Quaternary International xxx (2017) 1e11

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Quaternary International
journal homepage: www.elsevier.com/locate/quaint

New evidence for subsistence strategies of late pre-colonial societies

of the mouth of the Amazon based on carbon and nitrogen isotopic
data
Tiago Hermenegildo a, *, Tamsin C. O'Connell a, b, Vera L.C. Guapindaia c,
Eduardo G. Neves d
a
Department of Archaeology and Anthropology, University of Cambridge, Downing Street, Cambridge CB2 3ER, UK
b
McDonald Institute for Archaeological Research, University of Cambridge, Downing Street, Cambridge CB2 3ER, UK
c
Museu Paraense Emilio Goeldi, Bel

em, Brazil
d
rio de Arqueologia dos Tro
Laborato
picos, Museu de Arqueologia e Etnologia da Univesidade de Sa~o Paulo, Sa
~o Paulo, Brazil

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

Article history: The nature of subsistence strategies employed by the past inhabitants of Amazonia has been a widely
Received 10 August 2016 debated topic, however little evidence has been found so far in order to support some of the proposed
Received in revised form hypotheses. This article contributes to this debate by presenting new d13C and d15N data from the human
22 February 2017
populations that occupied the Maraca
region of the mouth of the Amazon river, around 500 BP (years
Accepted 4 March 2017
before present). It directly compares these newly generated results to previously published human
Available online xxx
isotope data from neighbouring Marajo
Island (Marajoara phase, 1600 to 700 BP), as well as other areas in
the lowland Neotropics, in an attempt to build a bigger picture of the dietary habits of the Lower Amazon
Keywords:
Archaeology
pre-colonial populations. The overall results suggest that the populations that occupied the mouth of the
Collagen Amazon after 2000 BP had diets based on the exploitation of sh and a wide range of C3 plant resources,
Stable isotopes as well as possibly having a minor C4 or CAM component. The data presented are also consistent with an
Paleodietary studies emerging consensus that there was no single adaptive pattern for ancient Amazonian populations and
Marac
a tradition proposes that diversied economic strategies based on wild and cultivated plants combined with the
Maize exploitation of faunal resources could have developed over time and sustained long-term successful
patterns of human occupations.
2017 Elsevier Ltd and INQUA. All rights reserved.

1. Introduction determinism, and the notion that domesticated resources and

Subsistence strategies have been a fundamental topic of dis-
cussion amongst archaeologists aiming to understand the occu-
pation patterns, mobility and size of different human populations.
In tropical archaeology, the Amazon has been pivotal in the shaping
of theories with a strong focus on resource acquisition (e.g. cultural
ecology, environmental determinism and historical ecology to
mention a few), mainly because it is one of few places where
populations keep traditional methods of food production.
Early studies and theories proposed for the occupation of the
Amazon were strongly inuenced by the concept of environmental
* Corresponding author.
E-mail addresses: thermenegildo@gmail.com (T. Hermenegildo), tco21@cam.ac.
uk (T.C. O'Connell), veragua@museu-goeldi.br (V.L.C. Guapindaia), edgneves@usp.br
(E.G. Neves).
agricultural practices were the only possible way to sustain large,
sedentary and therefore culturally complex human groups
(Meggers, 1954, 1971; Meggers and Evans, 1957; Lathrap, 1968,
1970, 1977; Carneiro, 1957, 1983). These ideas lead some scholars
to work under the catastrophic assumption that tropical forests
were a counterfeit paradise, a green hell where poor soils com-
bined to high temperatures and humidity would severely hinder
agricultural production and therefore limit human development
(Meggers, 1971). The lower Amazon plays a key historical role, as
Betty Meggers, the main proponent of the environmental limitation
theory, used material culture evidence recovered from her exca-
vations in the region (Meggers and Evans, 1957) in conjunction with
ethnographic evidence from other parts of the Amazon as the basis
for her theories (Meggers, 1954, 1971).
In recent years however, scholars have reached a deeper un-
derstanding of traditional subsistence practices, particularly the

http://dx.doi.org/10.1016/j.quaint.2017.03.003
1040-6182/ 2017 Elsevier Ltd and INQUA. All rights reserved.

Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
2 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11
importance of non-domesticated resources (Clement, 1999) and the
potential of their long-term management to sustain resource sta-
bility amongst living Amazonian populations (Posey, 1985, 1998;
Bale
e, 1994, 2013; Politis, 2009; Rival, 1998, 2012). Based on these
ndings, archaeologists working in the Amazon are now more open
to the idea that, in certain contexts, the domestication of environ-
ments e through the extensive management of non-domesticated
food sources e could have been as important as the domestication
of plants and animals to the maintenance of large and sedentary
populations (Erickson, 2008).
In this article, we contribute a different perspective to the
debate on Amazonian subsistence, using quantitative carbon and
nitrogen stable isotope data from archaeological human remains
recovered from the Maraca
region of the mouth of the Amazon
River, dated to around 500 BP. We consider the available hypoth-
eses in the light of our newly generated isotopic data, as well as
previously published carbon and nitrogen isotopic data from Mar-
ajoara phase burials and other well-established sites in the lowland
Neotropics, in an attempt to build a bigger picture of the dietary
habits of lower Amazon populations.
2. The lower Amazon basin
When it reaches its mouth, after travelling for more than
6000 km down from its headwaters in the Andes, the Amazon River
forms a large estuary owing into the Atlantic Ocean close to the
Equator. Water discharge is ca. 200,000 m3s-1, around 20% of the
total amount of aboveground freshwater on the whole planet. Such
a massive water volume clashes constantly with strong tidal vari-
ations from the Atlantic ocean, with daily ranges of more than 8 m

