Journal of Archaeological Science: Reports 6 (2016) 768–776

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Journal of Archaeological Science: Reports

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Marine food consumption in coastal northern Chilean (Atacama Desert)

populations during the Formative Period: Implications of isotopic
evidence (C, N, S) for the Neolithic process in south central Andes
Marta Díaz-Zorita Bonilla a,⁎, Dorothée G. Drucker a, Pascale Richardin b, Verónica Silva-Pinto c,d,
Marcela Sepúlveda f,e, Hervé Bocherens a,f
a
Fachbereich Geowissenschaften, AG Biogeologie, Universität Tübingen, Hölderlinstr. 12, D-72074 Tübingen, Germany
b
Centre de Recherche et de Restauration des Musées de France C2RMF, Palais du Louvre, Porte des Lions, 14 quai François Mitterand, 75001 Paris, France
c
Department of Human Evolution, Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, 04103 Leipzig, Germany
d
Área de Antropología, Museo Nacional de Historia Natural, Casilla 787, Santiago, Chile
e
Universidad de Tarapacá, Instituto de Alta Investigación, Laboratorio de Análisis e Investigaciones Arqueométricas, Antofagasta #1520, Arica, Chile
f
Seckenberg Center for Human Evolution and Paleoenvironment HEP, Universität Tübingen, Hölderlinstr. 12, D-72074 Tübingen, Germany

a r t i c l e i n f o a b s t r a c t

Article history: Stable carbon, nitrogen and sulfur isotope analyses combined with radiocarbon dates were carried out on human
Received 28 March 2015 and faunal (camelids) hair remains from three archeological coastal sites in northern Chile (Playa Miller 7, Quiani
Received in revised form 12 January 2016 7 and Camarones 15) to assess subsistence patterns between 4000 and 1500 years ago. Sulfur isotopes were
Accepted 24 January 2016
useful to decipher some cases where carbon and nitrogen did not detect significant consumption of terrestrial
Available online 11 February 2016
resources. These isotopic results confirm that the exploitation of marine resources was the main activity for
Keywords:
the inhabitants of the Formative Period, a time of neolithization process in south central Andes. This contradicts
Formative Period previous assumptions on the speed of the shift from the exploitation of marine to terrestrial resources, suggesting
Stable isotope analysis that this transition did not impact the subsistence strategies of this population as fast as previously thought.
Palaeodiet Therefore the “neolithization” of populations during this period did not lead to a significant change in diet. In
Carbon this paper we present data from fifteen individuals, of which thirteen had a diet mainly based on marine
Nitrogen resources, while two showed a more terrestrial-based diet. These different cases among individuals in relation
Sulfur to different access to resources are discussed. Moreover, the results are compared with those obtained from
Hair
other contemporaneous Chilean sites of the same geographical area. This investigation on mummified hair
samples offers new palaeodiet insights into the prehistoric subsistence strategies in northern Chile during a
key transitional period.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction resources is not as clear as in the pure terrestrial C3 context of Europe.

The transition from a hunter–gatherer subsistence strategy to the
adoption of agriculture represents one of the major adaptive shifts in
human history. This process took place at different times around the
world and was also strongly determined by the local environmental
conditions. Therefore, this transition needs to be analyzed independent-
ly in different areas. For example, the transition from the Mesolithic to
the Neolithic period in Europe, according to the isotopic composition
of carbon and nitrogen, coincided with a rapid shift of marine to terres-
trial foods in coastal contexts (e.g., Lubell et al., 1994; Richards et al.,
2003). However, using the same approach in coastal regions of South
America might be more complicated, since the difference in carbon
and nitrogen isotopic signatures between terrestrial and marine food
⁎ Corresponding author.
E-mail address: marta.diaz-zorita-bonilla@uni-tuebingen.de (M. Díaz-Zorita Bonilla).
In South America, the possible consumption of C4 plants and the effects
of aridity on the nitrogen isotopic composition of terrestrial ecosystems
may lead to marine resources masking the carbon and nitrogen isotopic
signatures of terrestrial resources. In this paper, we investigated this
transitional period on the northern coast of Chile using, in addition to
carbon and nitrogen, sulfur isotopes to refine palaeodiet reconstruction
and to estimate the relative proportions of terrestrial vs. marine
resource consumption.
1.1. Cultural and chronological framework
The definition of the Formative Period in the Andean region is based
on several characteristics similar to the Neolithic process in Europe. In
general, this period is associated with sedentary settlements, the devel-
opment of agriculture, the development of ceramics and other techno-
logical innovations, as well as several changes in burial patterns and

