Review of Neanderthal Dietary Reconstructions

from Collagen Sampling & Isotope Analysis

Reconstruction of human diets from stable isotope ratios has been common since the 1970s but only a
handful have been undertaken as of yet on Neanderthals. The majority of isotopic dietary reconstructions has
focused on Holocene human and animal specimens, to evidence changing diets over time. While there are
numerous advantages to isotopic studies, whether they have contributed much new information to our modelling
of Neanderthal diet, in and of themselves remains a debatable question. Nonetheless, the purpose they have
served to independently evidence, supporting and even vindicating previous dietary models and estimates is
noteworthy – especially for Neanderthals, a species for which all models and conclusions are continually,
constantly challenged and debated with less regard for the sufficiency of data and more towards the fervour to
consider questions “unresolved.” Diet-driven explanations of change, speciation and extinction, are and have been
quite appealing – particularly given the nature of archaeological assemblages and morphological comparisons in
which extremes, such as adaptations to “fall-back” foods, are over-represented; isotopic studies may, once refined,
resolve these issues. Results from Neanderthals have demonstrated high values consistent with a high trophic
position, often higher than other predators, with subsistence likely dominated by consumption of large herbivores.
Of these studies, many are routinely disregarded by later reviews for various reasons including inadequate
evidence of the contemporaneity with “associated” faunal remains, often highlighted by revised date estimates
from reassessment of site stratigraphy, and methodological/technological advancements revealing early
discrepancies.
The “pioneering” studies of isotopic dietary signatures involved prehistoric woodland Americans over 30
years ago. Since, more detailed studies of the biological pathways of carbon in photosynthesis, controlled diet
experiments, and observations of free-ranging animals have lead to a better, although not yet complete,
understanding. (Lee-Thorp 2008, 925) Some early studies involved strontium/calcium ratios suffered from the
great variability of these elements in plants and overlap between values of browsers and and carnivores
(Sponheimer 2009, 232) Contemporary dietary isotopic analyses are based on the observations that δ13C values of
an animal are very close to that of its diet, δ15N values relate to environment, and stepwise enrichment of these
values through trophic levels. Although oxygen (δ 18O) and sulphur (δ34S) ratios have similar implications, the
majority of studies focus exclusively on nitrogen and carbon. Sulphur, entering animal tissue from mineral fixation
in plants, varies geographically with the underlying geology. δ34S values have been used to detect marine diets and
fix locations. Isotopes of hydrogen and oxygen are distributed due to their behaviour in water and their ratios in
animal tissue indicative of drinking habits and thermo-physiology, likely varying with trophic level. Fossil studies,
including those of Neanderthal fossils, have focused entirely on carbon and nitrogen. (Lee-Thorp 2008, 928)
 Bones and teeth, the best preserved tissues and thereby most common in the record, are sampled for
collagen. Bone and mineral enamel, which turn over regularly in an individual's life history, present long term
averages while tooth enamel and dentine represent the conditions during the juvenile period in which they form.
Controlled feeding studies have shown that collagen δ13C values mainly reflect dietary protein while apatite δ13C
reflect that of the whole diet (Jim et al 2004). Obtaining collagen specimens can be quite destructive to specimens,
particularly in the past, which in addition to the limited number with sufficiently well-preserved collagen, are the
significant barriers to more frequent applications, as isotopic analyses have multiple advantages to more
traditional dietary reconstructions based entirely from archaeological evidence. Lee-Thorp's 2008 discussion of the
state and history of isotopic studies notes that isotopic analysis identifies the food actually eaten, rather than that
preferentially preserved, as the main advantage (925). Richards mentions that faunal assemblages, potentially
representing a single event, cannot necessarily be assumptive of regular and frequent, while isotope studies
demonstrate long-term averages (2000, 7663). On the other hand, isotope studies provide information for a single
individuals, whose habits may or may not be common for the group. The main challenges of isotopic dietary
analysis are the questions of how much, how reliable, and how representative the values given by samples actually
are. Despite achievements in controlling for diagenesis and environmental variations over time and location, the
translation of isotope values to quantifiable dietary components is far from precise. Various dietary elements, in
various proportions, can result in the same “end state”. (Lee-Thorp 2008, 926)