m, more
in some places, and an inuence that can be felt at Santare
than 400 km from the mouth. Such combined forces create highly
dynamic landscapes prone to drastic changes due to strong erosion
and sedimentary processes. The estuary is a vast area, larger than
Switzerland, and it is composed of a mosaic of landforms including
large islands, such as Marajo
(40,000 sq. km), archipelagos, ood-
plains, high ground and hills on the north bank. Vegetation cover
varies accordingly, ranging from tall evergreen forests to periodi-
cally ooded open savannas. Paleoenvironmental data suggests the
conditions we nd nowadays have not changed signicantly in the
past 2000 years, particularly in the very dynamic environment of
Marajo
Island (Lima, 2008; Smith et al., 2011).
The archaeology of the mouth of the Amazon has been studied
Fig. 1. Areas and sites mentioned in this article. Map A: Ucayali basin (a), Coastal Ecuador (b), the Maya heartland (c), Lagoa Santa (d), Vale do Peruau (e), Sa
Catarina Coast (g) and the Lower Amazon in the highlighted square. Map B: Gruta das Caretas (1), Gruta do Pocinho (2), Teso dos Bichos (3), Monte Carmelo (4) and Matinadas (5).
Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
since the end of the nineteenth century (Meggers and Evans, 1957;
Roosevelt, 1991; Schaan, 2004, 2012) and the data show an almost
continuous occupation sequence from the Middle Holocene on-
wards. Starting with coastal shell-mounds bearing Mina phase
ceramics, which are among the earliest in the New World dating to
ca. 5500 BP (Roosevelt, 1995), the archaeological record of middle
and late Holocene is characterized by the sequential occupation of
distinct cultural groups, as inferred from differences in ceramic
styles and settlement patterns.
Among those groups, there were the mound building societies
of the Marajoara phase characterized by the production of beautiful
and richly decorated polychrome ceramic artefacts, which occupied
Marajo
Island (Fig. 1) during the rst and early second millennia AD
(Meggers and Evans, 1957; Roosevelt, 1991). Throughout the Mar-
ajoara phase, articial residential and funerary mounds were
erected on the top of natural levees associated to riverine paleo-
channels (Rossetti et al., 2009).
After the demise of Marajoara around the fourteenth century
AD, the occupation of the mouth of the Amazon was characterized
by a pattern of high cultural diversity, inferred by different
contemporary ceramic styles and settlement patterns as well
(Guapindaia, 2001; Rostain, 2012; Cabral and Saldanha, 2010). One
such local cultural group produced what are known as Maraca

ceramics, characterized by anthropomorphic and zoomorphic
funerary vessels, which were placed standing inside laterite caves
and rockshelters found along oodplain areas on the north bank of
the Amazon (Guapindaia, 2001).
Since the 1950s, research on Marajoara phase sites has focused
on attempts to understand the patterns of political organization of
the societies that occupied them. On the one hand, authors have
suggested that these were stratied chiefdoms that emigrated from
elsewhere in South America and, upon arriving at the mouth of the
Amazon, did not nd ecological conditions suitable for agricultural
practices that could sustain large population densities (Meggers
and Evans, 1957) meaning that they had to resort, instead, to
practices including the management of wild palms (Meggers,
2001). Direct evidence for this is still lacking however. Other au-
thors have proposed the opposite: that the Marajoara phase was a
local development of the mouth of the Amazon and that these
chiefdom-level societies were made possible by highly productive
activities focused on the cultivation of maize (Roosevelt, 1991), or
the management of aquatic fauna (Schaan, 2008). These hypotheses
are still questionable since they are based on scant indirect dietary
~o Paulo coast (f), Santa
 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11
evidence e mostly plant and faunal macro-remains e as well as
minimal and highly variable stable isotope evidence (described in
detail below).
3. Isotope studies in archaeology
Carbon and nitrogen stable isotopes are widely used in palae-
odietary reconstruction, based on the principle that the signal
found in the consumer's tissues relate to that of their food sources
in a predictable manner, therefore providing reliable direct infor-
mation of an individual's dietary habits (Schwarcz and Schoeninger,
1991; Ambrose, 1993; Lee-Thorp, 2008).
Carbon stable isotope studies are centred on primary producers
and their carbon xation processes. In terrestrial environments,
plants are entirely responsible for the xation of carbon e from
atmospheric CO2 into carbohydrates e through photosynthesis.
However, plants have distinct types of photosynthetic metabolisms
that absorb heavy and light carbon stable isotopes (13C and 12C) in
different manners, thus affecting their relative abundance e a
process known as isotope fractionation e which in turn generates
unique isotope ratios for each type of metabolism.
The vast majority of plants use the Calvin-Benson, or C3
pathway, having carbon isotope values (expressed as d13C) ranging
between
30 and
23, while other plants e mostly in the
Poaceae family such as maize, millet and sugar cane e use the
Hatch-Slack, or C4, pathway. These plants display much higher d13C
values than C3 plants, around
16 to
9 (Hatch and Slack,
1966; O'Leary, 1988; Sharp, 2007). A less common third type of
plants e mostly cacti, succulents and bromeliads e use the Cras-
sulacean Acid Metabolism (CAM) pathway showing values that can
vary between C3 and C4 signals depending on environmental con-
ditions (O'Leary, 1988).
C4 species are most common in semiarid tropical and subtrop-
ical environments, in particular savannas and subtropical grass-
lands, whereas forested tropical regions, such as the Amazon, are
abundant in shade-adapted C3 grasses including high numbers of
C3 bamboo species (Sage et al., 1999). Studies show that in the
Amazon about 60% of all grass species are C4 (Medina et al., 1999),
however they are mostly restricted to seasonally ooded environ-
ments. Some oodplain areas are completely dominated by C4
species including Echinochloa polystachya (Piedade et al., 1991) as
well as a number of species in the Panicoideae subfamily (Medina
et al., 1999).
Besides the lower quantity of C4 grasses, closed canopy tropical
forest such as the Amazon are almost completely dominated by C3
arboreous species. In such dense canopied forests, it is very com-
mon to nd a decrease in the d13C values of C3 plants known as the
canopy effect (van der Merwe and Medina, 1991). This phenome-
non is primarily caused by the combination of low light intensity
and high humidity found below the canopy that results in a high
ratio of stomatal conductance to net photosynthesis, directly
interfering with carbon isotope discrimination (Ometto et al.,
2002). A secondary cause is the alteration in d13C values of source
CO2 trapped under the canopy, however that only accounts for 20%
of the observed variation in leaf d13C values (Buchmann et al., 1997).
Reported values for plants inuenced by the canopy effect are
usually around
30 but values as low as
37 have been re-
ported in denser forest areas (Medina and Minchin, 1980).
All of these different d13C values found in various plants are
maintained in a rather predictable manner throughout the food
chain, showing only a minor fractionation of the order of 1 in
each step of the food chain, although with differences arising due to
differential uptake and utilization of different macronutrient frac-
tions of the diet, e.g. the so-called protein routing (DeNiro and
Epstein, 1978; Ambrose, 1993; Gannes et al., 1998; Jim et al.,
Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
3
2004, 2006).
Carbon stable isotopes can also be used to identify the contri-
bution of marine origin resources to a food web. Marine resources
show a wide range of d13C values, but in general, values tend to be
similar to those of C4 plants (Ambrose et al., 1997; Schoeninger and
DeNiro, 1984). Freshwater resources on the other hand often have
lower d13C values than marine ones. In different studies made using
tissue samples of modern sh from two different regions of the
Amazon, d13C values found are very similar to that of the ubiquitous
C3 plants of the Amazon forest. Near Manaus in Brazil, d13C values of
a single species (Colassoma macropomum) are reported to uctuate
between
29 and
25 (Oliveira et al., 2006), while in Guyana
the results of 15 different sh species vary between
38
and
26 (Watson et al., 2013).
Nitrogen isotope analysis demonstrates an increase in the
15 14
N/ N ratio (expressed as d15N) in animals of successive trophic
levels. Therefore, primary producers will have the lowest values
(0e6) and will gradually increase in the order of 3e5 in each
step of the chain (Hedges and Reynard, 2007; O'Connell et al.,
2012).
Environmental conditions can also affect nitrogen isotope ratios.
Two different areas displayed higher than expected d15N values for
mature non-ooded forests (terra rme forests). Near Manaus,
Brazil, average plant values reach 4.5 1.5 (n 14), while forests
in the western amazon state of Rondo ^ nia, Brazil, plants show an
even higher average of 6.7 1.3 (n 13). However, oodplains
(2.5 1.2, n 10) and sandy scrubland (campina vegetation
(-3.1 3.6, n 5)) near Manaus do show substantially lower
values (Martinelli et al., 1999). The authors believe this effect is
present in many lowland tropical forests where the abundance of
nitrogen combined with years of N losses by fractionating pathways
would cause the high foliar d15N values. In environments where N
appears to be limiting (campina) or in young alluvial soils (ood-
plains) the effect is not observed (Martinelli et al., 1999).
Fire can also alter both soil and foliar d15N values in the years
immediately after burning. Different studies show that in varied
contexts, post-burned vegetation has an increase of 2e8 over
unburned ones (Huber et al., 2013; Grogan et al., 2000). This vari-
ation is comparable, if not greater than reported alterations caused
by cattle manure (Szpak, 2014) and can potentially be an interesting
mean of understanding the relevance of slash-and-burn cultivation
to ancient Amazonians.
Marine fauna can also be identied as a dietary component with
the help of nitrogen stable isotopes. Fish usually have elevated d15N
compared to terrestrial mammals and can easily inuence human
values (Richards and Hedges, 1999; Ambrose et al., 1997;
Schoeninger and DeNiro, 1984). Similar to marine sh, freshwater
ones also have higher d15N values, but since they lack the elevated
d13C found in marine fauna, freshwater sh can be signicantly
difcult recognize as part of human diets, particularly as a minor
component (Hedges and Reynard, 2007). However, in contexts
where zooarchaeology studies demonstrated a dominance of
freshwater resources, human d15N values are quite elevated, further
supporting human reliance on freshwater sh (Bonsall et al., 1997).
4. Diets and stable isotopes in the mouth of the Amazon: two
case studies
Tropical forest environments are notoriously known in archae-
ology for their unfavourable conditions when it comes to the
preservation of biological remains. This led to the inevitable un-
derutilization of most studies that involve such samples, including
dietary stable isotopes studies that rely entirely on bone material.
The Amazon is no stranger to this issue having only a few sparse
carbon and nitrogen isotope results published based on
4 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11

