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nature ecology & evolution
Article
https://doi.org/10.1038/s41559-023-02318-z
Stable isotopes show Homo sapiens
dispersed into cold steppes ~45,000 years
ago at Ilsenhöhle in Ranis, Germany
Sarah Pederzani 1,2 , Kate Britton1,3, Manuel Trost1, Helen Fewlass 1,4,
Nicolas Bourgon1,5, Jeremy McCormack 1,6, Klervia Jaouen1,7, Holger Dietl8,
Hans-Jürgen Döhle8, André Kirchner 9, Tobias Lauer1,1 0, Mael Le Corre3,11 ,
Shannon P. McPherron 12, Harald Meller8, Dorothea Mylopotamitaki1,13,
Jörg Orschiedt 8, Hélène Rougier 14,15, Karen Ruebens 1,13 , Tim Schüler 16,
Virginie Sinet-Mathiot 1,17, Geoff M. Smith 1 ,18, Sahra Talamo 1,19,
Thomas Tütken 20, Frido Welker 21, Elena I. Zavala 22,23, Marcel Weiss 1,24 &
Jean-Jacques Hublin1,13
The spread of Homo sapiens into new habitats across Eurasia ~45,000years
ago and the concurrent disappearance of Neanderthals represents a
critical evolutionary turnover in our species’ history. ‘ Tr an si ti onal’ t ec-
hnocomplexes, such as the Lincombian–Ranisian–Jerzmanowician (LRJ),
characterize the European record during this period but their makers
and evolutionary signicance have long remained unclear. New evidence
from Ilsenhöhle in Ranis, Germany, now provides a secure connection of
the LRJ to H. sapiens remains dated to ~45,000years ago, making it one
of the earliest forays of our species to central Europe. Using many stable
isotope records of climate produced from 16 serially sampled equid teeth
spanning ~12,500years of LRJ and Upper Palaeolithic human occupation
at Ranis, we review the ability of early humans to adapt to dierent climate
and habitat conditions. Results show that cold climates prevailed across
LRJ occupations, with a temperature decrease culminating in a pronounced
cold excursion at ~45,000–43,000cal. Directly dated H. sapiens remains
conrm that humans used the site even during this very cold phase.
Together with recent evidence from the Initial Upper Palaeolithic, this
demonstrates that humans operated in severe cold conditions during many
distinct early dispersals into Europe and suggests pronounced adaptability.
The Middle to Upper Palaeolithic transition marks an important period
in human evolutionary history, with the dispersal of Homo sapiens
across Eurasia and the disappearance of other hominins such as Homo
neanderthalensis from the fossil record. Archaeological and genetic
evidence increasingly demonstrates that this transition involved a com-
plex patchwork of archaeological and biological turnovers including
many dispersals of H. sapiens1–5 (but see ref. 6). To interpret the evolu-
tionary significance of these events, it is crucial to determine the envi-
ronments and climatic conditions that H. sapiens groups encountered
during these dispersals.
Prominent models have suggested that early range expansions
of H. sapiens during the Late Pleistocene were linked to warm climatic
Received: 27 June 2023
Accepted: 19 December 2023
Published online: 31 January 2024
Check for updates
A full list of afiliations appears at the end of the paper. e-mail: scpederz@ull.edu.es
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(Supplementary Table 2). The radiocarbon dates from the Hülle faunal
collection show a large spread of ages for the layers labelled ‘brown’
(Supplementary Fig. 1). The LRJ grey layer (X) has more consistent ages,
ranging from 45,900 to 42,100cal, which overlaps with the date
range of the LRJ layer 8 but also with layer 7 of the TLDA/MPI-EVA exca-
vation
13
. Quality control indicators demonstrate exceptional collagen
preservation and purity (Supplementary Table 2) and thus this overlap
should be attributed to the excavation methodology of the 1930s and
the documentation quality of the Hülle faunal collection. To avoid
stratigraphic attribution errors, all analyses in the following are made
only on the basis of the direct radiocarbon dates of the equid specimens.
Stable isotope analyses
Oxygen isotope measurements of sequentially sampled tooth enamel
phosphate (δ
18
O
phos
) show sinusoidal seasonal cycles in all specimens,
with high δ
18
O
phos
peaks representing summers and low δ
18
O
phos
troughs
representing winter inputs (Supplementary Fig. 2, Supplementary Text
2 and Supplementary Table 3). The
87
Sr/
86
Sr values of equids (0.7090–
0.7120), undertaken to confirm that δ18O values are representative of
local conditions without bias from long-distance migrations, fall into
the range of bioavailable values of Thuringian lithological units and
match those observed in hyenas and ursids with typically small to mod-
est home range sizes (Supplementary Text 3, Supplementary Figs. 3 and
4 and Supplementary Table 4). Seasonal
87
Sr/
86
Sr intratooth differences
are mostly very small (<0.0005), with no systematic changes through
time (Supplementary Fig. 3). One individual, R10131, shows a slightly
larger seasonal change (0.0008) and also shows a larger seasonal dif-
ference in δ66Zn (Supplementary Figs. 5 and 6), an isotopic tracer that
reflects some impacts of underlying bedrock type—in addition to the
more prominent dietary effects (Supplementary Text 4). Overlap of
87Sr/86Sr values of this specimen with regionally expected values means
that long-distance movement remains unlikely but cannot be ruled out
entirely. Thus, excluding this specimen from climatic interpretations
is the most cautious approach. Furthermore, the correlation between
87
Sr/
86
Sr and δ
66
Zn seems driven by two outliers and has a very shallow
slope, suggesting that δ
66
Zn values are predominantly driven by diet
(Supplementary Figs. 7and 8, Supplementary Text 3 and Supplemen-
tary Text 4).
Seasonal and mean annual δ18Ophos values show distinct changes
over time (Fig. 1 and Supplementary Table 5), starting with mean annual
values of ~12–13.5‰ at 48,000–45,000cal, then dipping by ~3‰ to
~9–10‰ at ~45–43 kacalBP. After this, δ18Ophos values rise back up to a
similar level at around 42,500cal, followed by a gap in the data and
a phase of high δ
18
O
phos
variability at ~39–36.5 kacal. During the low
δ18Ophos excursion at ~45–43 kacal, winter δ18Ophos values fall as low as
9.0‰, while summers only reach 13.1‰ at their highest value. Seasonal
amplitudes of δ
18
O
phos
(summer–winter differences) range between
0.9‰ and 4.1‰ overall and are highest at ~45–43 kacal with a mean
seasonal amplitude of 3.3±0.8‰. Seasonal δ
18
O
phos
amplitudes are
lower in periods of comparatively higher δ
18
O and correlate negatively
with winter δ
18
O (P=0.0049, R=−0.39, n=50, Pearson correlation)
but not with summer δ18O (Supplementary Fig. 9). Hence, changes in
seasonality are driven predominantly by changes in winter δ
18
O. The
lowest point in δ18Ophos at ~45–43 kacalBP coincides with the highest
dentine and mandible bone collagen δ15N values of 8.7–6.8‰ (Fig. 1) and
the two proxies show a statistically significant correlation, particularly
during this time (Supplementary Figs. 10 and 11, for all data R=−0.36,
P=0.045, n=32; for >42,000cal, R=−0.59, P=0.012, n=17, Pearson
correlation). After the ~45–43 kacalBP high point, δ15N values decline
steadily by ~4‰ until they fluctuate between 3.4‰ and 4.5‰.
Carbon stable isotope values of dentine and mandible bone col-
lagen change little over time, with most individuals falling into a range
of less than 1‰ (−21.3‰ to −20.6‰; Fig. 1). The δ
13
C values do not show
any statistically significant correlations with the other isotope systems
(Supplementary Fig. 10, Pearson correlation). Across a variety of taxa in
phases that facilitated adaptation to higher latitudes7,8. Recent pal-
aeoclimatic data generated directly from archaeological sites have
challenged this idea
9,10
but the scarcity of data from a few sites and
archaeological technocomplexes means that climatic scenarios of
dispersals still lack the complexity that is emerging from the genetic
and archaeological records.