http://dx.doi.org/10.1016/j.jasrep.2016.01.026
2352-409X/© 2016 Elsevier Ltd. All rights reserved.
 M. Díaz-Zorita Bonilla et al. / Journal of Archaeological Science: Reports 6 (2016) 768–776
social organization (Lumbreras, 2006; Lavallée, 2006). In the Atacama
Desert in northern Chile, this period has been mainly identified
archeologically in coastal regions and valleys (Rivera, 1994; Muñoz,
2004; Núñez and Santoro, 2011). Until now, evidence for the Formative
Period in the northernmost Chilean highlands has not been found. This
lack of evidence has been interpreted not only as a possible shift of
distribution from groups inhabiting the coast to the highlands
(Santoro and Chacama, 1984), but also as the maintenance of the
hunting–gathering lifestyle in contrast to other groups that were in
the process of change (Sepúlveda et al., 2013). At lowland sites (coast
and valleys), the Formative is chronologically defined between the
dates of 1700 BC and AD 500 and divided in four clear phases: Faldas
del Morro, El Laucho, Azapa for Early Formative (1700–500 BC) and
Alto Ramírez for the Late Formative (500 BC–AD 500). These periods
of significant social and subsistence change occurred after the
Chinchorro culture, which developed in coastal areas based on hunting,
gathering and fishing (Arriaza et al., 2008). The end of the Formative
coincides with the arrival of Tiwanaku influences from the Titicaca
Highlands plateau, where changes are observed in material culture,
social interaction, and daily life (Chacama, 2004).
In the studied region, the Formative is associated with the first
permanent dwellings such as small villages. In addition, this period
corresponds to the intensification of agriculture, mainly the cultivation
of Zea mays, a C4 plant (Rivera, 1994). However, other C3 plants were
also cultivated such as Cucurbita sp., Lagenaria sp., Manihot suculenta,
Ipomoea batatas, Canna edulis, Phaseolus sp. and Chenopodiaceae
(Núñez and Santoro, 2011). Also, this period went through the
development of technologies such as ceramic, copper, gold and silver
metallurgy, basket-making and textile production (weaving technique).
These new elements reflect social differentiation among individuals at
intra and inter-group levels. These processes of change would have
developed locally from the Archaic period, related to the hunter–gath-
erers, until the Formative (Muñoz, 1989, 2004; Agüero et al., 2006;
Nuñez et al., 2006; Núñez and Santoro, 2011). This differs from previous
models, which assumed that the arrival of foreign groups and the
replacement of populations were responsible for these changes
(Rivera, 1975, 1980), although population variability based on
morphological traits has increased during the Formative (Rothammer
and Dillehay, 2009).
As far as funerary practices are concerned, the Archaic funerary
Chinchorro patterns related to body preparation (Arriaza, 1995;
Arriaza et al., 2008) were entirely replaced by mummy bundles that
had diverse grave goods inside. The new funerary practices involved
the construction of shafts or funerary and ritual tumuli with the
presence of flexed skeletons (Muñoz, 2014).
1.2. Use of stable isotopes in the Formative context
Due to the well-preserved state of the hair of the mummies from the
studied sites (Fresnais et al., 2015), this biological material was used for
the isotopic analyses of carbon, nitrogen and sulfur. This analysis allows
the investigation of palaeodiet in past populations, and in particular hair
reflects the diet of the last stage of the individual's life. In contrast, most
previous isotope studies in this region were carried out on bones and
teeth, reflecting longer time periods of an individual lifetime. Diet can
be investigated based on the correlation between diet composition and
carbon and nitrogen isotope abundances, δ13C and δ15N (e.g., Vogel and
van der Merwe, 1977; van der Merwe and Vogel, 1978; DeNiro and
Epstein, 1978, 1981 Jones et al., 1981; van der Merwe, 1982; Katzenberg
and Krouse, 1989; Ambrose and Norr, 1993). According to O'Connell
et al. (2001), hair keratin values differ from bone collagen from the
same individual, with an enrichment of bone collagen in respect to hair
keratin of 0.86‰ (in δ15N) and 1‰ (in δ13C).
Carbon isotopes can be used to discriminate between C3 and C4 plant
consumption, as well as to distinguish between ecosystems (terrestrial
vs. marine) due to the differences in carbon sources (Calvin and Benson,
769
1948; Randson and Thomas, 1960; Whelan et al., 1970; Schoeninger
and DeNiro, 1984). In addition, nitrogen isotope values can also differ-
entiate between marine and terrestrial consumptions (Schoeninger
et al., 1983; Schoeninger and DeNiro, 1984). Since C4 plants exhibit
higher δ13C values than C3 plants, and aridity tends to increase the
δ15N values in terrestrial plants, it might become difficult to distinguish
terrestrial and marine δ13C and δ15N values in contexts such as the
Chilean coast. In contrast, sulfur isotopes are clearly higher in marine
than in terrestrial food resources (MacAvoy et al., 1998), providing a
way to more accurately identify and distinguish between aquatic and
terrestrial origins (Richards et al., 2001; Craig et al., 2006). In our
study, we combined δ13C, δ15N and δ34S, to distinguish between C4
from C3 consumption and decipher the marine food contributions in
the diet of this population.
The last few decades have seen isotopic analyses commonly applied
to the Archaic and Formative Periods (Tieszen et al., 1992; Tieszen and
Chapman, 1992; Aufderheide et al., 1993, 1994, 2002; Macko et al.,
1999; Petruzzelli et al., 2012; Santana et al., 2012; Torres-Rouf et al.,
2012; Roberts et al., 2013; Silva-Pinto et al., 2014; Pestle et al., 2015).
The combination of δ13C, δ15N and δ34S values has previously been ap-
plied to mummified material from the Chinchorro culture (Macko
et al., 1999). However, isotopic analysis on hair has been done for only
four cases from the Late Formative (Silva-Pinto et al., 2014). Therefore,
our contribution aims to add new data about the palaeodiet of the pop-
ulations in a key period in the development of prehistoric communities
in northern Chile.
1.3. Hair preservation state and reliability of isotopic results
This paper focuses on the potential of stable isotope analysis of mum-
mified hair material to investigate past human diets and adaptations to
the environment. In addition to stable carbon and nitrogen analyses,
this study includes the measurement of sulfur isotopes on hair to improve
the discrimination of marine from terrestrial protein consumption. Hair is
more resistant to contamination and diagenesis than bone collagen in
warm and dry conditions. Indeed, the exceptional preservation of the
hair is reflected by the quality of the keratin (%C, %N, C/N ratio and %S).
Moreover, hair allows us to detect the amount of proteins consumed in
the last 6 months prior to the death of the individual (O'Connell et al.,
2001). In contrast, isotopic data from bone collagen correspond to an av-
erage of proteins consumed over the last 10 years (Ambrose, 1990). In
this case, it is quite important to take this factor into account, since our in-
terest is to know if these Formative coastal groups were adapting to new
ways of life, or whether they were continuing traditional subsistence
strategies. Therefore, the only technique to identify the diet of an individ-
ual in their very last months of life is through the analysis of the hair. Com-
parison between hair and bone material, for example, could also help to
track the possible change throughout the entire life of each individual
(Silva-Pinto et al., 2014).
2. Material & methods
The material consists of hair samples from Playa Miller 7, a site dated
from the Formative Period in Northern Chile. New results from two
Archaic period sites (10,500–1700 BC), Quiani 7 and Camarones 15,
are used for comparison. In addition, these results can be compared to
other isotopic data published from previous and contemporaneous
archeological contexts from northern Chile. All the sites presented
here are located on the Pacific coast of Chile (Fig. 1).
2.1. Sites and samples
2.1.1. Playa Miller 7
The archeological investigations were carried out by Guillermo
Focacci during the 1960s (Focacci, 1974). Originally, this site was
defined as a cemetery composed of underground circular structures
770 M. Díaz-Zorita Bonilla et al. / Journal of Archaeological Science: Reports 6 (2016) 768–776