Interpretating isotope values

The critical values in these studies are the ratio of carbon isotopes ( 13C to 12C) and nitrogen isotopes ( δ15N
to δ14N) of animal tissues. The δ values discussed refer to the change from the environmental levels at the time.
The differing kinetic and thermodynamic properties of the isotopes make the heavier isotopes more rare as they
accumulate in the most stable component of a system and are slower to react. The photosynthesis pathways, C 4 or
C3, discriminate differently against carbon isotopes: temperate/woodland C3 plants (trees, shrubs, and cereals)
demonstrate more negative δ 13C values between -24% and -36% while and tropical C4 plants (maize, millet, sugar
cane) demonstrate δ 13C value of roughly -12.5%. Nitrogen isotope values represent both biological fixation and
biosphere processes. Atmospheric nitrogen is globally uniform, with a low 15N composition; the 15N composition of
plants and soils is slightly higher than that of the atmosphere and soil 15N has a weakly inverse relationship to
rainfall. Plants range from 1-4% δ 15N with variation due to aridity, environmental leeching and salinity. In the
15
oceans, recycled nitrate supplies nitrogen and δ N values ranger from 5-6%. Trophic enrichment generally
increases with a stepwise shift of 2-6% δ 15N and the highest δ 15N values tend to result from long marine food-
chains. (Lee-Thorp 2008) Schoeningler (2009) discusses the likely under-appreciation in variation of δ 13C values
resulting from C3 plants, leading to “significant errors when proposing diets in earlier human populations and in
fossil hominins.” (221) Inter-annual and inter-seasonal isotopic effects, causing variation within plant tissues
greater than can be explained by variation in available carbon dioxide, are among the ecological factors meriting
further attention. Further, the rarity of primate studies relative to bovine and cervid is troubling. (222)
Work continues to increase the expected variation of values – within and between ecosystems – and
indicate that “the characteristic spacing between soils, plants, herbivores and carnivores is not constant” (Ambrose
1991, 295) That trophic enrichment occurs is a given; how it does so, and how to distinguish such enrichment from
that of each animals' biochemical processes, needs
further work. Physiological variations between
species, differences in metabolisms and behaviours,
effect tissue values perhaps as significantly as does
diet. Animal tissues are enriched in δ15N relative to
their diet. While mammalian bone and flesh
seemingly demonstrate similar levels, brain, heart,
lungs and plasma have values more negative
(Ambrose 1991, 297). That un-weaned mammals
display higher values than adults is known but the
ages at which weaning occurs in palaeolithic hominin
species has not been conclusively established. This
has presented a problem for comparisons of isotope
values of Neanderthals to anatomically modern
human infants, although the greater problem by far
Figure 1: δ15N vs δ13C values from Marilliac (Fizet 1995, 72)
for all comparative studies is the insufficiently
resolved issue of calibration for geographic and temporal variations in baseline environmental values.

The Studies
The earliest direct analysis of Neanderthal isotope values was published in 1995, by Fizet et al. The team
sampled two Neanderthal specimens from Marillac, France. An adult skull fragment from layer 10 presented a δ 15N