Here we add local palaeoclimatic data pertinent to a newly docu-
mented early incursion of Late Pleistocene H. sapiens into central
Europe. We use multiple stable isotope analysis of faunal remains
from Ilsenhöhle in Ranis (hereafter, Ranis), Germany, to document
the climatic and environmental conditions H. sapiens faced during this
dispersal, associated with the Lincombian–Ranisian–Jerzmanowician
(LRJ) transitional technocomplex11–13. Ranis is located in the Orla valley
(50°39.7563N, 11°33.9139E, Thuringia, Germany; Extended Data
Fig. 1) and is a type site of the LRJ, an archaeological phenomenon of
the Middle to Upper Palaeolithic transition, that extends across north-
ern and central Europe. New genetic, proteomic and chronological
evidence now links the LRJ with directly dated H. sapiens remains at
Ranis and documents one of the earliest dispersals of our species into
the European continent
13
. Using the isotopic data generated here we
demonstrate the climatic and environmental conditions that pioneer-
ing H. sapiens groups exploited during their initial spread across central
and northwestern Europe.
Originally extensively excavated by W. Hülle from 1932 to 193814,
recent re-excavations from 2016 to 2022 (Thuringian State Office
for Preservation of Historical Monuments and Archaeology (TLDA)/
Weimar and the Department of Human Evolution at the Max Planck
Institute for Evolutionary Anthropology, Leipzig (MPI-EVA) excava-
tions) have allowed a state-of-the-art reassessment of the stratigraphy
and chronology of Ranis (Extended Data Fig. 1 and Supplementary
Text 1). In the TLDA/MPI-EVA excavations, the LRJ occupations are
associated with layers 9 and 8, dating to 47,500–45,800cal and
46,800–43,300cal, respectively, with the main occupation in layer
8 (ref. 13). H. sapiens fossil remains were identified in layers 9 (n=1)
and 8 (n=3) and the Hülle collection (n=9), with direct
14
C dates of
the Hülle specimens matching those of layers 9 and 8 (ref. 13). Zooar-
chaeological, archaeological and sediment DNA data suggest that the
LRJ occupations were ephemeral and low-intensity, with most faunal
remains accumulated by carnivores15. The LRJ layers are bracketed by
Upper Palaeolithic deposits above and potentially Middle Palaeolithic
deposits with low artefact density at the base of the sedimentary profile
(Extended Data Fig. 1).
Here, we apply oxygen, carbon, nitrogen, strontium and zinc stable
isotope analyses to directly 14C-dated Equus sp. teeth (enamel bioapa-
tite and dentine and mandible bone collagen) from the transitional LRJ
and Upper Palaeolithic occupations (layers 9–6) to generate evidence
of seasonal palaeotemperatures, water availability and changes in
vegetation cover experienced by the humans that produced the LRJ
record. Fossil teeth were obtained from the Hülle collection (n=14)
and the TLDA/MPI-EVA excavation (n=2; Supplementary Table 1).
Furthermore, 24 tooth specimens representing a variety of herbivore,
omnivore and carnivore taxa were chosen from the Hülle collection for
further δ66Zn and 87Sr/86Sr analyses to explore their feeding ecology
and mobility (Supplementary Table 1). The stratigraphic layers of the
two excavations are clearly correlated and archaeologically equivalent.
However, challenges with the documentation of the Hülle faunal collec-
tion often prevent clear stratigraphic assignments of faunal specimens
(Supplementary Text 1). For this reason, all equid remains studied here
were directly
14
C-dated and correlated to the LRJ and H. sapiens fossils
through the obtained ages.
Results
Chronology
Direct radiocarbon dating of 16 equid specimens yielded calibrated
14
C ages ranging from 48,800 to 36,300cal, covering ~12,500years
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12
13
14
15
10
11
12
13
14
9
10
11
12
13
δ18Ophos (‰ VSMOW)
Mean annual δ18 O Summer δ18 O
Summer Mean annual Winter 1932−1938 2016−2022
3
4
5
6
7
8
9
δ15N (‰ AIR)
−22.0
−21.5
−21.0
−20.5
δ13C (‰ VPDB)
δ13 C δ15 N Winter δ18 O
36,000 38,000 40,000 42,000 44,000 46,000 48,000
0.8
1.0
1.2
1.4
Years cal BP
δ66Zn (‰ JMC Lyon)
Summer/winter δ66 Zn
Excavation
*
*
*
*
*
Fig. 1 | Oxygen, nitrogen, carbon and zinc stable isotope analyses of directly
dated equid teeth show changes in climate and environment through the
LRJ and Upper Palaeolithic sequence of Ranis. Summer peak, mean annual and
winter trough oxygen isotope values show low values throughout the sequence
and a temperature decline from ~48 kacal to a temperature minimum at
~45–43ka cal. This oxygen isotope minimum coincides with high δ15N (dentine
and mandible bone collagen) and δ66Zn values, suggesting a hypergrazer niche
of equids in open steppe environments or very dry soil conditions similarly
indicative of an open environment. This is supported by high δ13C (dentine
and mandible bone collagen) values consistent with a steppe or tundra biome.
One individual has been marked with an asterisk as it has been excluded from
climatic interpretations because 87Sr/86Sr and δ66Zn seasonal amplitudes are
high enough that a seasonal movement cannot be completely excluded. Oxygen
isotope data points represent δ18O summer peak, winter trough and annual
means of individual annual cycles represented in sinusoidal δ18O time series
obtained from sequentially sampled tooth enamel (marked in Supplementary
Fig. 2). Stable isotope data are presented as the mean±measurement uncertainty
based on sample replicates (1s.d., nreplicates=3 for δ18O and nreplicates=2 for all
other proxies where error bars are present; replicate measurements represent
repeated isotopic measurements of aliquots of each single prepared sample).
Measurement uncertainty for δ66Zn is smaller than the symbol size. Horizontal
error bars indicate the 95% calibrated age range of direct radiocarbon dates
(n=1 tooth sample for each data point). Symbol shapes indicate the excavation
origin from either the Hülle (1932–1938, circles) or TLDA/MPI-EVA (2016–2022,
triangles) campaigns. Collagen analysed for δ13C and δ15N was obtained from
tooth dentine for all 1932–1938 samples and from adhering mandible bone for
the two 2016–2022 samples marked by triangle shapes. Stable isotope delta
values are reported in relation to the relevant scale-defining reference materials
Vienna Mean Ocean Water (VSMOW), atmospheric N2 (AIR), Vienna Pee Dee
Belemnite (VPDB), and Johnson Mattey zinc metal (JMC Lyon).
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the food web, δ66Zn values of the Ranis fauna follow expected dietary
and trophic patterns, with the lowest values observed in carnivores and
highest values in herbivores (Supplementary Text 4 and Extended Data
Fig. 2). Within herbivores, woolly rhinoceros (Coelodonta antiquitatis)
show the lowest values, followed by typically browsing or mixed feed-
ing cervid taxa, while equids show the highest δ66Zn values (0.79‰
to 1.51‰). Across time, δ66Zn values of equids are highest at ~45–43
kacal, coinciding with the lowest δ
18
O values and highest δ
15
N val-
ues (excluding R10131 ; Fig. 1), resulting in a positive correlation with
δ
15
N (R=0.43, P=0.0013, n=15, Pearson correlation) and a negative
one with δ
18
O in the time before 42,000cal (R=−0.58, P=0.015,
n=17, Pearson correlation; Supplementary Figs. 11 and 12). However,
diachronic δ66Zn change is relatively small and equid δ66Zn values
never overlap with non-herbivore taxa or even some of the lower-δ66Zn
herbivores such as woolly rhinoceros or cervids (Extended Data Fig. 2).
Water oxygen isotopes and palaeotemperatures
Reconstructed δ18O values of drinking water (δ18Odw), which enable
comparisons with modern meteoric water sources and those of other
fauna, fall systematically below modern day δ18O of local precipitation
(δ18Oprecip), as well as Thuringian rivers and springs (Extended Data
Fig. 3 and Supplementary Table 6). Summer δ
18
O
dw
partially overlap
with modern spring and river water, as these seasonally more buffered
water bodies represent amount-weighted annual averages of δ18Oprecip.
