The 200,000 year long record of stable isotopes (δ18O, δ13C) of cave bear (Ursus spelaeus) teeth from Biśnik Cave, Poland

Quaternary International xxx (2013) 1e12

Contents lists available at ScienceDirect

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

The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland Maciej T. Krajcarz*, Magdalena Krajcarz Institute of Geological Sciences, Polish Academy of Sciences, Research Centre in Warsaw, Twarda St. No 51/55, PL-00818 Warszawa, Poland

a r t i c l e i n f o

a b s t r a c t

Article history: Available online xxx

The excavations in Bi
snik Cave created an opportunity to investigate changes in d13C and d18O values of Ursus spelaeus teeth from one site along a 200,000 year period of sedimentation, for the first time in Eastern Europe. Bi
snik Cave (KrakóweCze˛ stochowa Upland, southern Poland) is a multilayered archaeological and paleontological site with late Middle Pleistocene and Late Pleistocene sediments, where several important changes of Quaternary climate have been recorded. The project considered if and how detailed the isotopic data from one multilayered site record these great climatic changes. The method used was an isotopic analysis of carbonate in bioapatite from tooth enamel. Teeth of cave bear (U. spelaeus) were chosen as research material. The results showed that the d18O values of cave bear teeth from Bi
snik Cave range from
14.9 to
4.4& VPDB with some variation between particular layers. Values of d13C vary between
20.5 and
14.5& VPDB. The analysis revealed that isotopic record is diverse between different types of teeth, probably due to differences in time of growth and different impact of nursing and hibernation. The results verify the responsiveness of this species to great climatic changes during Middle and Late Pleistocene in Eastern Europe. Cave bear was an ecologically inflexible species, associated with the same type of food during 200,000 years and not able to cope with the coldest phases of the Pleistocene. Ó 2013 Elsevier Ltd and INQUA. All rights reserved.

1. Introduction

environmental factors, so the studies on isotope ratio of fossil animal tissues allow reconstruction of palaeoenvironment. The medium carrying oxygen and carbon isotopic information in fossil animal remains is biogenic apatite included in teeth enamel, and also in dentine and bones, and collagen contained in bones and dentine. Enamel, a very hard material synthesized in a relatively short time during animals life and most resistant to diagenesis among all animal tissues, is a good source of isotopic data (Quade et al., 1992; Reinhard, 1996; Koch et al., 1997). Among Middle and Upper Pleistocene terrestrial mammals, the cave bear (Ursus spelaeus Rosenmüller 1794) is one of the most especially suited species for palaeoenvironmental reconstruction based on isotopic data (Reinhard, 1996), due to the following features:

The usefulness of stable isotope analysis for reconstruction of palaeoclimate, palaeoenvironment and dietary habits of fossil animals has been confirmed by number of studies proceed during last four decades (DeNiro and Epstein, 1978; Ambrose and DeNiro, 1986; Koch et al., 1989; Bocherens et al., 1994, 1995, 1996, 2011; Hilderbrand et al., 1996; Reinhard, 1996; Stiner et al., 1998; Vila Taboada et al., 2001; Drucker et al., 2003, 2009, 2011; Richards et al., 2008; Chritz et al., 2009; García García et al., 2009; Pushkina et al., 2010; Dotsika et al., 2011; Skrzypek et al., 2011). Years of research on carbon and oxygen isotopes in animal tissues have established the model of fractionation between isotope ratios in water and food consumed by animals and isotopes that are built in the animal tissues (Bryant and Froelich, 1995; Cerling and Harris, 1999; Lee-Thorp and Sponheimer, 2003; Passey et al., 2005). The isotope composition of water and plants is controlled by

* Corresponding author. E-mail addresses: [email protected] twarda.pan.pl (M. Krajcarz).

(M.T.