archaeological bone material (van Der Merwe et al., 1981;
Roosevelt, 1989, 1991).
Amongst these few carbon and nitrogen isotope results, nine are
from human burials excavated in different sites e Cachoeira, Teso
dos Bichos, Monte Carmelo, Matinadas and unknown sites (Table 1)
e of Marajo
Island, in the lower Amazon estuary (Fig. 1). All of the
studied remains belong to the Marajoara phase, dated to around
1600 to 700 years BP (Roosevelt, 1991:65).
Despite the variable isotope results (Table 1), Roosevelt inter-
preted the data as reection of a mixed diet, including both faunal
and plant protein, but some individuals had a rather positive ni-
trogen isotope value, consistent with relatively high faunal protein
consumption, and others had a much less positive nitrogen isotope
values, indicating lower faunal consumption and substantial con-
sumption of protein from plants (Roosevelt, 1991: 387). The author
also suggested that some individuals had a carbon isotope ratio
consistent with having maize as a staple in their diets (60% levels),
while others had more positive values consistent with 10e30%
maize consumption.
The carbon and nitrogen variability found in the previously
published results is important. Based on the interpretation that
Some people denitely consumed more faunal protein than others,
and some appear to have eaten more maize than others (1991: 387),
two possible reasons were proposed: either a signicant shift in
economy during the Marajoara phase, or differential access to food
within the same population, since the Marajoara sites and ceme-
teries show a complex patterning suggesting the existence of some
level of socioeconomic differentiation. Unfortunately, as Roosevelt
points out, the small sample size and lack of geographical and
chronological context made it difcult for any further interpreta-
tion of the data.
Macro-botanical remains recovered from the Teso dos Bichos
site show a different scenario. Samples are composed mostly of
seed, bark and fruits from savannah woodland and gallery forest
trees (such as Inga sp., Byrsonima crassiora and Spondias lutea) as
well as a substantial quantity of carbonized palm remains, in
particular palms such as aa (Euterpe oleracea) and tucuma ~
(Astrocaryum vulgare). There were also seeds described as large
kernels, about 5-7 mm long, apparently maize, with ne spongy en-
dosperms (Roosevelt, 1991: 377) that are as of yet unconrmed.
Isotopic analyses of these remains might be informative, however,
these are not available for study at present.
The faunal remains for the Tesos dos Bichos site shows a ma-
jority of small freshwater sh remains such as piranha (Serrasalmus
sp.), trara (Hoplias malabaricus) and tamata
or armoured catsh
(Hoplosternun litoralle). Larger sh such as piraruc (Arapaima
gigas) and aruana ~ (Ostoeglossum bicirrhosum) are also present, but
much less frequent. Mammal and avian bones were extremely rare
in the assemblage and were only found in one particular pit in the
whole site (excavation 3); the excavation report does not provide
details of species or genus for the bones found. Despite the prox-
imity to the Atlantic Ocean, marine or estuarine resources do not
seem to have been consumed at all. The predominance of identied
freshwater sh remains was hence interpreted as consistent with
the d15N values found in the human population (Roosevelt,
1991:387).
As for Maraca
, there are not yet direct dates for any of the human
remains excavated from the sites. However, the presence of a glass
bead in one of the urns and a 360 40 BP date from a nearby
occupation site strongly suggests the Maraca
people occupied the
mouth of the Amazon at the time of the arrival of the Europeans
(Guapindaia, 2008). Dietary studies are also completely absent at
Maraca
, which certainly poses a serious issue for the interpretation
of the isotope data.