The lowest δ
18
O
dw
values at ~45–43 kacal fall more than 5‰ below
the mean annual δ18O of the modern water sources and more than 8‰
below winter water source values.
This is mirrored by air temperature estimates (Supplementary
Table 6), which fall substantially below modern-day conditions with
the largest difference in winter (Fig. 2). During the ~45–43 ka cal
cold phase, air temperature estimates are lower than modern day
by 7.3±3.6°C in summer, 11.3±3°C for mean annual conditions and
15.3±4.5°C in winter. In the oldest data (~48–45 kacal), temperature
estimates, while less extreme, still fall 3.0–8.3°C below modern-day
temperatures across all seasons. Temperature seasonality ranges from
22.2±2.6°C at ~38ka cal to 27.4±3.8°C during the temperature
minimum at ~45–43 kacal, compared to the modern-day tempera-
ture seasonality of 19±2.9°C.
Relationship with human presence
To test the temporal overlap between the equid specimens yield-
ing the isotopic climate data and the presence of H. sapiens we used
χ2 tests and agreement indices of the OxCal Combine function for
groups of direct dated equids, H. sapiens remains and anthropo-
genically modified faunal bone fragments. A table of all test results
can be found in the associated online supplementary material at
https://osf.io/wunfd/.
The direct dates of all equid individuals overlapping with the age
ranges of the LRJ deposits (~48–43 kacal) are statistically indistuin-
guishable from at least one directly dated H. sapiens fragment and, in
many cases, also from at least one anthropogenically modified faunal
bone fragment. Importantly, this includes the equids that yielded
the lowest δ18O values (R10124, ETH-111922 and R10126, ETH-111920),
which show a calibrated date range of 45,000–43,100cal (Fig. 3).
The direct date of R10126 is statistically indistinguishable (χ2=0.170,
d.f.=2, (5% 5.991), Acomb=117.7) to those of a H. sapiens fragment
(R10875, ETH-127625) and a cut-marked bone from layer 8 (16/116-
159091, ETH-118367), while the date of R10124 is statistically indis-
tinguishable (χ
2
=0.079, d.f.=3, (5% 7.815), Acomb=194.5) from two
H. sapiens fragments (R10879, ETH-127628 and R10396 ETH-115246)
and an equid fragment with percussion notches (16/116-159318, ETH-
111935). Other H. sapiens specimens and anthropogenically modified
Modern reference
Modern reference
Modern reference
Summer Mean annual Winter
34 36 38 40 42 44 46 48 34 36 38 40 42 44 46 48 34 36 38 40 42 44 46 48
−20
−10
0
10
20
1 ka cal BP
Estimated air temperature (°C)
Fig. 2 | Air temperature estimates derived from δ18O measurements generally
fall below modern-day conditions. Lowest temperatures are observed in
the ~45–43 ka cal interval, where they fall ~7–15°C below modern day and
mean annual temperatures below freezing. Oxygen isotope data from several
individuals were grouped into time bins according to clusters of radiocarbon
dates and δ18O measurements (Supplementary Table 6). Plotted points represent
temperature estimates for each time bin. Error bars represent combined
uncertainty for each temperature estimate, taking into account the uncertainty
of each temperature calibration step (Supplementary Text 5). Ndatapoints for
each error bar varies by season and time bin and can be found in detail in
Supplementary Table 6. In the time bins δ18Odw estimates are based on a variable
number of tooth specimens with n36–39 ka=7, n42–43 ka=3, n43–45 ka=2 and n45–48 ka=3.
Summer and winter temperatures were estimated from inverse modelled δ18O
time series, while annual means were derived directly from unmodelled δ18O
(Supplementary Text 5). Lines and shaded ribbons of modern comparative data
represent means and one standard deviation of modern climate observations
(MAT, Tcoldest month, Twarmest month) for 1961–2009 from the ClimateEU model59).
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bone fragments from the LRJ deposits date to a slightly earlier period
~48–45ka cal coinciding with equid specimens yielding tempera-
tures that fall ~3–8°C lower than today but are less extreme than those
from the ~45–43 kacal interval (Fig. 3).
Comparison of the uncalibrated dates confirms that the chrono-
logical overlap between H. sapiens specimens and the coldest phase at
~45–43 kacal is independent of the calibration curve used (Supple-
mentary Fig. 13). The two equids yielding lowest temperature data (ETH-
111922 41,490±36014C and ETH-111920 40,740±33014C) overlap
with the dates of seven of ten human specimens (Supplementary
Fig. 13) and are almost identical to the ages of R10879 (41,429±76514C)
and R10396 (41,570±42014C).
Taxonomic identifications of bone fragments with anthropogenic
surface modifications from layers 9 and 8 show that they predomi-
nantly originate from reindeer but also include equid fragments. While
the total number of fragments (n=12) is too low to make robust infer-
ences about taxonomic representation of hunted herbivores, this does
indicate that some equids did overlap with H. sapiens occupations and
that there is no immediate indication of a systematic difference in the
prey taxa targeted by H. sapiens compared to carnivores15.
GS 12
GI 12
GS 13
−43
−41
−39
−37
δ18O (‰ VSMOW)
Per cent arboreal
Per cent Betula
0
20
40
60
Pollen (%)
10
12
14
δ18Ophos (‰ VSMOW)
Layer age ranges
IX−XI
9
8
7
34,000 36,000 38,000 40,000 42,000 44,000 46,000 48,000 50,000
Years cal BP
Layer
Anthropogenic modified bone H. sapiens Excavation 1932−1938 2016−2022
Fig. 3 | H. sapiens presence coincides with the coldest temperatures
documented by equid δ18O data. Comparison of equid δ18O data (top) with
directly dated H. sapiens remains 13 (bottom, turquoise symbols) demonstrates
extensive overlap of H. sapiens presence with the coldest temperatures
documented between ~45 and 43 kacal (marked by blue shading). This
coldest, low δ18O phase overlaps with the age ranges of both the LRJ layer 8
and the beginning of undiagnostic layer 7 (ref. 13) (modelled 95% probability
layer age ranges of the MPI-EVA/TLDA excavation in purple) but the direct
dates of H. sapiens remain and faunal bone fragments with anthropogenic
surface modifications clearly show that they endured the cold subarctic steppe
conditions evidenced by the stable isotope data at this time. This also holds
true independent of the calibration curve, as seen in the uncalibrated dates
(Supplementary Fig. 13). Top panels show the relevant Greenland stadials (GS)
and interstadials (GI) recorded in the NGRIP ice cores60 and the proportions of
total arboreal pollen (dark brown) and Betula pollen (ochre) in the Füramoos
pollen record from southwestern Germany29,61. Data are presented as mean±95%
calibrated age ranges (n=1 bone or tooth enamel sample for each data point),
while point shape indicates whether specimens were found in the Hülle (1932–
1938, circles) or the TLDA/MPI-EVA (2016–2022, triangles) excavation collection.
We argue that H. sapiens fragments from the Hülle collection (labelled IX–XI
here) all originate from the LRJ deposits (layer X) and were sometimes assigned
to a mixture of layer X and adjacent strata by the original excavators due to rough
excavation methods (details in ref. 13). We have pooled all these samples here
to reflect this. Credits: equid silhouette by Mercedes Yrayzoz, vectorized by T.
Michael Keesey (PhyloPic); human silhouette from NASA Pioneer plaque.
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Discussion
Using multiple isotope analyses of directly
14
C-dated equid teeth com-
pared to directly dated H. sapiens fossils and an updated site chro-
nology, we provide evidence that H. sapiens associated with the LRJ
occupations of Ranis were present in central Europe during subarctic
climatic conditions in a cold open environment, probably including
a severe cold episode at ~45–43 kacal. This shows that H. sapiens
successfully operated in harsh environmental conditions during an
early northward range expansion into central Europe.
Equid δ18Ophos data reported here are among the lowest ever
reported in Europe for MIS5 to MIS3 (δ
18
O
mean annual
=9–13.5‰ for the
LRJ). Direct comparisons are limited to data from other equids, where
most MIS3 data from Germany do not fall below 14‰ (ref. 16). Examples
with low values from Late Pleistocene stadial contexts in Germany
(Bocksteinhöhle, Vogelherdhöhle and Villa Senckendorff
17,18
) and in
Switzerland (Boncourt Grand Combe, MIS3, Courtedoux-Va Tche
Tcha, MIS5a, ref. 19) range from 12.1‰ to 13.2‰ for mean annual data.