Krajcarz),

magdakraj@

- the remains of cave bear are the most numerous Pleistocene fossil remains in Europe (Bocherens et al., 2011); - it was a widespread species, with geographic distribution ranging from Iberian Peninsula to Ural Mountains and from Mediterranean Sea to Baltic Sea (Kurtén, 1968; Kahlke, 1999); - it was an eurytopic species, able to live in different environmental conditions e from glacial to interglacial and from forests to steppe-tundra (Kurtén, 1968; Reinhard, 1996);

1040-6182/$ e see front matter Ó 2013 Elsevier Ltd and INQUA. All rights reserved. http://dx.doi.org/10.1016/j.quaint.2013.07.022

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

2

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

- cave bear teeth are large enough to provide samples for isotopic analysis (Reinhard, 1996). The authors embarked on the following project to determine if cave bear teeth from one multilayered site really record the climatic changes, and in what detail? The goal was to verify if isotopic biochemistry of the species was responsive or resistant to great climatic turnovers during Middle and Late Pleistocene. To achieve such a goal, a special site was required e multilayered cave sediments with a continuous sequence of deposits that originated during a long period of the Pleistocene, and with numerous cave bear remains in each layer. The authors chose Bi
snik Cave, a site that fulfills these requirements. 2. Material and methods 2.1. The site Bi
snik Cave is a multilayered archaeological site situated in southern Poland (Fig. 1). The deposits are built of numerous layers dated to Neogene/Early Pleistocene, Middle Pleistocene, Late Pleistocene, and Holocene (Fig. 2). Due to occurrence of over a dozen Middle Palaeolithic levels, the site is highly important for archaeology of Neanderthals (Cyrek et al., 2010). The cave has been excavated since 1991 and yielded hundreds of stone tools, several fire places, and over one hundred thousand animal bones. Several important changes of Pleistocene climate are recorded in Bi
snik Cave. Aeolian loess or loess-like sediments (layers Nos. 2, 12, 16, intercalations inside layer No. 15), with traces of solifluction processes, represent severe or even periglacial environments (Miros1aw-Grabowska, 2002; Krajcarz and Madeyska, 2012).

Permafrost melting followed by water inflow (Miros1awGrabowska, 2002) or mud flows (Krajcarz and Cyrek, 2011) are recorded (layers Nos. D, E, S1eS2, 5, 6, 7, 19d and 19bc). Other layers indicate warm climatic conditions (especially layers Nos. 9, 11, 13a, 14, 19, 19a), when dense forests grew around the cave (Cyrek et al., 2010; Krajcarz et al., 2010). Other layers are related to average climatic conditions. Large climatic and environmental turnovers around Bi
snik Cave were confirmed by diverse geologicalpaleontological analyses, including: lithological features, limestone rubble weathering state, bone weathering state, clay mineral ratios, secondary precipitations in sediments, molecular fossils, and paleoecology of micromammals and large mammals (Miros1awGrabowska, 2002; Wiszniowska et al., 2002; Krajcarz, 2009; Cyrek et al., 2010; Krajcarz et al., 2010; Krajcarz and Cyrek, 2011; Krajcarz and Madeyska, 2012). The age of sediments was established on the basis of multiple methods: climatostratigraphy, biostratigraphy, uranium/thorium series dating, radiocarbon dating, and thermoluminescence dating (Hercman and Gorka, 2002; Miros1aw-Grabowska, 2002; Cyrek et al., 2010; Krajcarz and Madeyska, 2012). Although the layers from Bi
snik Cave are related to diversified environments, almost all yielded numerous cave bear remains. U. spelaeus is the most abundant species among the Pleistocene fossils from Bi
snik Cave, as its remains consist of almost 90% of identifiable bones of large mammals (Marciszak et al., 2011; Marciszak, 2012). 2.2. Landscape and palaeolandscape around Bi
snik Cave The palaeogeography of the nearby landscape is really important when trying to interpret the d18O variations seen in different

Fig. 1. Localization of the Bi
snik Cave: A e current surrounding topography; B e localization on a map of Poland; C e the closest occurrence of ice sheet and glacier-derived waters _ during last 300,000 years (according to Lamparski, 1961; Rózycki, 1982; Lewandowski, 1987, 2011; Mojski, 2005; age interpretation according to Lindner and Marks, 2012).