Fig. 1. Map of the sites.

covered by one or two vegetable fiber mats, similar to what has been
observed in the interior walls. Single burials of adults and subadults
have been documented inside the structures in “flexed and seated” po-
sitions, although most of the bodies were found in fetal positions. Some
individuals were wrapped in thick textiles and some in vegetal fiber
matting. In addition, along with these burials, multiple grave goods
were deposited including: basket-making tools, fishing tools, lithic
tools, ceramics, metal objects and color blocks (Sepúlveda et al., 2011,
2013). In terms of chronology, Playa Miller 7 has been assigned to the
Formative based on a single radiocarbon date of 2480 ± 100 BP
(Rivera, 1977). However, Focacci (1974) observed an area from this
site with agricultural and ceramic burials from the Arica Culture
corresponding to the Late Intermediate Period (approximately AD
1100–1450).
2.1.2. Quiani 7
Quiani belongs to a shell midden area located around 8 km south of
Arica. This site was excavated by Junius Bird and then by Grete Mostny
(Dauelsberg, 1974). At Quiani different sectors have been identified,
belonging to Archaic coastal hunter gatherer groups until the Preincaic
Period around the 12th and 15th centuries.
2.1.3. Camarones 15
This site is a large shell midden located on a plateau at the southern
part of the mouth of the Camarones valley adjacent to the Pacific Ocean.
This site has different occupational sectors that yielded one of the most
important funerary remains from the Archaic Chinchorro culture, but
also from the Formative Period (Rivera et al., 1974; Muñoz et al., 1991).
 M. Díaz-Zorita Bonilla et al. / Journal of Archaeological Science: Reports 6 (2016) 768–776
In total, 15 mummy hair and 5 wool objects made of South American
Camelidae hair (one strand per individual specimen) from Playa Miller
7, Camarones 15 and Quiani 7 were purified for analysis (Fig. 2). Hair
samples were preferably used rather than bone collagen due to their
resistance to contamination and diagenesis (O'Connell and Hedges,
1999; Kempson et al., 2003). Moreover, hair was easily sampled,
whereas bone material or teeth were not usually accessible for sampling
for these mummies.
2.2. Laboratory methodology
Hair and wool samples were cleaned using chloroform/methanol
(1:2) for 1 h to remove all contaminating organic compounds. They
were then rinsed twice with distilled water and left to dry, following
the protocol described by Drucker et al. (2010). Samples were analyzed
for elemental (%C, %N and %S) and isotopic analysis (δ13C, δ15N and δ34S)
at the Department of Geosciences, Tübingen University, using a NC2500
CHN elemental analyzer coupled to a Thermo Quest Delta + XL mass
spectrometer. The δ13C, δ15N and δ34S values are reported according to
the international standards V-PDB, atmospheric air (AIR), and CDT,
respectively. The reproducibility for carbon-, nitrogen- and sulfur-
contents was 5%. The reproducibility for δ13C measurements was
± 0.1‰, for δ15N ± 0.2‰ and for δ34S ± 0.3‰. House internal tests
with standards from camel and elk collagen and Sigma Aldrich Collagen
showed that contents of carbon were reproducible to 0.2%, nitrogen to
0.5% and sulfur to 0.01%. All statistical analyses were performed with
Past software, version 3.10 (Hammer et al., 2001).
2.3. Radiocarbon dating
2.3.1. Human and animal hair sample preparation
The protocol for preparation of keratinized samples (e.g., hair,
furs…) that we have used is based on the selective extraction of keratins
from the cortex (Richardin et al., 2011). This method removes the
cuticle compounds that could contain exogenous carbon from external
contaminants. The total treatment takes place as follows: a cleaning
protocol with water and organic solvents; a soft treatment with diluted
acidic and basic solutions (acid/alkali/acid method), similar to the pro-
tocol used for vegetal samples; extraction of keratin by reduction
using dithiothreitol (or DTT) and precipitation with sodium
deoxycholate (DCO) and trichloroacetic acid (TCA).
The dried samples were then combusted to CO2 at high temperature