Table 1
5. Material and methods

and Marajo
Human stable isotope results from Maraca
. Marajo

data from Roosevelt
1991. Stable isotope values can be retrieved from different animal and
Tradition Site Sex d C () d N () C/N ratio
13 15 human tissues, either from organic or mineral material. The most
commonly used organic remains are bone collagen and in rare


Maraca Gruta das Caretas Indeterminate
18.9 11.5 3.3
cases where preservation permits, hair (or fur). Mineral sources are


Maraca Gruta das Caretas Female
19.0 10.9 3.4


Maraca Gruta das Caretas Indeterminate
18.3 11.3 3.5 mostly bone apatite or tooth carbonates, however teeth are


Maraca Gruta das Caretas Male
17.8 11.9 3.3 preferred due to their higher reliability in the face of contamina-


Maraca Gruta das Caretas Male
18.2 11.3 3.3 tion. The different sources analysed also pose individual limitations


Maraca Gruta das Caretas Female
18.8 11.9 3.6 and interpretations to their results: while bone collagen provides


20.6
Maraca Gruta das Caretas Indeterminate 10.6 3.6 both d13C and d15N results, their carbon comes mainly from


Maraca Gruta das Caretas Indeterminate
18.2 11.6 3.3


ingested protein in the diet, whereas carbon in biological apatite
Maraca Gruta das Caretas Male
18.2 11.9 3.3


Maraca Gruta das Caretas Female
18.2 11.7 3.3 reects whole diet but does not contain any nitrogen (Krueger and


Maraca Gruta das Caretas Female?
18.5 12.0 3.5 Sullivan, 1984; Ambrose and Norr, 1993; Tieszen and Fagre, 1993).


Maraca Gruta das Caretas Indeterminate
22.1 10.4 3.4 This study focuses solely on bone collagen due to difculties in


Maraca Gruta das Caretas Male
18.9 11.3 3.2 obtaining tooth material.


Maraca Gruta das Caretas Indeterminate
19.6 11.0 3.6
The human samples analysed were recovered from the urns


Maraca Gruta das Caretas Indeterminate
19.0 11.3 3.6


deposited in different rock shelter burial sites in the highlands of
Maraca Gruta do Pocinho Indeterminate
19.8 11.8 3.4


Maraca Gruta do Pocinho Indeterminate
18.4 11.6 3.4 the Serra do Laranjal (Guapindaia, 2008). All of the sites excavated
Mean
19.0 11.4 3.4 in the region, including the occupation site that provided radio-
1s 1.1 0.5 0.1 carbon dates, are located within a 1 km radius surrounding Igarape

Marajoara unknown Indeterminate
18.1 12.2 n/a do Lago microbasin, around 80 km from the north bank of the
Marajoara Cachoeira Indeterminate
16.3 10.5 n/a
Marajoara Cachoeira Indeterminate
16.6
9.6a n/a
Amazon river. A total of 38 human samples e all presumed to date
Marajoara Teso dos Bichos Indeterminate
14.3 7.6 n/a from around 500 BP e were collected, but only 17 provided
Marajoara unknown Male
16.9 9.9 n/a acceptable results (Table 1). Samples were omitted if there was a
Marajoara unknown Indeterminate
17.8 10.4 n/a lack of collagen or C/N ratios that were not within the acceptable
Marajoara Monte Carmelo Indeterminate
17.9 10.7 n/a
range (DeNiro, 1985; Ambrose, 1990).
Marajoara Matinadas Indeterminate
15.8 9.4 n/a
Mean
16.7 10.1 Bone samples were processed using a collagen extraction
1s 1.4 1.4 method based on Longin (1971), where 500 mge1000 mg of bone
a
Discarded datapoint due to extreme negative d15N value, which falls outside the
material was sandblasted with aluminium oxide and then demin-
remit of all other published archaeological human nitrogen isotopic data of which eralised in an aq. 0.5 M hydrochloric acid (HCl) solution from 3 to 7
we are aware. days, depending on bone density and size. Once demineralised, the

Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11 5

bone material was rinsed 3 times in deionised water and kept in a
pH3 solution at 75  C for 48 h. Once gelatinised, the supernatant
collagenous solution was removed from the test tube using a 9 ml
Ezee-Filter Separator from Elkay Products (60e90 mm porosity) and
freeze dried for 3e5 days. Once dried, the individual collagen
samples were then sub-sampled in triplicates of 0.7e0.9 mg and
analysed at the Godwin Laboratory, University of Cambridge, using
a Costech elemental analyser coupled to a Thermo Finnigan
MAT253 isotope ratio mass spectrometer. Values presented are a
mean of the sub-sampled triplicates. Stable isotope concentrations
are measured as the ratio of the heavier isotope to the lighter
isotope relative to an internationally dened scale, based on the
international standards of VPDB for carbon and oxygen, and AIR for
nitrogen (Hoefs, 1997). Isotopic results are expressed as d values
(d13C, d15N) in units of permil (). Based on replicate analyses of
international (IAEA: caffeine and glutamic acid-USGS-40) and lab-
oratory standards (nylon, alanine and bovine liver standards),
measurement errors for d13C and d15N are less than 0.2 for both
d13C and d15N values. Comparative modern faunal results used to
contextualise our data (taken from Watson et al., 2013, Oliveira
et al., 2006; van der Merwe and Medina, 1991; values in Table 2)
had their d13C values corrected to 1.5 in order to compensate for
the fossil carbon depletion (Marino and McElroy, 1991).
The standard ellipse and Bayesian analysis used was developed
by Jackson et al. (2011) in order to predict size and relationship of
each isotopic dietary niche. In this approach, instead of the
commonly used bi-plot, results are represented in d-space
(Newsome et al., 2007) that delineate the group's isotopic niche in
an ellipse that contains an estimated 40% of the population. This
methodology is a very useful tool for decreasing the uncertainty of
small sample sizes and has proven to be quite effective in groups of
at least 10 samples (Jackson et al., 2011). All the graphics and
analysis were performed in R using the SIAR package (Parnell et al.,
2008).

Table 2
Faunal reference material used in this study. Modern d13C values corrected to 1.5 in order to compensate for the fossil carbon depletion (Marino and McElroy, 1991). Sources:
1- Watson et al. (2013); 2- Oliveira et al. (2006); 3- Roosevelt (1989); 4- Roosevelt (1991); 5- van der Merwe and Medina (1991).