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

3

Fig. 2. Plan of the Bi
snik Cave and profile of sediments. Stratigraphy according to Krajcarz and Madeyska (2012) and Krajcarz and Cyrek (2011). Bear icons indicate layers where cave bear teeth occur and were taken to isotopic analysis.

layers of a researched site. The possible change in sources of water could be a more important factor of d18O differentiation than temperature change if the site is located in a mountainous area or other area where glaciers and glacier-derived water were occasionally present (see for example the case of the Totes Gebirge in Austria, Bocherens et al., 2011). Glacier ice is depleted in 18O (Michener and Lajtha, 2007) and therefore melting water exhibits low d18O values. If such water is ingested by an animal, the isotopic signal recorded in its tissues is modified and not longer relates to air temperature or the isotopic signal of meteoric water. Bi
snik Cave is situated in a diversified landscape of the Polish Jura, with numerous limestone monadnock hills separated by deep valleys. Average relief between valley bottoms and neighboring monadnock tops is about 50 m. Altitudes in the area vary from about 230 m a.s.l. at bottoms of the largest rivers valleys, to 515.5 m a.s.l. at the highest culmination of the Góra Zamkowa hill (Fig. 1: A). The cave is situated at 410 m a.s.l. These relatively low altitudes did not induce the development of mountain glaciers during the Pleistocene. In addition, the Polish Jura was never covered by an ice sheet during the Pleistocene but was emerged as a concave nunatak (Lewandowski, 2011). The closest ice sheet during the time interval of Pleistocene recorded in Bi
snik Cave occurred during Odranian stadial, previously dated to MIS 8 (Lindner et al., 1995; Mojski, 2005) but in the newest stratigraphic scheme of the Quaternary of Poland (Lindner and Marks, 2012) correlated with the early part of MIS 6. The maximum range of the Odranian ice sheet was

situated about 40 km to the west and north from Bi
snik Cave, and fluvioglacial drainage flowed about 25 km away (Fig. 1: C). This episode may be correlated with deposition of layer No. 18 in the Bi
snik Cave (Krajcarz and Madeyska, 2012). During later cold episodes (i.e. Warthanian stadial, late part of MIS 6, and Vistulian Glaciation, MIS 4 and 2) the ice sheet ranges and routes of melting water were situated far to the north (Mojski, 2005; Lindner and Marks, 2012). The location of Bi
snik Cave suggests that bears never used the glacier-derived water as a source of drinking water during the last 250,000 years. 2.3. Material Bones from Bi
snik Cave are not suitable for isotopic analysis. The C/N ratio for over half of the bones is 4.0e18.0. This indicates strongly weathered material (see DeNiro, 1985; Bocherens et al., 1995). For this reason, only teeth enamel was analyzed. For this study, 121 samples of teeth from adult cave bears were chosen (Table 1). All teeth came from the NE part of Side Chamber and S part of Main Chamber (Fig. 2). Only remains from layers that do not occur in these parts of the cave (layer No. 4) were chosen from the ‘At Overhang’ place. Though the number of cave bear teeth found at the site is quite large, it differs strongly between layers. For this reason, it was impossible to select the same number and the same type of teeth from each layer. That is why variation in isotopic composition of teeth arising during time of life must be taken in

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

4

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

consideration while interpreting the results. The teeth were taken by authors in field work during archaeological excavations in 2010e 2012. Teeth were identified as U. spelaeus by their morphology (Altuna, 1973; Torres et al., 1978; Torres, 1988). However, the DNA analyses of cave bear from Bi
snik Cave has not been conducted, so it is possible that the material may represent several closely related taxa of species or subspecies rank, such as Ursus ingressus, U. spelaeus spelaeus, U. spelaeus ladinicus or U. spelaeus eremus (Marciszak et al., 2011).

Cave bear teeth and bones were found in layers from No. 19bc to No. 1. However, some of the youngest layers (Nos. S1eS2, 5e6, 7) were accumulated by fluvial and colluvial processes (Miros1awGrabowska, 2002; Krajcarz and Madeyska, 2012), so the bear remains in these layers may be redeposited and not represent the time of layer accumulation. The oldest Middle Pleistocene layers Nos. 19bc and 19d are built of mud flow sediments (Krajcarz and Cyrek, 2011). The teeth of U. spelaeus are mixed in these sediments with remains of Ursus deningeri and other fauna from early

Table 1 List of teeth samples from Bi
snik Cave and results of isotopic analysis. Sample