Figure 2: 13C and δ 15N values for all Neanderthal specimens analysed as of 2005 (Bocherens 2005, 77)
Figure 3: (Bocherens 2005, 80)
value of 11.6% and that from layer 9 a δ15N value of 9.3%. Neanderthal values were described as “similar to those
of wolves”. The ecosystem contained wolves and hyaenas, “true” carnivores, as well as omnivorous foxes; foxes'
δ15N values averaged 8.4% while hyaenas and wolves averaged 10.3%. Marine contributions to the Neanderthal
diet were disregarded due to the lack of lithic industry dedicated to fishing, presumed increased distance to the
ocean during the Ice Age, and lack of archaeological remains; further, they believed the δ15N too low to indicate
marine exploitation. While wolves primarily consumed reindeer, Neanderthal diet seems to have been more
diversified, with the likely consumption of horse and bison, in
addition. In initial publication Fizet et al did not attempt to
interpret the difference between the two specimen's values.
In conclusion, they noted that “for Neanderthals, a
generalization to the whole Marillac population of the
conclusions coming from only two measurements is
premature. Indeed, the Neanderthal diet could have varied
according to the prevailing climatic conditions (77)”.
Bocherens et al (1997) sampled various mammal
specimens, including mammoths, from Scladina, Belgium. In
1999, the team published results of Neanderthal sampling
and comparison, intending to test environmental variation of
the Scladina specimen from the Marillac. The resultant δ15N
Figure 4: δ15N vs δ13C values of Les-Rocher-des-
and δ13C values, 10.9% and -19.9% respectively, and were
Villeneuve (black square), other fossil Neandertals
(black squre), Middle Palaeolithic humans (open
circle) from Beauval 2006 (39)
close to those of carnivorous species, suggesting a similar protein source – herbivores from an open environment.
The authors were careful to note the possible contribution of milk to the high δ 15N values of the Neanderthal
sample as the individual sampled was 10 years of age at death. (Bocherens et al 1999) Neanderthal specimens
from Vindija were sampled and published by Richards et al (2000). Samples from Vi-208 and Vi-207 were
submitted it to the Oxford Radiocarbon Accelerator Unit, which in addition to standard AMS dating, prepared the
sample by ultrafiltration, removing degraded collagen and potential contamination. Dates of 29,080 ±400 and
28,020 ±360 BP made them the youngest directly dated Neanderthal specimens from Europe. Unable to extract
collagen from faunal specimens, the supplemented later specimens, carnivores from 23,000 – 26,000 BP from
other (Dolní Věstonice and Milovice) sites and a single herbivore specimen from Brno-Francouszska. Vi-207
presented -19.5% δ13C and 10.1% δ15N; Vi-208, -20.5% δ13C and 10.8% δ15N. Notably low δ15N values of Ursus
spelaeus from Vindija were attributed to either metabolism of hibernation or a high degree of herbivory while the
δ15N values of mammoths from Milovice, higher than other mammals but in line with previous observations of
mammoths, is thought to relate to the mammoth's narrow dietary range as opposed to the breadth of other
herbivores – and may explain the Neanderthals' high δ 15N values. Archaeological evidence does not support
Neanderthals' preferential consumption of mammoths, but it has been suggested that the lack of mammoth bones
in assemblages from habitation floors may be explained by butchering of mammoths taking place off-site and the
meat subsequently transported. The authors concluded, with this study, that scavenging behaviour must have
been secondary to predation in Neanderthal subsistence.
Beauval et al (2006) directly radiocarbon dated and measured the isotope levels of a femur from Les
Rochers-de-Villeneuve Neanderthal, discovered in 2002. The team took a small piece of cortical bone from the
femur which yielded 3.6% collagen with a δ13C value of -19.0%, δ15N of 11.6%, and a C:N ratio of 3.25, all within the
acceptable range. The analysis produced an estimated date of 48,455±1.878 cal BP – older than that previously
received from a hyena bone; as the difference is not considered significant as it is within 2 standard deviations,
the two are still considered roughly contemporaneous. (37) In the same paper they laid out a comparison of these
values to others and concluded that the Neanderthal values
are consistently high, “clustering among the the slightly more
omnivorous of the large carnivores” (40). The Les-Rochers-
des-Villeneuve Neanderthal is among the highest of all
Neanderthals, clustering with two Les Pradelles specimens.
The δ13C values are generally negative, which the authors
attribute to a combination of the habitats of the prey animals
as well as the trophic levels of the individuals being
measured. (40)
The Saint-Césaire Neanderthal was sampled by
Drucker and Bocherens and analysed with the Les Pradelles
specimens in reference to a “multi-site mixing model” by
Figure 5: δ15N vs δ13C values of Les-Rocher-des-
Bocherens et al (2005). The general conclusions, that the Villeneuve (black square), other fossil Neandertals
(black squre), Middle Palaeolithic humans (open
difference between Neanderthal isotopic values and those of
circle), and other mammals (open squares) from
hyeanas was due to the greater consumption of mammoth Beauval 2006 (39)
 and woolly rhinocerous, alongside less
consumption of reindeer, and similar
contributions of bovinae, deer, and horse
for both predators. The high proportions of
these large herbivores in the Neanderthal
diet, relative to that of the scavenging
hyaenas, is taken to evidence a hunting
culture based on “a prey selection [that]
could result from a long lasting dietary
tradition in Europe” (72). Saint-Césaire and
Les Pradelles specimens were directly
Figure 6: (Bocherens 2005, 81)
compared due to their proximity:
similarities in their isotope signatures, in both absolute values of δ13C comparable to wolves and hyaenas,
respectively, and δ15N presenting relatively higher than associated carnivores (79). “Although the chronological
and cultural contexts are different for Les Pradelles and Saint-Césaire Neanderthals, their trophic position seems to
be similar in both cases” (79). Comparison of the Saint-Césaire to the Spy specimen, with Neanderthal δ 15N values
are higher approximately 2% higher than hyaenas' at both sites, present similar broad patterns (79). The Saint-
Césaire to Scladina specimens, separated both geographically and chronologically with contrasting environmental
habitats (the latter more temperate and forested) nonetheless present similarities that contrast to the differences
of their respective associated herbivores' isotopic signatures. The canopy effect of the more closed environment is
noticeable with the more negative δ13C values of herbivores from Scladina. However, the values seem to indicate
that the Scladina specimen subsisted on prey from open steppe rather than from the forest, implying similar
preference, strategy, and across time and space. The mixing model, intended to determine the relative
compositions of Neanderthal and hyaena diets, was based off models developed to compare situations with two
isotopic pairs and was adapted to deal with more than three possible dietary sources by replacing the constant
fractionation rate with a range of estimated prey to predator collagen fractionation values. (75) Given the large
number of unknowns, the margin of error must be considered quite high in this kind of calculation. Of the twelve
specimens with known isotopic signatures at the time of the study, only five were considered due to “chemical,
physiological, and stratigraphical characteristics” (78). The fact that plant contributions were not taken into
account was justified by the fact that collagen represents the dietary consumption of protein, and plant protein
would be nearly invisible in collagen of an omnivorous animal. Had plants, which have a relative 15N depletion,
been consumed in sufficient quantity that their protein factored, compensatory consumption of enriched 15N foods
would have been necessary to maintain the high δ15N values of the Neanderthal specimens. To posit that more
plant consumption required more mammoth consumption, however, may be premature, in my opinion, without
resolution of many of the issues regarding metabolic processes and preferential-uptake by different tissues.
Nonetheless, consistency does appear remarkably evident.