This is similar to the higher values from Ranis observed for ~48–45
kacal and ~43–39 kacal. Data from Ranis for ~45–43 kacal,
however, descend 1.5–2‰ lower than this (Fig. 1). The data from ~45–43
kacal fall ~0.5–1.5‰ lower than even the lowest equid δ18Ophos data
reported from the Initial Upper Palaeolithic occupation of Bacho Kiro
Cave (minimum δ18Omean annual=11.2‰), Bulgaria, which has been used
to reconstruct subarctic climatic conditions for an early presence of H.
sapiens in southeastern Europe
9
. Reconstructed drinking water δ
18
O
dw
comparisons with other species suggest closest matches with Late
Pleistocene data in Scandinavia and Russia, including data from the Last
Glacial Maximum20,21. On the basis of characteristics of the study area
and on the presence of a sinusoidal signal, we argue that δ18Ophos data
reflect palaeotemperatures (Supplementary Text 2), thus our results
demonstrate low temperatures, some of them remarkably so. Palae-
otemperature estimates show a variability of conditions with a cooling
trend but even the comparatively warmer episode at ~48–45 kacal
shows mean annual temperatures of <5°C and based on δ18Oprecip most
closely matches current climatic conditions in subarctic climate zones
in northern Finland (for example, GNIP station Rovaniemi22). Mean
annual temperatures then descend even further to below freezing for
the coldest interval at ~45–43 kacal. For this interval, our results
correspond to temperature anomalies of ~7–15°C below modern-day
conditions with largest anomalies in winter (temperature estimation
assumptions in Supplementary Text 2). This is paired with a very strong
temperature seasonality of up to 27±5°C, indicating a continental
cold subarctic to tundra climate with closest modern-day matches
in northwestern Russia (for example, GNIP stations in Amderma and
Pechora
22
). Importantly, summer temperature estimates in some cases
fall below the 12°C warmest month isotherm that dictates the Eurasian
northern tree line23, indicating that tree growth may have been impos-
sible in some of the climatic phases captured here. Air temperatures of
~10–15°C below modern day are consistent with full stadial conditions
and have been reconstructed for particularly severe Greenland stadials
(GS) in central Europe24–27.
The cooling trend from ~48 to 44 kacal into full stadial con-
ditions followed by a rapid temperature increase after ~44ka cal
matches well with the documented slow cooling and rapid warming of
Dansgaard–Oeschger (DO) events and based on tentative correlations
with long-term climatic records may capture a Greenland interstadial
(GI) to GS transition culminating in a pronounced cold phase such as
GS12 or GS13 (refs. 27–30). While an assignment to a specific DO event
may not be possible because of the chronometric dating uncertain-
ties involved, the comparison suggests that the equid δ18O record
captures millenial-scale climatic variability that occurred during the
time of the LRJ.
Remarkably high δ15N values of equid dentine and mandible bone
collagen in the ~45–43 kacal interval (~7–9‰) compared to earlier
and later equid data suggest either a hypergrazer feeding ecology or
dry soil conditions during this interval. Nitrogen isotope variability
in arctic biomes shows the highest values in grasses and herbaceous
plants over shrubs or trees and this transfers to high δ
15
N values in
specialized grazers
31
. Glacial phases are often accompanied by low
δ
15
N values in fauna due to limited nitrogen availability and reduced
bacterial activity in cold-wet soils, while high δ
15
N values are observed
in phases with higher temperatures or low moisture availability
32,33
.
As δ
18
O values demonstrate low temperatures in this interval, high
δ
15
N values could indicate dry soil conditions and/or strong grazing
specialization of equids, which both imply the presence of an open
steppe environment. This matches with reconstructions of grass steppe
environments in central Europe during MIS3 stadials30,34. Equid dentine
and mandible bone collagen δ
13
C values are consistent with feeding in
an open grassland environment35 and the lack of diachronic change is
in line with relatively small climatic impacts on δ13C of C3 plants com-
mon to Pleistocene European biomes.
Zinc stable isotope values of the food web at Ranis follow expected
trophic level relationships with low values in carnivores and high values
in herbivores36,37. Within herbivores, δ66Zn seems to reproduce a pat-
tern of higher values in taxa commonly consuming more grass (equids)
and lower values in typical browsers to mixed feeders such as Cervus
elaphus (Extended Data Fig. 2). A similar pattern has been observed
in a few European and African food webs and agrees well with higher
δ
66
Zn values observed in low-growing plants over higher-standing tree
or shrub leaves
37–39
but its robustness is still debated
36
(Supplementary
Text 4). While the Ranis data cannot be used to definitively confirm this
idea, they are tentatively consistent with it. If true, particularly high and
seasonally invariant δ
66
Zn observed in equids in the ~45–43ka cal
cold interval would support an interpretation of a hypergrazer feed-
ing niche of equids in an open steppe environment. The statistically
significant correlation of δ66Zn with δ15N (Supplementary Fig. 12) is also
consistent with both proxies being driven by grass consumption but
due to the effects of soil nutrient cycling on both isotopic systems
40
, a
relationship with dry soils and consequent changes in soil biochemical
cycles is also possible.
Cold temperature conditions and an open grassland or tundra
environment for the LRJ at Ranis match the faunal spectrum which
includes cold-adapted fauna such as wolverine (Gulo gulo), reindeer
(Rangifer tarandus), woolly mammoth (Mammuthus primigenius)
and woolly rhinoceros (Coelodonta antiquitatis), with reindeer being
the predominant herbivore taxon15. Furthermore, sedimentological
analyses suggest a drop in temperature from layer 9 to the start of layer
7 based on a pronounced decrease in organic carbon and total nitrogen
content
13
, lending support to the decreasing temperatures from ~48
to 43 kacal reported here.
Our results show that climatic conditions throughout the LRJ
occupations, even during the earliest phase ~48–45 kacal, were
characterized by temperatures substantially below modern-day condi-
tions. Although a direct contextual connection through anthropogenic
modification cannot be established, the chronological overlap between
the direct dates of H. sapiens remains and anthropogenically modified
bone fragments with those of the equid individuals that yielded low
temperature results indicates that H. sapiens faced subarctic to tundra
climatic conditions, probably even those of the severe cold climatic
phase 45,000–43,000cal. Zooarchaeological analysis suggests
that this presence was characterized by ephemeral occupations, either
due to short occupation, task-specific site use or small group sizes
15
,
although the direct radiocarbon dates of H. sapiens remains suggest
intermittent site visits across at least a thousand years. Such a site-use
pattern, perhaps in the context of frequent movements between sites,
may have been a response to the subarctic steppe environment recon-
structed for the LRJ. Micromorphological evidence for increased fire
use in layer 8 compared to layers 9 or 7 (ref. 13) could also be indicative
of a behavioural adaptation to the cooling climate. Owing to the few
human modifications on faunal remains, we cannot determine the
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seasonality of site occupations by LRJ H. sapiens groups
15
. However,
more long-term palaeoclimatic records and limited evidence from
other LRJ sites (see below) indicate that a subarctic steppe or tundra
landscape would have extended over large areas in central Europe,
where human groups would have faced similar conditions during
potential seasonal movements. The ephemeral H. sapiens presence
at the site implies that the climatic data probably also cover phases of
site formation where humans were absent but we argue that the tem-
poral overlap between directly dated equids, H. sapiens remains and
anthropogenically modified bones and the reflection of millenial-scale
climatic variability in our record does suggest that we can broadly
characterize the climates faced by LRJ H. sapiens as cold to very cold
during most of the LRJ formation period.