Inv. number

Ursus spelaeus IB-170 W-2442 IB-17 W-9802 IB-19 W-9824 IB-20 W-9894 IB-21 W-9895 IB-22 W-9941 IB-196 ING IB-196 IB-23 W-10030 IB-24 W-10082 IB-25 W-10138 IB-101 W-10088 IB-102 W-10164 IB-103 W-8525 IB-111 W-8816 IB-26 W-9464 IB-27 W-9469 IB-28 W-9573 IB-74 W-9226 IB-75 W-9194 IB-105 W-9055 IB-106 W-9232 IB-107 W-9229 IB-108 W-9056 IB-109 W-9261 IB-110 W-9257 IB-29 W-9590 IB-30 W-9761 IB-31 W-9762 IB-32 W-9764 IB-121 ING IB-121 IB-122 W-9417 IB-124 ING IB-124 IB-125 ING IB-125 IB-126 ING IB-126 IB-188 ING IB-125 IB-33 W-9440 IB-127 W-9477 IB-130 M-171/11 IB-131 W-9470 IB-132 W-10190 IB-34 W-9439 IB-35 W-9458 IB-36 W-9486 IB-37 W-9497 IB-38 W-9511 IB-39 W-9517 IB-40 W-9520 IB-41 W-9560 IB-42 W-9637 IB-43 W-9676 IB-190 ING IB-190 IB-45 W-9774 IB-182 W-9550 IB-192 ING IB-192 IB-48 W-9602 IB-49 W-9720 IB-51 W-9872 IB-52 W-9878 IB-53 W-9924 IB-54 W-9770 IB-55 W-9897

Layer

Tooth

d13CC (&, VPDB)

d18OC (&, VPDB)

Sample

Inv. number

Layer

Tooth

d13CC (&, VPDB)

d18OC (&, VPDB)

4 S1eS2 S1eS2 S1eS2 S1eS2 S1eS2 5e6 5e6 5e6 7 7 7 7 7 9 9 9 9 9 9 9 9 9 9 9 10 10 10 10 10 10 10 10 10 10 10a 10a 10a 10a 10a 11 11 11 11 11 11 11 11 11 11 11 12 12 12 12 12 12 12 12 12 12

i3 m2 m2 c m2 m2 I2 I3 m2 M2 C I2 I3 i3 I2 i3 i3 i1 M2 i1 i3 i3 i2 i3 I2 m2 m2 m2 m2 I2 m1 i2 m3 m1 m3 i2 i2 C m2 I3 M2 M2 I3 i3 m1 i2 i3 I3 m2 m2 P4 M1 m1 m3 M1 M1 M2 I1 m1 m2 I2


16.20
17.89
17.17
18.27
18.93
16.39
14.46
15.59
15.62
17.22
17.88
17.34
17.51
16.52
16.29
15.98
16.63
17.52
16.78
16.43
18.15
16.28
16.46
17.14
16.90
19.79
19.76
19.04
17.24
15.73
18.08
17.14
17.16
18.18
17.34
17.25
16.28
18.14
15.60
18.49
17.02
17.62
20.27
19.89
17.96
15.9
15.39
18.55
16.2
17.2
17.55
16.74
18.40
16.02
17.6
17.7
17.3
17.73
20.24
16.63
17.05


7.00
11.07
11.85
8.60
12.14
9.09
6.76
10.02
7.63
10.90
8.22
6.86
7.07
6.58
6.43
6.70
7.16
7.47
5.7
6.52
6.17
6.66
6.16
7.53
6.24
11.44
14.57
10.34
6.89
6.29
6.73
6.20
5.28
7.60
5.59
9.78
6.14
6.29
4.41
6.03
7.43
8.28
14.93
12.20
8.97
5.82
7.05
6.37
5.62
6.27
4.58
6.76
8.04
9.21
6.47
6.62
8.48
6.54
6.21
7.20
6.98

IB-56 IB-57 IB-76 IB-77 IB-78 IB-79 IB-136 IB-137 IB-138 IB-139 IB-171 IB-58 IB-82 IB-113 IB-114 IB-141 IB-142 IB-143 IB-172 IB-173 IB-183 IB-174 IB-175 IB-176 IB-177 IB-178 IB-179 IB-181 IB-189 IB-90 IB-91 IB-92 IB-116 IB-93 IB-96 IB-97 IB-59 IB-60 IB-61 IB-62 IB-63 IB-64 IB-65 IB-98 IB-147 IB-148 IB-149 IB-184 IB-187 IB-193 IB-66 IB-67 IB-68 IB-69 IB-70 IB-71 IB-152 IB-118 IB-72 IB-73