(5 h at 850 °C) under high vacuum (at 10−6 Torr). The collected CO2
samples were reduced to graphite at 600 °C using Fe powder as catalyst
prior to AMS dating.
Fig. 2. Mummification from Playa Miller 7 (T305) (© C2RMF, P. Richardin).
771
2.3.2. Radiocarbon measurement, calibration, and correction
All measurements were achieved at the AMS facility of the CEA of
Saclay (Moreau et al., 2013). The radiocarbon activity was calculated
by comparing the measured intensities of the 14C, 13C, and 12C beams
from each sample with those of CO2 standards. The 14C ages (in yrs
BP = years before 1950) were calculated by correcting the isotope
fractionation of δ13C, measured by AMS with the 13C/12C ratio. Calendar
ages were determined using the OxCal v 4.2.3 program (Bronk Ramsey,
1995) and with the latest calibration curve SHCal 13 (Hogg et al., 2013).
Calibrated age ranges correspond to 95.4% probability (2σ) and are
expressed in years cal BC or AD.
Regional radiocarbon reservoir effect variations along the coastal
ocean from northern Chile and southern Peru during the Holocene
have been observed between 5180 and 1160 cal BP. Therefore, the
marine reservoir effect was corrected of 226 ± 98 years (ΔR) according
to Ortlieb et al. (2011) and in Table 1, the calibrated dates were
calculated with this correction.
3. Results and discussion
3.1. Results of δ13C, δ15N and δ34S
The carbon and nitrogen elemental composition of the hair samples
ranged from 43.5 to 48.2‰ and from 12.9 to 14.4‰, respectively
(Table 1), and the atomic C/N ratio ranged from 3.7 to 4.3 (Table 1)
showing a mean of 3.9 for both human and animal hair values. The
sulfur elemental composition ranged from 2.7 to 4.3‰ (Table 1).
The mean δ13C for the human group (n = 15) was − 13.9‰ ± 1.0
(1σ), while the mean δ15N is 23.8‰ ± 3.0 (1σ). There was one outlier
individual from Playa Miller 7 (sample 6bis PML 7_S/N 10) which had
a δ13C value of − 16.7‰ and a δ15N value of 15.7‰. For the faunal re-
mains (n = 5), the mean δ13C was −19.3‰ ± 1.1 (1σ) and the mean
δ15N was 10.0‰ ± 1.3 (1σ) (Table 1).
The mean δ34S for the human group (n = 15) is 29.9‰ ± 2.8 (1σ)
while for the faunal remains (n = 5) the average δ34S is 9.7‰ ± 1.8
(1σ). There were two outliers (sample PLM7_S/N 10 and sample PLM
7_T 81_1) from which values between 11.2 and 15.9‰ were observed.
Interestingly, these two individuals come from Playa Miller 7 (Figs. 3
and 4 and Table 1).
The results of δ13C, δ15N and δ34S values in combination with the
radiocarbon analysis showed that all values decreased from the
beginning of the Formative Period (after 3000 BP) showing a mean
δ13C of − 13.4‰ ± 0.9, δ15N of 24.2‰ ± 0.9 and δ34S of 21.3‰ ± 1.2
towards the Late Formative Period with a mean δ13C of − 14.0‰ ±
1.0, δ15N of 23.8‰ ± 3.5 and δ34S of 19.4‰ ± 3.1). (Fig. 4).
3.2. Interpretation of isotopic values in dietary terms
The results show two different categories of isotopic values that are
related to contrasting dietary source intake. For comparison, we are
using the isotopic values for foods from the Arica region published by
Macko et al. (1999). Considering the offset between hair and bone
collagen discussed before, all of the individuals, except for one child
(sample PML 7_S/N 10), show values between 20.7 and 27.6‰ for
δ15N and between − 12.5 and − 14.5‰ for δ13C, which are consistent
with a diet mainly based on marine resources. The group of Camelidae
samples reflects the typical terrestrial values for herbivores consuming
C3 plants (average of δ13C = − 19.3 and δ15N = 10‰) indicating the
endpoint values for the terrestrial environment. Therefore we have
considered +15‰ δ15N and ≥16‰ δ13C to define the marine environ-
ment according to +5‰ in δ15N and +2‰ in δ13C from the terrestrial
endpoint showed by the Camelidae (Bocherens and Drucker, 2003).
The child individual showed δ15N and δ13C values of 15.7 and −16.7‰
respectively, which are compatible with a mixed diet of C3 plants and
terrestrial proteins with a small input of marine resources (Fig. 3). In
our analysis, we cannot observe differences among individuals based
772 M. Díaz-Zorita Bonilla et al. / Journal of Archaeological Science: Reports 6 (2016) 768–776