Taxon N d13C () 1s d15N () 1s Type Origin Source

Riverine Fauna
Arapaima sp. 9
20.4 1.2 11.2 0.7 Modern Guiana 1
Crenichla sp. 4
25.9 1.4 12.6 0.8 Modern Guiana 1
Hoplias malabaricus 2
26.6 2.1 12.6 0.4 Modern Guiana 1
Trachycorystes trachycorystes 4
24.6 0.6 11.1 1.4 Modern Guiana 1
Geophagus surinamensis 2
25.5 1.1 11.6 0.3 Modern Guiana 1
Eigenmannia sp. 1
29.6 10.4 Modern Guiana 1
Apteronotus cf. albifrons 1
29.9 8.6 Modern Guiana 1
Sternopygus sp. 1
28.7 10.3 Modern Guiana 1
Amblydoras sp. 1
25.1 10.5 Modern Guiana 1
Platydoras sp. 1
31.2 8.8 Modern Guiana 1
Chilodus sp. 1
28 11.5 Modern Guiana 1
Hypostomus sp. 1
25.8 11.3 Modern Guiana 1
Ancistrus sp. 5
29.2 1.7 9.7 1.6 Modern Guiana 1
Rineloricaria sp. 3
26.9 3.2 Modern Guiana 1
Curimatid 3
36.6 0.2 8.8 0.5 Modern Guiana 1
C. macropomum rising water 31
24.4 0.5 9.8 0.3 Modern Manaus, Brazil 2
C. macropomum high water 32
27.1 0.2 8.9 0.2 Modern Manaus, Brazil 2
C. macropomum falling water 96
27.3 0.2 9.9 0.1 Modern Manaus, Brazil 2
C. macropomum low water 42
26.4 0.2 10.5 0.3 Modern Manaus, Brazil 2
Unidentied sh 1
24.8 11.1 Archaeological Shahuaya, Peru 3
Unidentied sh 1
24.3 8.1 Archaeological Shahuaya, Peru 3
Unidentied sh 1
22.5 4.3 Archaeological Shahuaya, Peru 3
Unidentied sh 1
23.0 11.5 Archaeological
, Brazil
Marajo 4
Unidentied sh 1
22.8 7.9 Archaeological
, Brazil
Marajo 4

Terrestrial fauna
Tapirus terrestris 1
24.0 2.6 Modern Venezuela 5
Pecary tejacu 1
21.3 3.0 Modern Venezuela 5
Dasypus punctata 1
21.5 3.7 Modern Venezuela 5
Dasypus punctata 1
20.7 3.9 Modern Venezuela 5
Tayassu pecari 1
21.3 4.5 Modern Venezuela 5
Alouata saicula 1
21.9 4.6 Modern Venezuela 5
Mazama sp. 1
22.2 7.5 Modern Venezuela 5
Tamandua tetradactyla 1
22.1 7.5 Modern Venezuela 5
Penelope sp. 1
21.9 7.6 Modern Venezuela 5
Mazama sp. 1
22.3 5.9 Archaeological Shahuaya, Peru 3
Mazama sp. 1
22.3 5.3 Archaeological Shahuaya, Peru 3
Mazama sp. 1
22.4 5.9 Archaeological Shahuaya, Peru 3
Unidentied Monkey 1
21.5 6.1 Archaeological Shahuaya, Peru 3
Unidentied Monkey 1
21.1 5.3 Archaeological Shahuaya, Peru 3
Unidentied Monkey 1
21.1 4.6 Archaeological Shahuaya, Peru 3
Unidentied Monkey 1
21.4 4.5 Archaeological Shahuaya, Peru 3
Cuniculus paca 1
22.2 3.9 Archaeological Shahuaya, Peru 3
Cuniculus paca 1
21.8 5.5 Archaeological Shahuaya, Peru 3
Cuniculus paca 1
21.3 6.5 Archaeological Shahuaya, Peru 3
Cuniculus paca 1
17.3 7.1 Archaeological Shahuaya, Peru 3
Tayassu pecari 1
21.4 5.7 Archaeological Shahuaya, Peru 3
Tayassu pecari 1
19.8 6.7 Archaeological Shahuaya, Peru 3
Sylvilagus brasiliensis 1
21.2 5.9 Archaeological Shahuaya, Peru 3
Unidentied Bird 1
22.5 6.5 Archaeological Shahuaya, Peru 3

Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
6 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11

6. Results and discussion are not as signicant as it might rst seem.

Bone collagen results obtained from the 16 human burials from
Maraca
(Table 1) display mean d13C values of
19.0 (1s 1.1) and
mean d15N values of 11.4 (1s 0.5). At a rst glance, these results
suggest a predominately C3 based diet substantially supported by
animal protein. At the neighbouring Marajo
Island, results from the
multiple sites in the Marajoara period (analysed by Roosevelt, 1991)
show a considerable difference for both d13C (
16.7, 1s 1.4) and
d15N (10.1 1s 1.4) values, indicating that at Marajo
populations
apparently consumed more 13C-enriched foods and less overall
animal protein than Marac
a. The discrepancy between the two
regions is further supported by Mann-Whitney U tests that point to
signicant differences for both d13C (U 2.5, Z
3.59, p < 0.001)
and d15N (U 20.5, Z 2.44, p 0.014).
However, bulk collagen analysis of human material alone is not
sufcient to reach reliable conclusions on their dietary patterns of
any given population. The understanding of faunal and botanical
isotope values are fundamental for contextualizing human diets in
their environment, particularly when considering the wide possi-
bility of environmental variations in stable carbon and nitrogen
values. At Marajo
, only two unidentied sh remains provided re-
sults (Roosevelt, 1991) and at Maraca
region none of the local ex-
cavations has ever recovered plant or animal remains from either
burial or occupation sites. This brings an immediate contextual
issue to the material and in order to mitigate it, faunal results from
modern day studies as well as archaeological fauna from the
Ucayali region, in the upper Amazon basin (Roosevelt, 1989), were
used in this study (Table 2). There are obvious problems involved in
comparing sites that are geographically and chronologically so
distant, but these are the only reported results of archaeological
fauna in the whole Amazon basin.
Terrestrial archaeological fauna from the Ucayali region is
mostly composed of herbivores (deer, rabbits and pacas) and om-
nivores (monkeys and peccaries). Overall results show low values
for both d13C and d15N with little deviation, typical of C3 consumers
(mean for all fauna of
21.3, 1s 1.3 and 5.7, 1s 0.8
respectively, n 15). Modern day fauna from the Venezuelan
Amazon (van der Merwe and Medina, 1991) had very similar results
to the Ucayali data (
21.9, 1s 0.8 and 4.7, 1s 2.0, n 9) and
do not show a signicant difference for either d13C (Mann-Whitney,
U 58, Z 0.53, p 0.59) and d15N values (Mann-Whitney,
U 49.5, Z 1.04, p 0.3). This suggests that, at least for the
material analysed, the above-mentioned distance and period issues
The few archaeological sh results for Marajo
(n 2) and
Ucayali (n 3) were considered together, and displayed low d13C
results with little variation (
23.5, 1s 1.0), whereas the ni-
trogen isotopic data were widely variable across the two sites (8.6
1s 2.9). Modern day sh from two different parts of the Amazon
have also been considered: Guiana, containing 39 individuals of
different species (Watson et al., 2013); and the Amazon River near
Manaus, where 201 individuals of Colossoma macropomun
(commonly known as Tambaqui) were sampled by Oliveira et al.
(2006). Like the archaeological sh material, d13C values were low
(
27.3, 1s 3.3) but d15N values were higher (10.1, 1s 2.0).
Bayesian inference adds further insights for the interpretation of
the dietary context of the lower Amazon (Fig. 2). Terrestrial fauna
once again shows very similar results between the modern day and
archaeological groups, here presented in the form of a substantial
overlap between the two ellipses. This further supports the idea
that isotopic niches appear to be very similar for most potentially
consumable terrestrial fauna throughout the Amazon, but certainly
more studies are necessary to conrm this hypothesis.
Aquatic resources on the other hand show no overlap, either in
the actual data or in the Bayesian ellipses, but that is possibly a case
of population isotopic variability or a result of the small sample size
in the archaeological material (n 5), as the Bayesian model used is
much less consistent below 10 samples (Jackson et al., 2011).
As for the human groups, the standard ellipses representing the
Maraca
and Marajo
populations show a minimal overlap, thus
supporting the signicant difference found for both variables in the
Mann-Whitney U tests.
The d15N values of the Marac
a population shows similar values
to modern day sh results, thus suggesting that sh was a much
more substantial part of their diets than terrestrial animals. It also
shows small d13C increase over modern freshwater and terrestrial
faunal groups, suggestive of carbon input from sources other than
these.
Marajo
also displays lower and more variable human d15N
values (mean 10.1, 1s 1.4), than Marac
a (mean 11.4, 1s 0.5),
yet higher d13C values (
16.7, 1s 1.4 and
19.0, 1s 1.1
respectively). The isotopic data strongly suggest that the diets of
Marajo
and Marac
a are different, both in composition and vari-
ability, although it is difcult to identify the dietary composition
more specically based on our isotopic data alone. The results from
Marajo
do not cluster with either the measured sh (modern or
archaeological) or the terrestrial resources, however there was a

