (9–11‰, after rounding) for an animalprotein intake in the human diet.

Table 3. Stable carbon (δ13C) and nitrogen (δ15N) values obtained from human samples from Valencina–Castilleja: Montelirio and LaPastora, indicating collagen control indicators (yield, %C, %N and C :N) and sampled bone

SiteFigurescode

Laboratorycode Individual

Age(years) Sex δ13C δ15N

Yield(%) %C %N C :N Sampled bone

Montelirio ― Mo1 UE-251 20–30 Female? ― ― ― ― ― ― Right femur1 Mo2 UE-101 25–30 Female �18.3 8.7 1.2 30.3 10.3 3.4 Right tibia2 Mo3 UE-102 25–34 Female �19.1 9.0 1.9 36.3 12.7 3.3 Right femur3 Mo4 UE-103 20–30 Female �19.1 8.9 2.1 36.6 12.9 3.3 Right tibia4 Mo5 UE-114 18–30 Female �19.9 9.1 1.7 32.3 11.4 3.3 Left femur5 Mo6 UE-112 30–35 Female �19.4 10.0 1.4 32.1 11.4 3.3 Right tibia

― Mo7 UE-229 Adult Male ― ― ― ― ― — Right humerus― Mo9 UE-232 Adult Indet. ― ― ― ― ― — Right tibia6 Mo10 UE-370 Adult Indet. �20.3 8.9 1.0 30.9 10.6 3.4 Right tibia7 Mo11 UE-107 30–39 Female �19.5 9.5 0.9 35.1 12.2 3.3 Right femur

― Mo12 UE-115 Adult Indet. �23.7 10.0 0.4 13.9a 4.4a 3.7a Left femur8 Mo13 UE-346 20–30 Female? �20.0 8.9 1.1 36.4 12.9 3.3 Right tibia9 Mo14 UE-343 24–35 Female �19.3 8.9 2.2 43.0 14.8 3.4 Pelvis

10 Mo15 UE-360 18–30 Female �19.3 9.0 1.6 35.4 12.4 3.3 Femur11 Mo16 UE-111 20–30 Female �20.4 9.2 1.7 32.4 11.4 3.3 Right femur

La Pastora ― Pa3 ― Adult Indet. �19.5 8.6 0.7 35.2 12.8 2.8a Right tibia

aValues that fall outside the accepted range for good collagen preservation.

Figure 2. Plot of δ13C and δ15N values, expressed in parts per thou-sand, of bone collagen from human and fauna individuals from theValencina–Castilleja sites. The numbers correspond to the code of eachindividual, which can be observed in Tables 1 and 2.



The mean δ13C value of the human group fromValencina–Castilleja is �19.5±0.6 (1σ)‰, and themean δ15N value is 9.1±0.4 (1σ)‰. These values areconsistent with a diet based mainly on C3 terrestrial re-sources and place all humans in a higher trophic levelthan the herbivores (3.3‰ higher). There is no traceof marine protein consumption in their diet, but theisotopic evidence presented here does not exclude theoccasional consumption of freshwater resources. How-ever, the δ15N values of human individuals reflect nosignificant consumption of these resources and high-light the necessity to increase the sample size inregional freshwater fauna to provide more informationon this subject.

Discussion

The values of the Montelirio human group fromValencina–Castilleja are indicative of a homogeneousterrestrial C3 diet. There is no clear isotopic evidence

for C4 plant intake on the diet of this community.The contribution of domesticated C3 plants in the dietcould come from wheat and barley, the most com-monly used cereals during the Neolithic in the IberianPeninsula until the Middle Bronze Age where the earli-est millet (C4 pathway plant) was found as a crop(Rovira, 2007).The animal protein component of the diet consisted

mainly of meat, milk and dairy products from livestock.But some studies (e.g. Hedges and Reynard, 2007;O’Connell et al., 2012; Fraser et al., 2013) suggest thatthe interpretation of a very high animal protein intakein prehistoric farmers could have often beenoverestimated because of some limitations in the dis-tinction of animal-rich diets from plant-rich diets usingδ15N values.To resolve these limitations, we construct a model

with four scenarios (based on the study of Fraser et al.,2013) estimating the animal protein fraction (percent-age) of total dietary protein in the Valencina–Castilleja

Figure 3. Four modelled scenarios estimating the animal protein fraction (percentage) of total dietary protein in the Montelirio human diet. Themodels incorporate an ‘inferred’ plant δ15N value (A), the published cereal and pulse δ15N values from Koufovouno, Greece (Boogard et al., 2013)(B–D) and the Montelirio human bone collagen mean δ15N value (+9.1‰).