The association of H. sapiens with the LRJ suggests a rapid range
expansion across the northern European plain as far as the British
Isles
13
, which may have been enabled by the resilience to cold condi-
tions and success in steppe environments documented here. Direct
environmental evidence from other LRJ sites is sparse. Nonetheless,
the association of LRJ material with cold-adapted fauna at Grange Farm,
United Kingdom, and Schmähingen, Germany, as well as biomarker
and pollen evidence for climate cooling and open landscapes during
the Jerzmanowician occupation of Koziarnia Cave, Poland, suggests
that association with cold climatic conditions may be a more com-
mon feature of the LRJ than previously noted
41–43
. Genetic data and
technological analyses allude to a potential connection between the
Ranis LRJ to populations and technocomplexes further east13, includ-
ing potentially the Initial Upper Palaeolithic44, which is associated
with a cold-climate H. sapiens presence at Bacho Kiro Cave, Bulgaria9.
Cold-steppe environments that provided open landscapes and sup-
ported large herds of prey fauna may have actively supported a rapid
dispersal of these connected populations across the northern and
eastern European Plain
45,46
. At the same time, our study joins increasing
recent evidence for a more complex patchwork of early dispersals of
our species in different periods and in more diverse ecological settings
than previously appreciated
1–3,5,9,10
, raising the question of whether
pioneering groups of H. sapiens may not be more accurately described
as climatically resilient generalists.
Methods
Study design, materials and sampling
A multi-isotope study design was chosen to reconstruct a variety of
climatic, environmental and ecological aspects of the LRJ and Upper
Palaeolithic deposits of Ilsenhöhle in Ranis. A multi-isotope approach
also has substantial benefits in reducing equifinality in the interpreta-
tion of each isotopic proxy. Oxygen stable isotope analysis was chosen
as the main palaeoclimatic proxy, while strontium isotope analysis
serves to confirm that sampled animals did not undergo long-distance
migrations that could affect the δ
18
O signal (Supplementary Texts 2
and 3). Carbon and nitrogen stable isotope analyses were conducted
to reconstruct dietary ecology, the structure of the plant biome and
water availability in the past. We add to this aspect using zinc stable
isotope analysis, a non-traditional stable isotope proxy with potential
to elucidate herbivore dietary ecology (Supplementary Text 4).
A total of 16 equid teeth were selected for stable isotope analysis
and radiocarbon dating (Supplementary Table 1). Only fully formed
and mineralized teeth were considered and first molars (M1) were
excluded to prevent the influence of mother’s milk consumption on
oxygen isotope ratios. Identification of tooth position was achieved
with the help of an experienced equid tooth specialist and teeth where
an M1 identification could not be confidently excluded were not sam-
pled. Specimens were obtained both from the collections of the 1930s
excavation by W. Hülle (sample numbers starting with ‘R’, n=14) and
the 2016–2022 excavation by the TLDA and the MPI-EVA (sample num-
bers starting with ‘16/116’, n=2). Teeth were chosen to obtain data
on the lower part of the depositional sequence from the black layer
(TLDA/MPI-EVA: 6 black, Hülle VIII) downward including the LRJ occu-
pations and adjacent layers (Extended Data Fig. 1) and sequentially
sampled to yield subannually resolved stable isotope data (Supple
-
mentary Text 5). In the Hülle excavation, this encompasses layers VIII
(Schwarze Schicht), IX (Mittlere Braune Schicht), X (Graue Schicht)
and XI (Untere Braune Schicht), where layer X is associated with the LRJ
technocomplex. These layers correspond to the depositional sequence
from layer 6 black to layer 14 in the TLDA/MPI-EVA excavation, where
the LRJ is associated with layers 8 and 9 (ref. 13). Layer information in the
Hülle faunal collection is recorded using deposit colour (for example,
‘Graue Schicht’, meaning grey layer) and approximate depth. For the
two brown layers identified by Hülle in the lower depositional sequence
(IX and XI), these labels can include colour variations (for example,
Braune Schicht, Schokobraune Schicht and Rotbraune Schicht), which
probably reflect stratigraphic information but also colour differences
between site areas, while layer positions (for example, ‘mittlere’ and
‘untere’, meaning middle and lower) are almost always omitted. Owing
to sloping terrain and compression of deposits by rockfall in some
areas of the site13, depth information is often of limited use in assigning
layer designations. Colour and depth descriptions were used to assign
layer association as best as possible but all equid specimens were also
directly radiocarbon dated using dentine or mandible bone collagen
samples to confirm their chronological position.
Because of the predominant role of carnivores in accumulating the
faunal remains found in the Ranis LRJ and Upper Palaeolithic deposits
and the palimpsest nature of the deposits, the link between faunal
stable isotope data and H. sapiens activity at the site is less direct than
for sites where the faunal assemblage is predominantly anthropo-
genically accumulated. Moreover, we rely on a comparison of direct
radiocarbon dates of the equids analysed for stable isotopes with
those of the archaeological layer boundaries, direct dates of H. sapiens
skeletal remains and anthropogenically modified faunal fragments to
establish the archaeological context for the equid remains sourced
from the Hülle collection. This approach, while unavoidable due to
the characteristics of the site, carries some uncertainty in relating the
isotopic climate evidence to periods of H. sapiens site occupation.
Indeed, it is most likely that the climatic data generated in this study
cover both periods of H. sapiens presence at Ranis and periods where
humans were absent. We use χ2 tests and agreement indices of the OxCal
Combine function to test the chronological agreement between the
direct dates of the equid specimens yielding the climatic data on the
direct dates of H. sapiens remains and anthropogenically modified
bones from LRJ contexts to test the probability of a link with H. sapiens
presence as best as possible.
Samples of ~300–600mg of dentine or mandible bone were
obtained for collagen extraction from tooth roots when available. If
roots were not preserved, pieces were cut from lower sections of the
tooth crown and tooth enamel was mechanically removed before dem-
ineralization. For two specimens (16/116-123510 and 16/116-124286),
adhering mandibular bone was available and sampled instead of tooth
dentine. Sample pieces were removed using a diamond-coated rotary
disk after cleaning of surfaces using air abrasion (Supplementary Text 5).
In addition to the equid specimens chosen for δ18O, δ13C, δ15N, δ66Zn
and 87Sr/86Sr analysis, a total of 24 tooth enamel specimens representing
a variety of herbivore, omnivore and carnivore taxa were chosen for
further δ
66
Zn and
87
Sr/
86
Sr analyses to explore patterns across the food
web (Supplementary Table 1). These specimens were obtained from the
Hülle collection from contexts thought to correspond to the brown
layer IX. Given the documentation of the finds from the brown layers
in the collection described above, this sample probably represents to
some degree a mix of specimens of different LRJ and Upper Palaeolithic
stratigraphic units (Supplementary Text 1 and Supplementary Fig. 1)
dating between ~48 and 36 kacal. While less than ideal, analysis of
the directly dated equid specimens shows that diachronic changes
in δ
66
Zn are too small to affect broader dietary patterns and trophic
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Article https://doi.org/10.1038/s41559-023-02318-z
relationships in the food web (Supplementary Text 4). Tooth enamel
samples from these specimens were obtained either as powder or piece
samples in a positive pressure Flowbox following methods described
in Supplementary Text 5. In some cases, several teeth from the same
mandible were sampled to obtain sufficient tooth enamel.
Oxygen stable isotope analysis
Tooth enamel powder samples were converted to silver phosphate
for oxygen isotope analysis of bioapatite phosphate using digestion
with hydrofluoric acid, followed by crash precipitation of silver phos-
phate47,48 (Supplementary Text 5). Following recommendations in ref. 49
we did not use an oxidative pretreatment before silver phosphate
preparation. Oxygen isotope delta measurements of Ag
3
PO
4
were con-
ducted in triplicate using a high-temperature elemental analyser (TC/
EA) coupled to a Delta V isotope ratio mass spectrometer via a Conflo IV
interface (Thermo Fisher Scientific) (Supplementary Text 5). Oxygen
isotope delta values were two-point scale normalized to the VSMOW
scale using matrix-matched standards calibrated to international
reference materials and scale normalization was checked using three
separate quality control standards. Details of normalization standards
and quality control outcomes can be found in Supplementary Text 5.
Average reproducibility of sample replicate measurements was 0.25‰.