Discussion
All authors note the need, when interpreting Neanderthal isotope values, to establish tightly associated
faunal assemblages in as large a quantity as possible. Some authors advise against multi-site comparisons, even
intra-species, altogether. Nonetheless, inter-site comparisons are frequently undertaken to substitute for
specimens that could not be sampled, or found. Comparison of a study of living species in the Bialowieza Forest
ecosystem to Upper Palaeolithic sites, with the hope to develop a model of isotopic enrichment, demonstrated the
complexity of possible bias between environmental selection of animals and human selection as well as the
relationship of prey species abundance to predator activity – although the range of enrichment values were
relatively consistent. (Bocherens 2003, 51) While most variation has been assumed attributable to climate and
ecology, so comparisons have advisably been limited to a single (or similar) habitat, even within-habitat herbivore
is questionable. That within-habitat variation “cannot be parsimoniously explained by differences in the nitrogen
isotope composition of their diets” (Ambrose 1991, 303) complicates interpretations, returning focus to the
physiological (water consumption and heat stress) potentials. For δ 15N, variation, amino acid synthesis, urea
excretion, and digestive systems are the most likely unexamined causes (Ambrose 1991, 305).
Given the difficulty of ascribing trophic levels, a classification which is perhaps the most basic and intuitive
of dietary interpretations, to definitive ranges of isotope levels and thereby establishing a model, criticism of other
and more complex interpretations is hardly surprising. Inferences regarding proportions and types of dietary
protein are routinely made by measuring the difference in an individual's δ 15N value from that of expected protein
source. (Hedges 2006, 1240) Such models are greatly undermined by factors including the unknown degree of
variation between individuals from the same environment and culture, with similar diets, and the even more
troubling lack of understanding regarding how and if 15N enrichment varies to accommodate dietary stress and
fluctuations. Suggestions that high protein diets tend to result in larger 15N enrichments have been countered with
the opposite; the metabolic processes still a matter of conjecture. (Hedges 2006, 1241) The “evidence presently
available does not resolve these different explanations. Because the dietary animal protein fraction in human
nutrition is model-dependent a clearly outlined model... must be developed” (Hedges 2006, 1248). The most
glaring of unanswered factors is the matter of how marine and freshwater resource consumption alter isotope
values: that marine diets generally represent with high δ 13C values Is accepted while the extent to which high δ
15
N values also indicate the consumption of marine resources is contested.
Until these issues are resolved, all interpretations – much less full dietary reconstructions – should be
considered preliminary at best. The occasional extravagant claims which result from isotopic analyses are
frustrating, if not entirely misinformed. For the most part, they are not in conflict with the archaeological record;
however, the places in which the two provide discrepant models fall within the “range of error”, so to speak, of
isotopic studies, pertaining to that which cannot, as of yet, be necessarily proven. As the archaeological record
provides specific “snapshots” of events and isotope dietary reconstructions are somewhat generalized averages,
the two will need to be considered both alongside and as supplements to each other. Perhaps, at least for now, the
most appropriate way in which to do this is to use the isotope data to “check” the probability archaeologically-
based models.

Conclusion
Isotopic studies have, perhaps finally and unequivocally, positively answered some of the most contentious
questions of Neandethal behavour: Whether Neanderthals were capable of hunting rather than existing by
scavenging, and whether they were capable of engaging in complex hunting strategies sufficient to capture large
game. Attempts to compare Neanderthal isotope values to those of other species, especially anatomically modern
humans, deserve further consideration in light of methodological and theoretical issues yet unresolved, should
they continue to produce null results of significant dietary difference between the groups, theoretical analysis may
be able to settle the nature of behavioural superiority of modern humans. The foremost issue with isotope studies
is the incredibly small sample sizes employed, in many cases a literal handful of fragmentary specimens sampled
and then compared, and the small number of studies. Regardless of other issues, without a wider breadth of data
interpretations remain speculative and will no doubt be further revised in time.

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