human diet (Figure 3). And for it, we also consider thevalue of +4‰ (the midpoint of the usually δ15N offsets+3‰ to +5‰) to represent the ‘trophic level’ differ-ences for the δ15N values and the Δplant–herbivore offsetfor δ15N values.As can be seen in Figure 3, the first scenario (A)

represents the standard model (Hedges and Reynard,2007) in which we have calculated the ‘inferred’plant diet δ15N value by subtracting a fractionation(Δplant–herbivore) amount from the herbivore collagenδ15N value, assuming that humans and herbivores areconsuming plant resources of the same isotopic value.This standard model [Figure 3(A)] predicts that the meanδ15N value ofMontelirio human individuals (9.1‰) indi-cates a diet comprising ≥80% of animal protein. AsHedges and Reynard (2007) point out, in prehistoricagricultural societies, this high proportion of animalprotein is not impossible but requires enough cattle tosustain the community with enough meat and dairyproducts. This high amount of animal protein differsfrom all current agrarian subsistence economies and fitsbetter with known pastoralist economies where milk isa major item of nutrition.The three following dietary modelling scenarios [Fig-

ure 3(B–D)] are constructed using the crops and pulsesδ15N values and applies the same +4‰ enrichment to-wards higher trophic levels. Unfortunately, we couldnot analyse any remain of cereals or pulses fromValencina–Castilleja, and for this reason, we use thepublished δ15N values from the Middle–Late Neolithicsite of Koufovouno, Greece (Bogaard et al., 2013),which could represent the archaeobotanical data forthe Mediterranean environment. Using the Greecearchaeobotanical values, we are able to define the startand endpoint δ15N values for the prediction lines foreach model and, consequently, estimate the animal pro-tein fraction of the human diet. Their starting points arethe consumption of 100% plant protein: the B modelrepresents a human consumption of 100% cereal grain,the C model represents a 100% pulse consumer and theD model represents a 50:50 mixed cereal and pulse con-sumer. The estimated animal protein fractions of humandiet in these scenarios are ≥10%, ≥15% and ≥30%, res-pectively. These three values are not similar to the per-centage of animal protein obtained in the standardmodel, because their starting δ15N values (B: +4.7‰,C: +4.5‰ and D: +3.8‰) are not close to the ‘inferred’plant diet δ15N value (A: +1.8‰). In sum, the animalprotein fractions obtained suggest that these three sub-sequent scenarios estimate human diets with drasticallylower percentages of animal protein and that plants pro-vide most of the protein in the diet. In this sense, thehuman individuals from Montelirio would have a lower

intake of animal protein if a significant consumption ofcereals, pulses or a mixture of both crops had occurred.It is far more plausible that the protein component ofthe Montelirio human diet consisted mainly of thatmixture of cereals and pulses in a regular basis (almost,probably, 70%), as well as animal protein from meat,milk and dairy products (almost, probably, 30%), asthe D model predicted. However, it is important tonote the relevance of regional archaeobotanical datato conduct proper analysis on this subject and the con-sequent human dietary reconstruction. These modelshighlight the importance of the stable isotope analysisof the associated crop materials to not underestimatethe role of cereals and pulses in the human diet.Among the homogeneous human diet observed in

Montelirio, it is interesting to point two cases individu-ally that, despite not showing statistically significant dif-ferences with the rest of the group, present a higherconsumption of animal protein. These individuals identi-fied as 5.Mo6 (UE-112) and 7.Mo11 (UE-107), with10‰ and 9.5‰ δ15N values, respectively, are the twooldest women of the group studied. Our first hypothesissuggests that these two women have, probably, a highersocial status due to their age that could lead to a dietarydifferentiation. This hypothesis could be corroboratedby the lower intake of animal protein (1‰ lower δ15Nvalues) by the younger individuals of the group, allwomen with ages between 18 and 30years. Anotherhypothesis suggests that these lower δ15N values forthe younger women might be more related to their preg-nancies (they are the group most affected by pregnancy)that would produce a little reduction of δ15N values(0.3–1.1‰) from conception to birth (Fuller et al., 2004).Further zooarchaeological, archaeobotanical and

isotopical studies including more regional wild and do-mesticated fauna, and freshwater resources, should helpto clarify the specific proportion of animal protein anddomesticated crops in human diet and the social rolesthat could lead to an increase or a reduction of certainfoods in their diet.