Table 1
Results of the isotopic analysis.

Sample Site Type Age Sex %C %N C/N δ13C δ15N %S δ34S 14

C Age Calibrated date range (relative area under probability
(yrs BP) distribution)b

PML 7_12816_5 PM H Infant U 46.8 13.6 4.0 −14.2 20.9 3.93 19.1 2153 ± 22a cal AD 52–336 (1.000)
PML 7_S/N 10 PM H Child U 45.5 13.6 3.9 −16.7 15.7 4.01 11.2 2101 ± 22a cal AD 20–214 (1.000)
PML 7_ T81_1 PM H Adult U 44.8 14.2 3.7 −13.4 20.7 3.68 15.9 1658 ± 22a cal AD 640–884 (1.000)
PML 7_Cr 5_2 PM H Adult (37–43) F 45.5 14.3 3.7 −12.9 25.0 3.94 21.2 2235 ± 22a cal BC 11–6 (0.004) cal AD 2–335 (0.996)
PML 7_T306 PM H Infant U 45.5 14.2 3.7 −12.8 25.2 4.18 19.3 2220 ± 30a cal AD 16–360 (1.000)
PML 7_T 119_3 PM H Infant U 46.9 14.4 3.8 −12.7 24.1 3.85 20.9 3880 ± 30 cal BC 2123–1777 (1.000)
PML 7_T 309_2 PM H Adult (22–28) F 48.2 13.0 4.3 −14.0 27.0 3.20 21.3 2040 ± 22a cal AD 233–565 (1.000)
PML 7_Cr 148_2 PM H Adult (40–46) M 47.9 14.3 3.9 −14.5 27.6 3.64 21.3 2414 ± 22a cal BC 341–281 (0.060) cal BC 259–cal AD 29 (0.940)
PML 7_Cr 3c_5 PM H Adult U 46.9 13.8 4.0 −14.2 21.9 3.96 20.8 2648 ± 22a cal BC 726–685 (0.028) cal BC 666–343 (0.960)
cal BC 281–258 (0.013)
PML 7_T 305_3 PM H Adult M 44.6 13.8 3.8 −14.0 25.8 3.32 21.4 2455 ± 22a cal BC 355–53 (1.000)
PML 7_Ca 4_3 PM H Child (8–12) U 44.7 13.3 3.9 −14.4 26.3 3.59 21.2 2335 ± 22a cal BC 167–cal AD 120 (1.000)
PML 7_T 107_2 PM H Adult (37–43) F 47.9 13.6 4.1 −13.9 25.1 3.80 21.1 2278 ± 22a cal BC 132–cal AD 142 (1.000)
QUI 7_T13_1 Q H Adult (27–33) F 47.3 13.3 4.1 −14.6 23.5 3.76 19.7 3098 ± 22a cal BC 1086–806 (1.000)
CAM 15D _C 19_2 C H Adult U 44.2 12.9 4.0 −13.7 25.6 2.81 22.2 3423 ± 22a cal BC 1496–1185 (1.000)
CAM 15C _C 3_2 C H Adult (34–40) F 43.5 13.1 3.9 −12.5 23.5 2.75 22.5 4391 ± 27a cal BC 2831–2811 (0.012) cal BC 2755–2308 (0.988)
PML 7_T 322_1 PM LL – – 47.2 13.7 4.0 −19.8 11.6 2.81 11.1 2313 ± 22a cal BC 401–349 (0.602) cal BC 309–230 (0.398)
PML 7_Cr 3c_1 PM LL – – 47.4 14.2 3.9 −17.3 8.9 3.48 12.1 2390 ± 22a cal BC 512–373 (1.000)
PML 7_Cr 3c_2 PM LL – – 46.1 14.0 3.9 −20.0 9.6 2.99 8.7 2403 ± 22a cal BC 536–527 (0.014) cal BC 523–383 (0.986)
PML 7_Cr 3c_3 PM LL – – 46.9 14.5 3.8 −20.2 8.7 3.19 8.7 2213 ± 22a cal BC 358–272 (0.491) cal BC 262–193 (0.346)
cal BC 185–146 (0.163)
37 PML 7_T 305_1 PM LL – – 47.2 14.4 3.8 −19.5 11.4 3.39 7.9 2225 ± 30 cal BC 364–193 (0.894) cal BC 185–147 (0.106)