Inverse modelling and palaeotemperature estimation
Before seasonal palaeotemperature estimation an inverse model fol-
lowing ref. 50 was applied to sinusoidal δ18Ophos time series to remove
the time averaging and amplitude damping effects caused by the
extended nature of tooth enamel mineralization and the sampling
procedure (Supplementary Text 5). It should be noted that this inverse
model does not account for the successive decrease in tooth growth
and mineralization speed that is known to occur in horses towards the
completion of tooth formation. Seasonal amplitudes reconstructed
here, therefore, probably represent minimum amplitudes (Supple-
mentary Text 5). Following recommendations developed in ref. 9,
summer peak and winter trough values were obtained from the inverse
modelled δ
18
O curves and processed for palaeotemperature estima-
tion, while mean annual temperatures were estimated on the basis of
unmodelled annual means. Summer peak and winter trough values
were first identified by visual inspection in the original unmodelled
δ
18
O time series (Supplementary Fig. 2) and corresponding areas on
the inverse model outcome were used to yield corrected summer δ
18
O
and winter δ
18
O values. Unmodelled annual means were calculated
as the mean of unmodelled summer peak and winter trough values
following ref. 9. Detailed information on the modelling procedure is
provided in Supplementary Text 5 and we provide all associated code
and data, including the parameters used for each model run and the
model outcomes for all specimens of this study in the associated online
repository at https://osf.io/wunfd/.
Following methods in ref. 51, air temperature estimates were
derived via two regression steps using the empirically determined
relationships between (1) tooth enamel δ
18
O
phos
and drinking water δ
18
O
(δ
18
O
dw
) and (2) δ
18
O of precipitation (δ
18
O
precip
) and air temperature.
Regression relationships were established using modern calibration
datasets of equid tooth enamel δ
18
O and drinking water δ
18
O as well
as temperature and δ18Oprecip data from meteorological measurement
stations. Details on modern calibration datasets and the conversion
procedure are described in Supplementary Text 5. All calculations with
the specific conversion equations can be reproduced using the data
and code provided in the associated online repository at https://osf.
io/wunfd/. Printed conversion equations based on the same data and
excel files to conduct equivalent conversions are also available in ref. 9.
To confirm that herbivore δ18O values reflect climatic influences
without ecological or behavioural biases, some studies recommend
the use of several taxa from the same archaeological units. Owing to
a lack of teeth from other taxa suitable for sequential sampling (for
example, large bovids) this was unfortunately not possible in this
study (Supplementary Text 2). However, equid δ
18
O
dw
values have been
shown to be in excellent agreement with those of other sympatric taxa
(Supplementary Text 2).
Collagen extraction and radiocarbon dating
Collagen was extracted from the equid teeth in the Department of
Human Evolution at the MPI-EVA using HCl demineralization, NaOH
humic acid removal, gelatinization and ultrafiltration steps, follow-
ing the protocol in refs. 52,53. (Supplementary Text 5). The suitability
of the extracts for dating was assessed on the basis of collagen yield
(minimum requirement ~1%) and the elemental values54, as reported
in Supplementary Text 5 and Supplementary Table 2. All extracts were
characteristic of well-preserved collagen (Supplementary Table 2)
so were submitted for
14
C dating via accelerator mass spectrometry.
Three of 16 collagen extracts were graphitized and dated at the Curt
Engelhorn Center for Archaeometry gGmbH (CEZA, laboratory code:
MAMS), while the remaining 13 extracts were dated at the Laboratory
for Ion Beam Physics at ETH Zurich, Switzerland (laboratory code: ETH;
Supplementary Text 5). Aliquots of a background bone (>50,000)
were pretreated and dated alongside the equid samples to monitor
laboratory-based contamination and were used in the age calculation
of the samples. The 14C dates were calibrated in OxCal 4.4 (ref. 55) using
the IntCal20 calibration curve56. Uncalibrated 14C dates (14C ) are
reported with their 1σ error and, in the text, calibrated ranges (cal)
are reported at the 2σ range (95% probability). Calibrated ages at the
1σ range (68% probability) can be found in Supplementary Table 2. All
dates have been rounded to the nearest 10years.
Carbon and nitrogen stable isotope analysis
Subsamples of collagen extracts were analysed for their elemental com
-
position (%C, %N) and carbon and nitrogen stable isotope composition
using a Flash 2000 Organic Elemental Analyser coupled to a Delta XP
isotope ratio mass spectrometer via a Conflo III interface (Thermo Fisher
Scientific; Supplementary Text 5). Samples were analysed in duplicate
and stable isotope delta values were two-point scale normalized using
international reference material IAEA-CH-6, IAEA-CH-7, IAEA-N-1 and
IAEA-N-2 for δ13C and δ15N, respectively. Two inhouse quality control
standards were used to check scale normalization and evaluate analyti-
cal precision. Replicate sample measurements and measurements of
the in-house standards indicate a measurement precision of 0.1‰ or
better for both δ13C and δ15N. Details of calibration methods and qual-
ity control indicators can be found in Supplementary Text 5. Elemental
composition and C/N ratios used for quality checks of collagen integrity
are reported in Supplementary Text 5 and Supplementary Table 2.
Strontium and zinc stable isotope analysis
Zinc and strontium extraction were both conducted on ~10mg of
tooth enamel powder or pieces. In the case of sequentially sampled
equid teeth, two subsamples per specimen, representing the summer
and winter seasons (based on δ18O sinusoid peaks and troughs), were
selected for δ66Zn and 87Sr/86Sr analysis. Samples were processed using
standard acid digestion and column chromatography purification
protocols following refs. 37,57,58. (Supplementary Text 5). Isotopic
measurements were conducted using a Neptune Multi-Collector Induc-
tively Coupled Plasma Mass Spectrometer (MC-ICPMS, Thermo Fisher
Scientific). Procedural blanks and aliquots of quality control standards
were processed alongside each batch of samples to quality check wet
chemistry and isotope measurement quality. Details of the instru-
mental setup, scale normalization and quality control indicators can
be found in Supplementary Text 5. A subset of samples was analysed
in duplicate with an average reproducibility of 0.000008 for 87Sr/86Sr
and 0.01‰ for δ66Zn.
It has been shown that dental enamel reliably preserves biogenic
strontium and zinc isotope values (see Supplementary Text 4 for details
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Article https://doi.org/10.1038/s41559-023-02318-z
on the preservation of δ66Zn in fossil tooth enamel). As an additional
diagenetic check we evaluated isotope measurements against elemen-
tal concentration data to confirm the preservation of biogenic isotopic
ratios. A lack of relationship between the two in our results indicates
that incorporation of diagenetic Zn or Sr is unlikely (Supplementary
Figs. 14 and 15).
Reporting summary
Further information on research design is available in the Nature Port-
folio Reporting Summary linked to this article.
Data availability
All data presented in this study are openly accessible in electronic form
in an Open Science Framework repository (https://doi.org/10.17605/
OSF.IO/WUNFD) at https://osf.io/wunfd/ and stable isotope data will
be deposited in the IsoArch database (https://isoarch.eu/). Data avail-
able in the OSF repository include all stable isotope measurements,
radiocarbon dates and isotope-derived palaeotemperature estimates.
Code availability
R code used to conduct the analyses and reproduce the manuscript
and Supplementary Information of this study can be accessed in an
Open Science Framework repository (https://doi.org/10.17605/OSF.
IO/WUNFD) at https://osf.io/wunfd/. This includes code and data to
reproduce the oxygen isotope inverse modelling procedure and the
palaeotemperature estimations. More detailed documentation and
example scripts and files for the inverse model and the temperature
estimation can be found at https://github.com/scpederzani/Oxygen_
Inverse_Model and https://github.com/scpederzani/Isotope_Tem-
perature_Calibration. The manuscript and Supplementary Information
were written using R and Quarto so that figures and analyses can be
transparently reproduced using the available code and data. Details
of R packages and computing environment can be found in Supple-
mentary Text 5.
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Acknowledgements
The re-excavation of Ilsenhöhle in Ranis was conducted by the TLDA
and the MPI-EVA. We thank the TLDA and the State Oice for Heritage
Management and Archaeology Saxony-Anhalt—State Museum of
Prehistory (LDA) for the opportunity to study the Ranis faunal material.