The comparison of Montelirio data with other LateNeolithic–Copper Age series of the Iberian Peninsula

For a better understanding of Copper Age subsistencepatterns in the Iberian Peninsula, we compared theValencina–Castilleja results with other Iberian LateNeolithic–Copper Age series that have been published.The contextualised data and the mean values calculatedfor each site are displayed in Table 4.The Montelirio group was compared with nine

published studies of Late Neolithic–Copper Age sites



Table 4. Stable isotopes results of Late Neolithic and Copper Age human samples from the Iberian Peninsula

Site Country Location IP Period δ13C ± δ15N ± n Comments Source of data

Montelirio,Valencina–Castilleja

Spain South Copper Age �18.3 8.7 This study�19.1 9.0�19.1 8.9�19.9 9.1�19.4 10.0�20.3 8.9�19.5 9.5�19.0 8.9�19.3 8.9�19.3 9.0�20.4 9.2�19.4 0.6 9.1 0.4 11

Avenc dels DosForats

Spain East Copper Age �19.1 10.0 Adults only García et al.,2010�19.1 10.4

�19.1 0.0 10.2 0.3 2Cova La Pastora Spain East Copper Age �19.3 9.7 Adults only McClure, et al.,

2011�19.3 10.6Late Neolithic �19.5 9.0

�19.0 10.0�19.1 9.7�19.6 8.1�19.5 9.5

LN-CA �19.3 0.2 9.5 0.8 7Bolores Portugal West 2880–2620 cal BC 2σ

1963–1745 cal BC 2σ�19.6 9.0 Adult and

adolescents onlyWaterman, 2012

�19.5 9.5�20.8 9.8�19.7 9.6�20.4 8.0�20.4 8.6�20.1 0.5 9.1 0.7 6

TholosBorracheira

Portugal West 3310–2923 cal BC 2σ2199–2035 cal BC 2σ

�19.7 8.9 Adults only Waterman, 2012�19.8 8.6�19.8 0.1 8.8 0.2 2

Lapa Rainha II Portugal West 2852–2497 cal BC 2σ �19.6 8.6 Adults only Waterman, 2012�19.4 8.3�19.7 9.8�19.6 0.2 8.9 0.8 3

Zambujal Portugal West 3333–2936 cal BC 2σ2198–1776 cal BC 2σ

�20.7 8.4 Adults only Waterman, 2012�20.1 10.1�20.4 0.4 9.3 1.2 2

Feteira II Portugal West 3694–3367 cal BC 2σ3264–2894 cal BC 2σ

�20.2 8.4 Adults only Waterman, 2012�19.9 8.6�20.6 7.5�20.4 8.6�20.1 8.2�19.7 9.1�20.7 7.8�20.5 7.6�20.1 8.3�20.3 8.2�20.9 7.6�20.1 8.6�20.0 9.0�20.3 0.3 8.3 0.5 13

Paimogo I Portugal West 3264–2576 cal BC 2σ2901–2488 cal BC 2σ

�21.3 8.1 Adult andadolescents only

Waterman, 2012�21.0 10.3�20.4 9.3�20.8 8.5�19.8 8.8�20.1 8.5�20.0 8.0�20.3 8.1�19.7 7.8

(Continues)



on the Iberian Peninsula: Avenc dels Dos Forats (Garcíaet al., 2010), Cova La Pastora (McClure et al., 2011),Bolores, Cabeço da Arruda I, Cova da Moura,Feteira II, Lapa Rainha II, Paimogo I, Tholos Borracheiraand Zambujal (Waterman, 2012; Waterman et al.,2014). The palaeodiet results based on the mean valuesfor each population are displayed in Figure 4.As revealed in Figure 4, the compared Late

Neolithic–Copper Age groups had a similar diet. The lit-tle dietary groupings observed highlight little differencesbetween areas (west, south and east) that seem more re-lated to their regional environment and their probably

different social/economic factors. The little isotopic dif-ferences between groups are a variation of approximately1‰ for mean δ13C values and 1‰ for mean δ15N values,with the exception of Avenc dels Dos Forats, which has ahigher δ15N mean value, 10.2‰.All the sites compared present a completely C3 ter-

restrial diet that shows no significant isotopic evidencefor aquatic resource consumption; although the west-ern Iberian Peninsula sites were located in estuariesand near the coast, the eastern site Avenc dels DosForats was located relatively close to the coast, andthe southern site Montelirio was located relativelyclose to the Guadalquivir River and its tributaries. Thisinsignificant contribution of aquatic resources observedin the human diet of the communities located near thecoast or estuaries, or near freshwater regions, demon-strates the minor use of these resources in the Neolithicand Copper Age periods in the Iberian Peninsula.According to the study of Waterman (2012) and

Waterman et al. (2014), bone apatite δ13C values(δ13Cap) of human individuals from the Portuguesesites (Figure 1) are more variable than bone collagenδ13C values (δ13Cco), suggesting a dietary differentia-tion in plant consumption. In this sense, several indi-viduals from Cova da Moura, Feteira II and Zambujalhave some C4 sources in their diet, which are visiblein the δ13Cap but are not reflected in the δ13Cco values.This highlights the importance of conducting stableisotope studies from bone apatite, correctly validated(e.g. Ambrose et al., 2003; Lee-Thorp, 2008), to

Figure 4. Plot of δ13C and δ15N average values, with error bars, fromLate Neolithic and Copper Age human remains from the IberianPeninsula.