Key to table: PM = Playa Miller; Q = Quiani; C = Camarones; H = human; LL = llama (Lama glama); F = female; M = male; U = undetermined sex.
a
Combined result of 2 separate measures (obtained with the command C Combine of the OxCal 4.2 software).
b
Calibrations were performed on unrounded radiocarbon ages and performed using Calib 7.1 using the mixed southern hemisphere SHCal13 (Hogg et al., 2013) and the Marine13
(Reimer et al., 2013) datasets, input parameters include the regional reservoir correction of 226 ± 98 yrs (Ortlieb et al., 2011).

on sex or the age. The δ13C and δ15N values presented here are similar to
those presented by Silva-Pinto et al. (2014) for hair keratin and bone
collagen for the Azapa Valley (Az70 and Az67) from the Late Formative,
with a terrestrial-based diet in the majority of cases. In our study, most
of the individuals showed a clear pure marine diet, while others showed
terrestrial diet with a small intake of marine resources.
The impact of breastfeeding on the isotopic values of two infants
(younger than 18–24 months, samples PLM 7_12816_5 and PLM
7_T306) should be taken into account. Milk consumption leads to
higher δ15N values of about 3‰ in the infants compared to their mothers
(Fogel et al., 1989; Fuller et al., 2006). Such an increase in δ15N values
could be confused with the results of the consumption of marine
resources. Nonetheless, these values reflect the diet of the mother,
that in one case (PLM 7_12816_5) showed δ15N values around 17‰,
which in combination with the δ13C value of − 14.2‰ indicated a
mixed diet similar to the other subadult case.
The analysis of δ34S improved the distinction between terrestrial-
based and aquatic-based diets. The δ34S values revealed a terrestrial
contribution for two individuals (samples 6bis PML 7_S/N 10 and 7
PML_7_81_1), while if using δ13C and δ15N alone only one individual
would be detected with a terrestrial-based diet. These two individuals
are showing δ13C values of − 16.7‰ and − 13.4‰ and δ15N values of
15.7‰ and 20.7‰. In addition, the δ34S values were 11.2‰ and 15.9‰.
In this case we have considered a minimum of +20‰ for δ34S to detect
marine resources consumption (Nehlich, 2015).
Carbon and nitrogen isotopic analyses failed to detect the terrestrial
protein consumption, possibly due to the mixed consumption of C3 and
C4 plants and arid conditions (high δ15N) that resulted in values
compatible with a marine environment. It seems that sulfur stable
isotopes can better detect consumption of terrestrial resources, and
therefore shows that more individuals than previously thought have
consumed C3 plants and terrestrial proteins. However, a C4 component
in their diet cannot be discarded.
3.3. Dietary implications among the analyzed individuals
Previously published isotopic studies and the results presented here
converge to describe a subsistence strategy predominantly based on
marine resources for the coastal inhabitants from northern Chile in
the Atacama Desert (Aufderheide et al., 1993; Petruzzelli et al., 2012;
Roberts et al., 2013; Pestle et al., 2015). However, according to the
results of sulfur isotopes presented here, two individuals, including a
child, have been identified as consumers of a mixed diet of C3 and C4
plants and terrestrial proteins, with a small input of marine resources.
This is opening a new window into the past, allowing a more detailed

Fig. 3. Results of the isotopic analysis of carbon, nitrogen and sulfur.