Fig. 2. Scatterplot of d13C and d15N results for faunal and human populations of the Lower Amazon. Standard ellipse area of isotopic niches represent an estimated 40% of the

data from Roosevelt (1991); Ucayali data from Roosevelt (1989); modern sh data from Watson et al. (2013) and Oliveira et al. (2006);
population (after Jackson et al., 2011). Marajo
modern terrestrial fauna data from van der Merwe and Medina (1991). Modern d13C values corrected to 1.5 according to Marino and McElroy (1991). Graphic representation made
in R using the SIAR package (Parnell et al., 2008).

Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11

.
predominance of sh remains found at Teso dos Bichos, Marajo
7. The meaning of the isotopic data from the mouth of the
Amazon
We compared the human isotopic results from the mouth of the
Amazon with isotopic data of humans from other areas in the
Neotropics. Unfortunately, the Amazon has just one other region
with a few carbon and nitrogen isotope results, the Ucayali basin in
Peru (Roosevelt, 1989), so to broaden our dataset we also included
data from sites with well-understood diets from outside Amazonia.
The Ucayali basin assemblage is made of nine samples from
three sites occupied in two distinct periods, 1900 to 1000 BP and
1000 to 500 BP. As Roosevelt (1989) proposes, there seems to be a
dietary shift between these two periods, with the humans from the
earlier period having lower d13C values (
15 to
13) than those
from the later (
12 to
10), thus suggesting a greater reliance on
maize in the later period. The sample size is quite small (n 4 for
early period and n 5 for later period) making it is difcult to
substantiate a dietary shift theory. Because of sample size, and
because the absolute isotopic values of these samples are quite
different from the other Amazonian samples, we combined all the
Ucayali basin samples in a single group for the purposes of the
Bayesian inference model.
Sites along the Ecuadorian coast were also included for having
potential similarities in terrestrial fauna e deer (Mazama sp.) and
peccary (Tayassu pecari and Pecari tajacu) e and cultivated re-
sources e maize (Zea mays), squashes (Curcubita sp.) e to those in
the lower Amazon region (Piperno and Pearsall, 1998). The Ecua-
dorian sites also show a long occupation sequence that allowed
isotope studies to demonstrate a clear dietary shift, as maize
became a staple in the region (van Der Merwe et al., 1993). In order
to better portray the different aspects of the dietary transition, the
Ecuadorian isotope data was divided in three main groups: the
early formative pre-maize group (n 7) containing samples from
the Valdivia 1 and 2 phases (6100e5000 BP); the late formative
group (n 27) comprising the Macharilla and Gungala phases
(3700-2300 BP), when maize was introduced; and nally the late
occupation group (n 15) including the Guangala, Bahia and
Manten ~ o phases (2300e500 BP) used as a well-established
example of a maize-dominated diet supplemented with marine
resources.
Another dataset from outside the Amazon is 135 individuals
from three sites from the Mayan heartland classic period e Holmul,
Seibal and Altar de Sacricios (Wright, 1994; Gerry, 1993). These
results were included since the Maya are one of the only known pre
European contact societies in the Americas to thrive in a lowland
tropical forest environment using intensive and extensive maize
cultivation as a staple in their diets. Despite the distance between
the Amazon and the Pete
n forests, both are tropical forests with
similar environmental contexts therefore sharing many potential
resources. This includes terrestrial fauna, such as Cuniculus paca,
Mazama americana, Tayassu pecari and Pecari tajacu, as well as
cultivated plants including maize (Zea mays), squashes (Curcubita
sp.), beans (Phaseolus sp.), chillies (Capsicum sp.) and manioc
(Manihot esculenta) (Demarest, 2004; Sheets et al., 2012; Flannery,
2014).
Results from shell mounds from Southern Brazil were included
as a proxy for diets with different proportions of marine and
terrestrial resources but no maize or other C4 plant sources. Despite
being in a much higher latitude, between the tropical and sub-
tropical zones, these sites are also located in a densely forested
environment (the Atlantic forests) and shared many potential re-
sources with Amazonia, particularly terrestrial fauna. However
potential environmental differences should be considered in terms
Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
7
of possible impact on the isotope data. The dataset used (taken
from Colonese et al., 2014) was divided in three groups: Moraes
(n 15), dated between 6775 and 6499 and 5289e4887 cal BP, is a
uvial shell mound located around 35 km from the present day
coast of Sa ~o Paulo state and shows evidence of a fully terrestrial
diet; Piaaguera (n 13), dated between 5894 and 5326 and
5887e5314 cal BP, is a shell mound located around 12 km from the
present day shoreline in the same Sa ~o Paulo state and shows a
mixed marine/terrestrial diet; and a third group includes two ma-
rine shell mound sites from the Santa Catarina state, Jabuticabeira II
(n 44, dated between 3137 and 2794 to 1860e1524 cal BP) and
Galheta IV (n 7, dated between 1304 and 1140 and 913e739 cal
BP), both having a very typical marine dominated diet.
Lastly, we included data from the central Brazilian cerrado re-
gion, an open woodland savannah environment, to illustrate the
isotopic data from an open environment completely based on C3
plants and terrestrial resources. The dataset contains four sites in
two distinct regions: Lapa do Santo (n 6) and Lapa das Boleiras
(n 2), dated to around 8500 and 7000 BP, in the Lagoa Santa area;
and Lapa do Boquete (n 5) and Abrigo Malhador (n 5), dated
between 7000 and 4500 BP and 1200 to 600 BP, in the Vale do
Peruau area (Strauss et al., 2016; Hermenegildo, 2009).
Graphic representation of the datasets and their inferred
Bayesian ellipses give a better perspective on the diets of the lower
Amazon (Fig. 3; all groups and mean d13C and d15N values are
presented in Table 3). It clearly shows the Marac
a and Marajo