In particular, we thank R. Hülsho (LDA) and I. Widany (LDA) for
assistance in accessing the LDA collections. We thank M. Kaniecki,
L. Klausnitzer, S. Hesse and P. Dittmann (MPI-EVA) for technical
assistance during stable isotope and radiocarbon sample preparation.
S. Steinbrenner is thanked for technical assistance with thermal
conversion elemental analysis isotope ratio mass spectrometry (TC/
EA)-IRMS maintenance and EA-IRMS measurements. Thanks are
also due to E. Schulz-Kornas (University of Leipzig) for assistance
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Nature Ecology & Evolution | Volume 8 | March 2024 | 578–588 588
Article https://doi.org/10.1038/s41559-023-02318-z
with identifying equid tooth positions and S. Tüpke (MPI-EVA) for
conducting high-resolution photography of equid tooth specimens.
We express our gratitude to J. Krause (MPI-EVA) for support during
the H. sapiens specimen identiication work conducted by H.R.,
J.O. and H.D. The stable isotope work and radiocarbon dating was
funded by the Max Planck Society as part of the PhD and postdoctoral
project of S.P. and by the German Research Foundation (DFG) as
part of the PALÄODIET Project (378496604) awarded to K.J. and T.T.
S.P. is supported by a German Academy of Sciences Leopoldina
postdoctoral fellowship (LPDS 2021-13). K.B. is supported by a Philip
Leverhulme Prize from The Leverhulme Trust (PLP-2019-284). G.M.S.
received funding from the European Union Horizon Europe Research
and Innovation Programme under Marie Skłodowska–Curie Grant
Agreement 101027850. J.M. received funding from the DFG (Project
505905610). D.M. received funding from the European Union’s
Horizon 2020 research and innovation programme under the Marie
Skłodowska-Curie Grant Agreement 861389 - PUSHH.
Author contributions
The study was designed by S.P., K.B., M.W., S.P.M., J.-J.H., K.J. and T.T.
Archaeological excavation was undertaken by M.W., T.S. and S.P.M.,
who all contributed contextual information. Zooarchaeological and
palaeontological analyses were performed by G.M.S. Radiocarbon
dating and carbon and nitrogen stable isotope analysis were
conducted by H.F. and S.T. Geological and sedimentological analyses
were conducted by A.K. and T.L. Characterization and dating of the H.
sapiens remains was carried out by D.M., H.F., E.I.Z., V.S.M., K.R., F.W.,
H.R., J.O. and H.D. T.S., H.-J.D., H.D. and H.M. provided study specimens
and contextual information. Sampling, sample processing for oxygen
stable isotope analysis and TC/EA-IRMS analysis were carried out
by S.P. Sampling for strontium and zinc stable isotope analysis were
conducted by S.P. and M.T. Sample processing for strontium and zinc
stable isotope analysis was carried out by M.T. MC-ICPMS analysis
was conducted by N.B. and J.M. Code and data analyses were written
and conducted by S.P. M.L.C. contributed to the spatial analysis of
strontium stable isotope results. S.P. wrote the paper with input from
all authors.
Funding
Open access funding provided by Max Planck Society.
Competing interests
The authors declare no competing interests.
Additional information
Extended data is available for this paper at
https://doi.org/10.1038/s41559-023-02318-z.
Supplementary information The online version contains supplementary
material available at https://doi.org/10.1038/s41559-023-02318-z.
Correspondence and requests for materials should be addressed to
Sarah Pederzani.
Peer review information Nature Ecology & Evolution thanks William
Banks, Christophe Lécuyer, Trine Kellberg Nielsen and the other,
anonymous, reviewer(s) for their contribution to the peer review of
this work. Peer reviewer reports are available. Peer reviewer reports
are available.
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© The Author(s) 2024
1Department of Human Evolution, Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany. 2Archaeological Micromorphology and
Biomarkers Laboratory (AMBI Lab), Instituto Universitario de Bio-Orgánica Antonio González, Universidad de La Laguna, San Cristóbal de La Laguna,
Spain. 3Department of Archaeology, University of Aberdeen, Aberdeen, UK. 4Ancient Genomics Lab, The Francis Crick Institute, London, UK. 5isoTROPIC
Research Group, Max Planck Institute for Geoanthropology, Jena, Germany. 6Institute of Geosciences, Goethe University Frankfurt, Frankfurt, Germany.
7Géosciences Environnement Toulouse, Observatoire Midi Pyrénées, UMR 5563, CNRS, Toulouse, France. 8State Ofice for Heritage Management
and Archaeology Saxony-Anhalt—State Museum of Prehistory, Halle, Germany. 9State Authority for Mining, Energy and Geology of Lower Saxony
(LBEG), Hannover, Germany. 10Terrestrial Sedimentology, Department of Geosciences, University of Tübingen, Tübingen, Germany. 11CNRS, UMR 7209
Archéozoologie et Archéobotanique—Sociétés, Pratiques et Environnements (MNHN-CNRS), Paris, France. 12Department of Human Origins, Max Planck
Institute for Evolutionary Anthropology, Leipzig, Germany. 13Chair of Paleoanthropology, CIRB (UMR 7241—U1050), Collège de France, Paris, France.
14Department of Anthropology, California State University Northridge, Northridge, CA, USA. 15Department of Archaeogenetics, Max Planck Institute
for Evolutionary Anthropology, Leipzig, Germany. 16Thuringian State Ofice for the Preservation of Historical Monuments and Archaeology, Weimar,
Germany. 17University of Bordeaux, CNRS, Ministère de la Culture, PACEA, UMR 5199, Pessac, France. 18School of Anthropology and Conservation,
University of Kent, Canterbury, UK. 19Department of Chemistry G. Ciamician, Alma Mater Studiorum, University of Bologna, Bologna, Italy. 20Applied
and Analytical Palaeontology, Institute of Geosciences, Johannes Gutenberg University, Mainz, Germany. 21Globe Institute, University of Copenhagen,
Copenhagen, Denmark. 22Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany. 23Department of
Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, USA. 24Friedrich-Alexander-Universität Erlangen-Nürnberg, Institut für Ur- und
Frühgeschichte, Erlangen, Germany. e-mail: scpederz@ull.edu.es
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Article https://doi.org/10.1038/s41559-023-02318-z
Extended Data Fig. 1 | Location and stratigraphy of Ilsenhöhle in Ranis,
Germany. A - Location of Ilsenhöhle in Ranis, Thuringia, central Germany.
B - Hydrotopographical setting of the area surrounding Ranis in the Orla valley.
The Thuringian highlands can be seen to the south of the site. Other notable
rivers include the Saale River passing south and west of Ranis. Elevation contour
lines are spaced 200m apart. Elevation data from the European Digital Elevation
Model version 1.162. Waterways imported from OpenStreetMap63.
C - Schematic stratigraphy of the 1930s Hülle excavation and the 2016–2022
TLDA/MPI-EVA excavation with layer correlations (layer numbers in circles).
Samples analysed here roughly cover the time period from the Lincombian–
Ranisian-Jerzmanowician (LRJ) Layers 8 and 9 marked in red to the Upper
Palaeolithic occupation of Layer 6. Rockfall events are marked as ‘R’.
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Nature Ecology & Evolution
Article https://doi.org/10.1038/s41559-023-02318-z
Extended Data Fig. 2 | Zinc stable isotope values across the Ranis food web.
Zinc stable isotope ratios of a range of taxa show typical trophic relationships
with low δ66Zn values for carnivores, high values for herbivores and intermediate
omnivores and bone eating carnivores. Herbivores show a pattern with highest
δ66Zn values in equids and lower values in typical browser to mixed feeding cervid
taxa such as Cervus elaphus. This is potentially consistent with grazer-browser
δ66Zn patterns observed in a European Pleistocene food web38 and a modern
African food web37 and with limited studies of different plant parts39. If
confirmed, this suggests that higher δ66Zn in some equids could be due to a
hypergrazer feeding ecology (see Supplementary Text 4). Shapes indicate stages
of tooth development, as teeth formed during nursing or in utero can exhibit
higher δ66Zn values (see Supplementary Text 4). It should be noted that for equids
summer and winter δ66Zn values are plotted (2 per tooth), while other taxa are
represented by one measurement per specimen.