 M. Díaz-Zorita Bonilla et al. / Journal of Archaeological Science: Reports 6 (2016) 768–776
Fig. 4. δ15N, δ34S, δ13C and 14C dates (yrs BP).
interpretation of the transition to agriculture and the adoption of new
lifestyles during this period.
The consumption of different resources such as C3 and C4 plants and
terrestrial proteins could be explained by the differential access to food
webs that we tentatively link to the mobility patterns of all Formative
communities. Coming from a different area, where agriculture was the
base of the economy, could be very plausible according to the isotopic
results. This is particularly important for the child and the adult with a
terrestrial signal, as it could be reasonably assumed that the subadult
diet depended exclusively on the parents. This alternative hypothesis
of a recently immigrated individual was already suggested by
Petruzzelli et al. (2012) to explain the shifting to agriculture in a region,
where previously they were relying on marine resources. In terms of
social organization, this could be interpreted either as an individual
coming from a possible different social status with access to comple-
mentary dietary sources or as an individual coming from another social
group with a different subsistence strategy not based on coastal re-
sources but more on animals and terrestrial C3 and C4 plants.
3.4. The subsistence strategy during the Formative Period in northern Chile
Several authors have argued that the Formative Period brought
socio-economical changes (Rivera, 1991; Muñoz, 2004; Lavallée, 2006;
Lumbreras, 2006; Nuñez et al., 2006; Alfonso et al., 2007; Núñez and
Santoro, 2011). In addition, there were also changes in the funerary
patterns, where burials now differed from those classically associated
with the Archaic Chinchorro mortuary practices. Among social and
economic changes, one of the most important with a crucial impact on
the diet was the subsistence shift from hunter–gatherer to agriculture
and pastoralism subsistence. Although agriculture during the Formative
was already implemented (Petruzzelli et al., 2012), still some human
groups from the coast did not change their diet towards terrestrial
sources and were maintaining old dietary traditions based on the
exploitation of marine resources. We have no information yet on the
highland groups since body preservation is very poor in this context.
Only three cases are known: one from Patapatane (Santoro et al.,
773
2001), and two from Puxuma 2 (Santoro and Chacama, 1984), but
further investigation is needed.
In order to assess the exploitation of different environments and
their impact on nutrition, sulfur isotope analysis offers a great potential
for tracking past diets. Unfortunately, sulfur isotope analysis has not
been applied as systematically as carbon and nitrogen isotope analysis
in the studied region, albeit for a few sites: Camarones 17, Chin 1,
Morro 1, Maderas Enco, Camarones 15 and Azapa Valley 140 (Macko
et al., 1999; Aufderheide et al., 1993).
Due to the different time records reflected in stable isotopes applied
to hair keratin and bone collagen (O'Connell et al., 2001), we have only
considered sites where human hair has been analyzed for comparison
with our results, which corresponds in total to six cases (Table 2). To
understand changes through time, the dating of the samples have
been divided into Archaic and Formative according to their chronology.
The results reported in Fig. 5 shows an increase of δ34S values,
corresponding to a shift from terrestrial to marine resources from the
Archaic (starting from δ34S 14.3‰ ± 2.6 at Camarones until δ34S
17.5‰ ± 0.3 at Cam17) to the Formative Period (starting from δ34S
19.6‰ at Quiani until δ34S 22‰ ± 1.8 at Camarones), which culminates
in the high input of marine resources found for some of our studied in-
dividuals. The difference in δ34S among our studied sites and those used
for comparative analysis is about 5‰ and there are two clusters clearly
identified. The average δ34S value for the first cluster is around 15‰ for
the terrestrial mammals and plants consumers, while the second cluster
averages δ34S values at around 21‰ for the pure marine consumers.
Indeed the three cases presented in this paper (Playa Miller 7, Quiani
7 and Camarones 15) are those with the highest values of δ34S and
could therefore reflect pure marine diets except for two individuals
from Playa Miller 7. These outliers have a mixed diet of C3 plants, possi-
bly C4 consumption too, and terrestrial mammals, a situation more sim-
ilar to those of the other sites used for comparative analysis (Table 2).
Except for Azapa Valley 140, all sites are located on the coast, there-
fore the access to marine resources was guaranteed for all of them. For
the case of Azapa Valley 140, a diet based on C3 plants and terrestrial
mammals and maybe less marine sources in diet was expected.
However, not all of the studied individuals seem to have the same
774 M. Díaz-Zorita Bonilla et al. / Journal of Archaeological Science: Reports 6 (2016) 768–776
Table 2
List of sites with δ34S (hair) for comparative analysis from Northern Chile.
Site Chronology
Cam-17 Archaic
Chin-1 Archaic
Morro-1 Archaic
Maderas Enco Archaic
Camarones 15 Archaic
Azapa Valley 140 Archaic
proportion of marine resources in the diet and therefore they may have
practiced different subsistence strategies. As a matter of fact, all but two
of the individuals from Playa Miller 7, and all those from Quiani 7 and
Camarones 15 have a pure marine diet, while the individuals from the
other sites might have used mixed resources with a greater input of ma-
rine proteins, while Azapa Valley 140 clearly relied on C3 plants and ter-
restrial mammals, as did the two individuals from Playa Miller 7. The
group of Camelidae wool analyzed in this study confirms the same
value of δ34S in their diet to those of Azapa Valley, where the terrestrial
ecosystem was the predominant resource for the human population.
In addition, we considered the mean isotopic values for the food web
summarized by Pestle et al. (2015). Although they were measured
mainly on bone collagen, we took into account the offset between hair