groups to have lower bone collagen d13C values relative to pop-
ulations that undoubtedly had staple maize diets, such as the
Mayan and Late Ecuador groups, therefore conrming that the bulk
of their diets was very likely based on sources with low d13C values
such as C3 plants, riverine or terrestrial fauna.
Of the two lower Amazon sites, Marajo
displays slightly higher
human bone collagen d13C results, but even its most 13C-enriched
individual (d13C of
14.3) does not seem to be approaching the
carbon isotope ratio of staple corn diets (Roosevelt, 1991:387). In
fact, this result is more similar to the mixed marine/terrestrial diets
found at Piaaguera or Early Ucayali individuals (mean d13C
of
14.7, n 4) believed to have a medium-low carbon isotope
consonant with a diet based on starchy crops such as manioc
(Roosevelt, 1989:52).
Human d15N results from Marajo
are some of the lowest of
studied tropical forest groups, on a similar level to the Ucayali and
Maya groups. As mentioned before, the Maya relied heavily on
maize agriculture as a staple and the Ucayali individuals appear to
follow a similar strategy. This leads us to conclude that the Mar-
ajoara also had a stronger dependence on plant resources, but in
this case, according to the botanic remains from Teso dos Bichos, it
is likely that fruits from the aai (Euterpe oleracea) and tucum~ a
(Astrocaryum vulgare) palms were providing the bulk of the nutri-
tion instead of maize. Despite the limited evidence available, some
researchers also support the idea that palms were a very important
foodsource in the Amazon estuary (Meggers, 2001; Neves, 2012;
Schaan, 2010), notably the aa palm, probably the most highly
productive non-domesticated plant in the lower Amazon. We are
not discarding the possible relevance of manioc (Manihot esculenta)
and wild rice (Leesera hexandra), but considering their absolute
absence in the botanical record from Teso dos Bichos and the fact
that both are C3 plants (van der Merwe and Medina, 1991; Medina
et al., 1999) that cannot be distinguished from a predominantly C3
diet using stable isotope analysis, it would be premature to assume
their importance at this point.
Amerindians from Maraca
show very low d13C results, much like
the Moraes data, Central Brazil and particularly the Early Ecuador
Formative group of which it shows an almost complete ellipse
overlap (Fig. 3). At Ecuador, van Der Merwe et al. (1993) interpreted
8 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11

Fig. 3. Scatterplot of d13C and d15N results for human populations of the Lower Amazon, Ucayali basin (from Roosevelt, 1989), Early Formative, Late formative and Late coastal
Ecuador (from van Der Merwe et al., 1993) and Maya heartland classic period (from Wright, 1994; Gerry, 1993), Central Brazil (from Hermenegildo, 2009), Moraes, Piaaguera and
Jabuticabeira II (Jab II)/Galheta IV (Gal IV) (from Colonese et al., 2014). Symbols: (,) Amazon, (B) Ecuador Coastal, () Maya, () Central Brazil, () Southern Brazil. Standard
ellipse area of isotopic niches represent estimated 40% of the population (after Jackson et al., 2011). Graphic representation made in R using the SIAR package (Parnell et al., 2008).

Table 3 impossible to determine the full extent of slash-and-burn use and

Human groups used in the Bayesian analysis. Sources: 1- Roosevelt (1991); 2- subsequent inuence of the canopy effect in the d13C values of the
Roosevelt (1989); 3- van Der Merwe et al. (1993); 4- Wright (1994) and Gerry
Maraca
people.
(1993); 5- Hermenegildo (2009); 6 e Colonese et al. (2014).
Riverine resources could be equally overshadowing the pres-
Group N d13C () 1s d15N () 1s Source ence of sources with high d13C in the human diet, as modern sh
Maraca
17
19,0 1,1 11,4 0,5 this study results used in this study show very low d13C values (Fig. 2). Un-
Marajo
7
16,7 1,4 10,1 1,4 1 fortunately, there are no sh remains found at Maraca
to conrm
Ucayali 9
12,9 1,8 9,3 1,3 2
this thesis, but if they happened to have similarly low d13C values as
Early Formative Ecuador 7
19,1 1,5 11,7 1,8 3
Late Formative Ecuador 27
11,3 2,2 13,8 1,8 3 modern sh, it is very likely the that input of high d13C dietary
Late Ecuador 15
8,5 1,3 14,2 0,9 3 components (such as C4 plants) for the Maraca
s human population
Maya Heartland 135
9,5 1,2 9,1 1,0 4 is greater than it appears, though modulated by complications of
Central Brazil 18
20,8 1,7 6,4 1,8 5 dietary routing.
Jab-II/G-IV 54
11,5 1,5 17,4 1,5 6
The contribution of sources high in d13C to the Maraca
diets is
Piaaguera 13
15,4 1,1 13,7 0,8 6
Moraes 15
20,8 0,4 10,8 0,5 6 most evident when the results are compared to the purely C3
consumers in the Moraes and Central Brazil groups. Considering
the canopy effect could be providing low d13C resources to the
the low d13C results as a predominately C3 diet with a small Maraca
diets in the form of either sh or wild plants, a purely C3
contribution of marine resources that caused a slight increase in the plant/sh diet in that context should have similar or even likely
d13C values. The similar results at Maraca
indeed lead us to a similar lower d13C results than both Moraes and Central Brazil and not the
interpretation however, it is more difcult to understand the full slightly higher results found (Fig. 3). The Central Brazil results are
extent of the d13C increase given the complexity of the d13C values particularly relevant here since the open canopied, woodland
of all potential dietary sources available in the region. savannah vegetation in the region does not show the d13C lowering
The Marac
a sites are located in a predominantly forested, C3 caused by the canopy effect (Miranda et al., 1997) and yet human
dominated region, that in theory is an ideal scenario to nd very bone collagen results are lower than at Maraca
.
low d13C foliar values caused by the canopy effect (as demonstrated
s human d15N
According to the Bayesian analysis (Fig. 3), Maraca
in van der Merwe and Medina, 1991). Therefore, if animals and results were once again very similar to Early Formative Ecuador
humans were feeding on C3 plants with these very low d13C values, group, thus falling in between the tropical groups with the lowest
any other 13C-enriched resources in the diet e such as marine, CAM d15N values e Marajo
, Ucayali and Maya e and the marine con-
and C4 plants e could be rather underestimated when using sumer groups e Late Ecuador, Late Formative Ecuador, Jab II/Gal IV
collagen based isotope studies without a deeper analysis of the and Piaaguera. This reinforces the idea that the people at Maraca