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Nature Ecology & Evolution
Article https://doi.org/10.1038/s41559-023-02318-z
Extended Data Fig. 3 | Summer, winter and mean annual estimates of
environmental water oxygen isotope values compared to modern meteoric
water sources. Reconstructed oxygen isotope composition of drinking water
(δ18Odw) fall substantially below δ18O of modern-day water sources (precipitation
- solid ribbons; rivers - hatched ribbons; springs - dotted ribbons), particularly
for mean annual and winter values. This indicates that temperatures were
substantially below modern-day conditions. Summer δ18Odw reconstructions
partially overlap with modern spring δ18O due to the pronounced seasonal
buffering in groundwaters. Precipitation δ18O were obtained from estimates for
the site location made using the OIPC64). River δ18O data includes measurements
from the Heiderbach, a small stream in the Rinne valley65, the Bode, a Saale
tributary in northern Thuringia66 and the Elbe close to the confluence of the
Saale67. Spring water δ18O data includes measurements from four small springs in
the Rinne valley65. Error bars represent the overall uncertainty introduced by the
conversion to drinking water oxygen isotope values (see Supplementary Text 5).
Ndatapoints for each error bar varies by season and time bin and can be found in detail
in Supplementary Table 6. In the time bins δ18Odw estimates are based on a variable
number of tooth specimens with n36-39 ka = 7, n42-43 ka = 3, n43-45 ka = 2, n45-48 ka = 3.
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Corresponding author(s): Sarah Pederzani
Last updated by author(s): 17.12.2023
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Data collection During IRMS data collection Isodat 3.0 was used.
Data analysis Data analysis was conducted using R version 4.2.0. All analysis code is available at https://osf.io/wunfd/
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Study description New oxygen, carbon, nitrogen, zinc and strontium stable isotope data and radiocarbon dates of 16 sequentially sampled equid teeth;
zinc and strontium stable isotope data of 24 teeth from various omnivore, herbivore and carnivore taxa to reconstruct climate and
environments faced by H. sapiens groups during the Middle to Upper Palaeolithic transition at Ilsenhöhle in Ranis, Germany.
Research sample Equid teeth were targeted for serial sampling as they have high-crowned teeth and are obligate drinkers that reflect oxygen isotopes
of meteoric water. Additional teeth for a variety of carnivore, omnivore, and herbivore taxa were chosen for Zn and Sr analysis to
explore feeding ecology across large mammals in the food web. Teeth were chosen to cover the lower part of the stratigraphic
sequence, representing the MP/UP transition.
Sampling strategy For equids, a sample size of >4 teeth per archaeological unit was used to guide sampling, as this has been shown to yield sufficiently
precise palaeotemperature estimates (uncertainty of ~ 2-4 °C) from oxygen stable isotope measurements in European Palaeolithic
palimpsest contexts (see Pryor et al., 2014 Palaeo3). Teeth from other taxa were chosen from a single archaeological unit, Layer IX, as
this layer offers that largest faunal collection in the lower stratigraphic sequence. Sample sizes were constrained by availability, with
an aim of 4-5 teeth per taxon and > 8 teeth per dietary group (carnivore, omnivore, herbivore).
Data collection S. Pederzani collected equid tooth samples and conducted serial sampling, sample preparation and IRMS measurements for oxygen
stable isotope analysis. H. Fewlass and S. Talamo conducted collagen extraction, carbon and nitrogen stable isotope analysis and
radiocarbon dating on equid dentine and mandible bone samples. M. Trost collected tooth enamel samples from non-equid taxa and
conducted sample preparation for zinc and strontium stable isotope analysis. N. Bourgon and J. McCormack conducted zinc and
strontium isotope measurements.
Timing and spatial scale Two equid tooth samples from the 2016-2022 excavation were recovered in 2019 and obtained in 2020 from the Thüringer
Landesamt für Denkmalpflege und Archäologie, Weimar, Germany. No other suitable equid teeth were recovered from the 2021 or
2022 campaigns of these renewed excavations. All other tooth samples were obtained in 2018-2019 from the collection of the
1932-1938 excavation campaign housed at the Museum für Vorgeschichte, Halle (Saale), Germany. Teeth originate from a range of
squares across the extent of the excavations and square and depth information is given in Supplementary Table 1.
Data exclusions In few cases, individual oxygen stable isotope measurements (of triplicate analyses conducted for each sample) were excluded if
predetermined IRMS quality control criteria of peak shape and the relationship of sample amount to peak area did not conform to
good quality measurements. In these cases, oxygen stable isotope delta values represent the average of two, rather than the typical
three measurements per sample.
Reproducibility All stable isotope analyses (O, C, N, Zn, Sr) were repeated on at least a subset of samples (all samples in triplicate for oxygen, all
samples in duplicate for carbon and nitrogen, a subset in duplicate for Zn and Sr isotope analysis) to determine analytical
reproducibility. Details of analytical reproducibility are described in Supplementary Text 5 (Extended methods). Additionally, all code
and data to reproduce the manuscript text, figures, tables, statistical analyses, inverse modelling and temperature estimation are
supplied in an associated online repository.
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Randomization N/A
Blinding N/A
Did the study involve field work? Yes No
Field work, collection and transport
Field conditions Excavations at Ilsenhöhle in Ranis were conducted from 2016-2022. Two equid teeth used in this study were recovered in July/
August 2019.
Location All specimens were recovered from Ilsenhöhle in Ranis, Germany (50°39.7563’N, 11°33.9139’E).
Access & import/export Samples were obtained from the Thüringer Landesamt für Denkmalpflege und Archäologie (TLDA), Weimar, Germany and the
Landesamt für Denkmalpflege und Archäologie Sachsen-Anhalt, Museum für Vorgeschichte (LDA), Halle (Saale), Germany. Sampling
was conducted at the MPI-EVA, Leipzig, Germany without need for exporting. Permissions for destructive sampling were given by the
LDA on 18.04.2018 (Nr. 14/2018) and by the TLDA on 26.11.2019 (Vorgangsnummer 16/116).
Disturbance The samples were obtained from excavations of the archaeological site. The area of the renewed excavations was kept as small as
possible to reach the lowest layers following safety measures of stepped excavation levels.
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Methods
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ChIP-seq
Flow cytometry
MRI-based neuroimaging
Palaeontology and Archaeology
Specimen provenance Samples were obtained from the Thüringer Landesamt für Denkmalpflege und Archäologie (TLDA), Weimar, Germany and the
Landesamt für Denkmalpflege und Archäologie Sachsen-Anhalt, Museum für Vorgeschichte (LDA), Halle (Saale), Germany. Sampling
was conducted at the MPI-EVA, Leipzig, Germany without need for exporting. Permissions for destructive sampling were given by the
LDA on 18.04.2018 (Nr. 14/2018) and by the TLDA on 26.11.2019 (Vorgangsnummer 16/116). Specimen IDs issued by the museums
are reported for all specimens in Supplementary Table 1
Specimen deposition All specimens have been returned to the LDA and the TLDA, where they are curated under museum authority.
Dating methods 16 equid samples collected for this study were pretreated and measured for 14C dating as part of this study. Collagen extraction and
purification (including ultrafiltration) was carried out at the MPI-EVA, Leipzig using published protocols, which are described in the
methods section and the Supplementary Extended methods. The suitability of collagen extracts for
measurement was assessed based on coll % yield, elemental data (C%, N%, C:N). Quality criteria for all samples
is included in the Supplementary Extended methods and in Supplementary Table 2. Samples were graphitised and measured with a
MICADAS AMS at ETH-ZURICH and MAMS. Both uncalibrated and calibrated dates and laboratory codes are reported in
Supplementary Table 2. Dates were calibrated using OxCal 4.3 using the IntCal20 data set.
Tick this box to confirm that the raw and calibrated dates are available in the paper or in Supplementary Information.
Ethics oversight Permissions for destructive sampling were given by the LDA by the TLDA, who are the relevant archaeological authorities regulating
protection of archaeological finds in Thuringia and Saxony-Anhalt, Germany.
Note that full information on the approval of the study protocol must also be provided in the manuscript.
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