and collagen, where there is an enrichment of bone collagen to hair
keratin of 0.86‰ (in δ15N) and 1‰ (in δ13C) (O'Connell et al., 2001).
Their results confirm that the δ15N values from the samples analyzed
in the present study fit the expected values for primarily consumption
of marine resources.
Stable isotope analysis of hair keratin gives reliable results for the
interpretation of the diet on the last months of an individual's life.
However, according to Silva-Pinto et al. (2014) who already analyzed
carbon and nitrogen in different hair segments for several individuals
from the Late Formative (Az-70 and Az67), some variations were
identified through hair growth. Therefore, there is the possibility of a
complementary seasonal diet with alternation of marine and terrestrial
resources. Moreover, stable isotopes of carbon, nitrogen and sulfur are
reflecting the input of the protein fraction and are therefore not
accounting for the carbohydrate fraction.
In terms of interpretation, the variability that has been observed at
Playa Miller 7 might not necessarily be linked to individuals from
diverse origins but rather to people who may have access to broader
resources over the year. Our results also complement recently published
work on dental analysis from several individuals from Playa Miller 7
(Watson and Arriaza, 2014). The authors concluded that these individ-
uals did not change their subsistence strategy, which was mainly based
Fig. 5. Comparative analysis of sulfur isotopes by sites and chronology (1σ).
Reference Location Region
Aufderheide et al. (1993) Coast Norte Grande
Aufderheide et al. (1993) Coast Norte Grande
Macko et al. (1999) Coast Norte Grande
Macko et al. (1999) Coast Norte Grande
Macko et al. (1999) Coast Norte Grande
Macko et al. (1999) Coast Norte Grande
on coastal resources as reflected by dental wear and calculus deposits.
The outlier, which is a child, could indicate that even if this individual
was buried on the coast, he could have been coming from a different
area, possibly the valleys, or had access to different kinds of food from
his mother.
The present stable isotope analyses demonstrate the need for
extending such investigations to other archeological sites with the
same methodology in order to compare sites and make inferences
about subsistence strategies from a diachronic perspective. Therefore,
to understand better the use of the dietary resources in northern Chile
at Atacama Desert, the same type of analysis and tissue should be
used from previous and later periods.
The comparative analysis of our results with new radiocarbon dating
could give a more accurate picture of one of the main characteristics of
the Early Formative, such as the change from hunter–gatherer to
agriculturalist subsistence. In addition, this could help to investigate the
timing and the intensity of this process as well as to discuss the impact
of intra-group interaction. Also we may shed light on the population
growth dynamics of these inhabitants from the Formative Period.
Our results, as well as others still in process, in combination with
bioarcheological analyses could contribute to the better deciphering of
the transition between the Archaic and the Formative Period. It is
quite interesting to observe that while some groups are still following
old traditions of coastal resources exploitation, other groups are
introducing at the same time agricultural and pastoralist practices,
although hunting is not yet discarded. These results are even more
interesting if we consider the extent of the mobility of these populations
and the interaction with other areas, which make the Late Formative
more socially complex than previous periods in terms of social
dynamics in the south and central Andes region.
4. Conclusions
The potential of applying sulfur isotope analysis, in conjunction with
carbon and nitrogen isotopes is promising in the tracking of past diets
and this has been demonstrated in this study. The fact that sulfur stable
isotopes can help identify the consumption of terrestrial resources,
whereas a pure marine-based diet could have been inferred from
nitrogen stable isotopes, is quite interesting for some contexts such as
Northern Chilean Formative communities. Moreover, our results
contribute to the debate of the adoption of agriculture and the
consequent changes related to the sedentarization, and the social and
economic implications towards the development of complex societies.
What has been demonstrated with the combination of the analysis of
hair (reflecting later phases of an individual's life), with the application
of carbon, nitrogen and sulfur (allowing discrimination of marine vs.
terrestrial environments) as well as radiocarbon analyses, is that this
change of subsistence strategy was adopted later than previously
thought, as confirmed by other recently published research. A re-
analysis and reinterpretation of the sites is therefore needed as well as
the development of systematic sampling of carbon, nitrogen and sulfur
isotopic analyses in combination with radiocarbon dating and the
archeological record. This will help to contextualize the remains from
this region and provide a better understanding of human adaptation
to the environment and changing subsistence practices.
 M. Díaz-Zorita Bonilla et al. / Journal of Archaeological Science: Reports 6 (2016) 768–776
Acknowledgments
The authors would like to thank Christoph Wißing and Theda
Himpel for their assistance during the laboratory process and Eva
Alarcón García for the image processing. Thanks also to James Fellows
Yates for proofreading the paper. We want also to thank Bernard Ber-
thier and his team from the Laboratory of Measurement of Carbon 14
(LMC14 — UMS2572) for the graphitization step of the gas samples
and the 14C measurements, and Nathalie Gandolfo and Alice Gimat
from the C2RMF for the sample preparation. Thanks to the Museo
Universidad de Tarapacá — San Miguel de Azapa for giving access and
permission to analyze the archeological material. We would finally
like to thank Proyecto UTA 3711-14 and to the Convenio de Desempeño
Universidad de Tarapacá — MINEDUC.
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