local ora and fauna. Other potential complications include differ- had a mixed diet, with a stronger riverine component than Marajo
,
ential representation of dietary macronutrients with different iso- likely as a consequence of its location not far from the Amazon
topic values (e.g. marine protein vs C3 carbohydrate: Jim et al., oodplain.
2006). In sum, the lower Amazon diets during the Marajoara and
However, there is also evidence that raises doubt over the direct Maraca
periods seem to be mostly based on C3 plants and riverine
contribution of the canopy effect in the human population of fauna, very likely sh. Both areas show a small input of dietary
Amazonia. van der Merwe and Medina (1991:255, Table 2) found resources with higher d13C values, but with the available informa-
considerable difference between the d13C values of forest tion, it is impossible to determine if maize, marine resources or
(about
35.0) and swidden plants (
24.3 for manioc root) even, however improbable, CAM plants (possibly pineapples, Ana-
from the same area. Therefore, the higher the reliance on slash- nas comosus) were responsible for the observed results. Neverthe-
and-burn techniques a population has, the lesser the direct less, whatever the cause, the human d13C values are such that all
impact of a closed canopy environment on the d13C values of this possible candidates are minor secondary dietary resources, even
given population. Once again, with the information available, it is when considering possible distortion in the observed results

Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
 T. Hermenegildo et al. / Quaternary International xxx (2017) 1e11
caused by extremely low d13C sources; therefore directly opposing
the hypothesis that maize was likely an important resource during
the Marajoara phase (Roosevelt, 1991).
Throughout this study, the limitations of stable isotope analysis
from bone collagen material are quite evident, particularly when
discerning between potential food sources with similar signals
(Layman et al., 2012), a common scenario in the overwhelmingly C3
environment of the Amazon forest. However, tooth enamel based
isotope analysis have the potential of overcoming this issue since
studies relying on d13C and d18O from enamel performed in tropical
Asia have successfully demonstrated the impact of canopy alter-
ations in the isotope ratios of both faunal and human remains
(Krigbaum, 2005; Roberts et al., 2015).
Compound specic isotope analysis of the amino acids con-
tained in collagen is also a very interesting method to resolve many
of the limitations related to bulk collagen analysis. Carbon stable
isotopes from individual amino acids have proven to be a much
more accurate tool than bulk collagen in recognizing the specic
maize component of human diets (Fogel and Tuross, 2003; Honch
et al., 2012) and could potentially be used for the same purpose
in the material presented in this paper. Nitrogen isotopes from
collagen amino acids have allowed researchers to have a much
better understanding of the pathways of nitrogen transfer through
food webs as well as the contribution of marine and terrestrial
protein to human diets (Styring et al., 2010). Considering the fact
that Amazonian freshwater sh have comparable levels of bulk
collagen d15N enrichment to marine ones, if this similarity extends
to their amino acid d15N patterns this technique has a lot of po-
tential to be applied in the Amazon.
Added to tooth apatite and individual amino acid methodolo-
gies, extensive studies of multiple dietary proxies based on animal
and plant remains alongside stable isotope studies of these po-
tential food sources are fundamental future steps in the under-
standing of the enormous environmental complexity and
abundance of possible resources and strategies employed by past
Amazonian people.
8. Conclusions
Populations from the Maraca
and Marajoara cultures occupied
nearby locations in the mouth of the Amazon in the centuries prior
to the arrival of Europeans in South America and, for this
geographical proximity, they were likely exposed to the same po-
tential range of resources. However, carbon and nitrogen stable
isotope analysis demonstrate that despite issues in sample size,
there are still signicant differences in terms of their isotopic di-
etary niches. Isotopic data of humans from both areas suggest a C3
based diet with a low proportion of resources with higher carbon
isotopic values, although the Marajoara have notably higher d13C
values than Maraca
. However, when comparing the results found
amongst known maize consuming populations, they indicate, that
even if present, maize was not a staple in the human diets during
the Marajoara and Maraca
occupations. There are also differences in
the d15N values of both groups, with Maraca
apparently having a
slightly higher reliance on riverine resources than Marajo
.
These results are important because of the growing evidence of
mid-Holocene maize cultivation across lowland South America is
places as far from Mesoamerica such as coastal Uruguay (Iriarte
et al., 2004), coastal Guiana and the Antilles (Paga
n-Jime
nez
et al., 2015). Likewise, maize has been consistently evident in the
archaeobotanical record of later pre-colonial occupations in the
Amazon dating to the last 1000 years before European contact
(Bruno, 2010; Bozarth et al., 2009; Dickau et al., 2012; McMichael
et al., 2015; Watling et al., 2015, 2017). The isotopic data pre-
sented here shows that, even in places where there is
Please cite this article in press as: Hermenegildo, T., et al., New evidence for subsistence strategies of late pre-colonial societies of the mouth of
the Amazon based on carbon and nitrogen isotopic data, Quaternary International (2017), http://dx.doi.org/10.1016/j.quaint.2017.03.003
9
archaeobotanical evidence for maize cultivation (Bozarth et al.,
2009), it may never have become a staple such as elsewhere in
the New World. In other words, the mere presence of a plant in an
archaeological assemblage does not warrant the interpretation of
its wide use and cultivation in the past. Once more, multiproxy
approaches are necessary to a fair assessment of past dietary
patterns.
The data presented is also consistent with an emerging
consensus that there was no single adaptive pattern for ancient
Amazonian populations (Neves, 2013; Piperno, 2011). In such pro-
ductive and diversied environments such as the Amazonia,
recognized as an independent centre of plant domestication, it is to
be expected that different and diversied economic strategies
based on the management of wild and cultivated plants (Clement
et al., 2015) combined with the exploitation of faunal resources
(Prestes-Carneiro et al., 2015) have developed over time and sus-
tained long-term successful patterns of human occupations there.
This paper shows that the wider application of isotopic data in
different archaeological contexts of the Amazon will be funda-
mental for the understanding of such sophisticated knowledge
systems.
Acknowledgments
We would like to thank Catherine Kneale, Mike Hall and James
Rolfe for helping with the isotope analysis; the Dorothy Garrod
Laboratory for Isotopic Analysis (University of Cambridge) for all
the support in this research; Dr. Francisca Alves Cardoso and Regina
Farias for the help with collecting, identifying and sorting the
samples at Museu Paraense Emilio Goeldi. TH would like to thank
the Brazilian National Council for Scientic and Technological
Development (CNPq) for funding his PhD research (grant number
200179/2009-8).
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