Table 4. (Continued)

Site Country Location IP Period δ13C ± δ15N ± n Comments Source of data

�20.0 7.9�19.7 10.1�20.3 0.5 8.7 0.9 11

Cova da Moura Portugal West 3635–3372 cal BC 2σ2620–2212 cal BC 2σ


Waterman, 2012�19.3 9.7�19.5 9.5�19.5 9.9�19.5 9.3�20.0 9.1�19.0 10.0�20.6 11.2�20.5 8.7�21.0 9.3�19.9 0.7 9.4 0.9 10

Cabeço daArruda I

Portugal West 3331–2885 cal BC 2σ3005–2634 cal BC 2σ


Waterman, 2012�19.8 8.6�19.5 8.4�19.6 8.8�19.8 9.7�20.3 8.6�19.8 7.8�20.1 8.5�19.8 0.3 8.6 0.5 8

LN-CA, Late Neolithic–Copper Age.



employ linear dietary models such as the one proposedby Kellner and Schoeninger (2007).Overall, the similarities in δ13C values could demon-

strate that these populations practised a similar type ofagriculture and animal husbandry, where the C3 cropsselected for the cultivation grew successfully in the en-vironment of each region and the cattle grazed on aland with similar but not identical vegetation. Theseenvironmental differences between regions could havedirectly affected the diet of the fauna and, conse-quently, could also lead to these slight dietary differ-ences between human communities.It is important to note that Zambujal, Cova da

Moura and Paimogo I have a higher deviation in theirmean values. The intra-population heterogeneity ofthe diet of these societies, specially the variation inδ15N values among individuals, could be explained bythe existence of social roles that could have favoureda sector of the population with a differential protein in-take. However, it is important to highlight the smallsample size of Zambujal that could produce thesedifferences.Finally, the similarities in δ15N values, in conjunc-

tion with δ13C values, indicate that these humangroups present a significant consumption of animal pro-tein from meat, milk and dairy products from terrestrialC3 mammals, as well as C3 plant protein from cerealsand pulses. These animal resources came from thepopulation’s livestock, which possibly ate C3 plantsfrom pastures and meadows. The C3 plant protein came,probably, from a characteristic agriculture composed ofwheat, barley and some beans and possible C3 plantsources gathering in their surrounding areas.

Conclusions

The palaeodietary reconstruction of the human indi-viduals from the high-status burial from Montelirio(Valencina–Castilleja) demonstrates that this commu-nity has a homogeneous diet based on C3 terrestrial re-sources. The principal intake came from C3 animalsources, but the amount of animal protein may be re-duced to ≥30% or maybe less (≥15% or ≥10%) by asignificant consumption of C3 crops, highlighting thatmost of the protein come from C3 plant sources.The comparisons made with published data about

Late Neolithic–Copper Age societies on the IberianPeninsula indicate that the diet of these different com-munities was fairly homogeneous in these periods. Theprotein component of the diet consisted mainly ofmeat, milk and dairy products from livestock as wellas the C3 vegetable component from the characteristic

agriculture of the societies of these periods. This studyalso demonstrates the minor use of aquatic resourcesfrom the Neolithic to Copper Age periods in theIberian Peninsula.The present study is one of the first analyses of the

dietary content of Late Neolithic–Copper Age commu-nities in southern Spain, and more stable isotope datafrom different contexts such as isotopic data fromlower-status burials, including more faunal remains,are needed to clarify those issues commented earlierand to assert these assumptions about the LateNeolithic and Copper Age communities.

Acknowledgements

We would like to thank the directors of the archaeo-logical excavation of Montelirio, Álvaro FernándezFlores and Vicente Aycart, for entrusting us with thepresent analysis. Thanks also to J.C. Espín, J.M. Guijo,R. Lacalle, P. López and A. Pajuelo for providing thesamples. Finally, we would like to thank the two anon-ymous reviewers for their helpful contributions.This work has been supported by ‘Dirección General

de Bienes Culturales de la Consejería de Cultura de laJunta de Andalucía’, ‘Ayuntamiento de Valencina de laConcepción (Sevilla)’ and Generalitat de Catalunya(SGR2009-566).

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Documents

A paleodietary Study of Stable Isotope Analysis from a High-status Burial in the Copper Age: The Montelirio Megaithic Structure at Valencina de la Cncepción-Castilleja de Guzmán,