W-9860 W-10147 W-8784 W-10298 W-10397 W-8777 W-10150 W-10027 M-810/11 W-10049 M-831/11 W-10032 W-8644 W-8760 W-8638 W-10314 W-10317 W-10321 ING IB-172 ING IB-173 W-10057 B96/97/1 B/49/99/1 B/50/99 B/92/97/36 B96/97/5 B7/27/25 B5/97/29 B96/97/5 ING IB-90 W-9683 W-9125 W-9060 ING IB-93 ING IB-96 W-9737 W-9793 W-9796 W-9837 W-9912 W-9915 W-9929 W-9932 ING IB-98 ING IB-147 ING IB-148 ING IB-149 W-9931 ING IB-149 ING IB-193 W-10017 W-10044 W-9946 W-9967 W-9975 W-9982 ING IB-152 W-8677 W-10077 W-10128

12 13 13 13 13 13 13 13 13 13 13 13a 13a 13a 13a 13a 13a 13a 13a 13a 13a 14 14 14 14 14 14 14 14 15 15 15 15 16 18 18 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19a 19a 19a 19a 19a 19a 19a 19bc 19bc 19bc

m1 i3 m1 m2 m1 I2 C m1 M m2 m3 P4 p4 p4 i3 C i3 C i2 i2 C i3 i3 i1 i3 I2 I2 I2 I2 I3 I2 m1 I2 m1 m2 m3 m2 m2 m1 m1 m1 m3 I2 C C m1 C m2 C I1 M2 M1 C i1 I3 M2 C i2 I1 m3


18.18
19.39
16.75
16.74
19.75
15.87
18.58
18.80
16.99
17.68
16.94
19.5
19.05
15.83
20.45
19.21
15.70
17.71
18.08
16.32
17.78
16.27
16.85
16.62
15.67
17.91
16.12
16.08
17.86
17.08
17.52
17.12
16.47
18.28
17.06
18.17
17.31
16.46
17.86
15.91
17.07
19.1
16.76
18.46
17.33
16.60
18.61
16.87
18.52
15.57
16.64
19.02
18.18
17.72
18.35
17.15
17.68
16.74
16.27
18.50


7.79
6.04
5.48
6.95
6.38
7.74
6.89
5.44
6.83
8.18
5.84
5.94
8.11
7.32
7.14
7.50
6.35
6.65
7.14
6.95
6.80
7.74
6.99
5.02
8.62
7.07
7.79
6.48
6.79
6.89
7.06
6.93
7.16
6.04
5.31
6.57
7.75
7.72
7.01
5.10
5.84
9.13
5.33
6.73
5.03
5.49
5.78
5.40
5.65
4.69
5.83
7.18
5.03
5.77
8.10
5.59
5.27
5.38
6.11
5.68

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

5

Table 1 (continued ) Sample

Inv. number

Ursus arctos IB-099 W-9432 IB-161 ING IB-161 Rangifer tarandus IB-162 B/186/96/12 IB-167 B/159/12 IB-104 W-8525

Layer

Tooth

d13CC (&, VPDB)

d18OC (&, VPDB)

1 15/16

m2 m1


16.7
16.7


6.4
5.3

2 2/4 19bc

m2 m3 P2


8.4
9.6
8.1


5.3
9.6
7.5

Middle Pleistocene, including Panthera fossilis (Marciszak and Stefaniak, 2010) and Panthera onca gombaszoegensis (Marciszak et al., 2011). Remains from these layers may be redeposited and older than the layer itself. The youngest layers (Nos. 1, 2, 3, A, B, C, D, E) are dated to MIS 2 and MIS 1, so they originated after cave bear extinction (see Pacher and Stuart, 2009). The single cave bear remains known from these layers (Wiszniowska et al., 2002; Marciszak et al., 2011) are undoubtedly redeposited by secondary processes and were not analyzed. 2.4. Enamel preparation and isotopic measurements The method used was an isotopic analysis of carbonate in bioapatite (Ca5(PO4,CO3)3O,OH) from teeth enamel. The enamel was chosen as the source because it is believed to be more

Sample

Inv. number

Layer

Tooth

d13CC (&, VPDB)

d18OC (&, VPDB)

resistant to post sedimentary diagenetic alteration than dentine or bones (Quade et al., 1992; Reinhard, 1996; Koch et al., 1997; Stiner et al., 1998). Enamel from teeth was collected by drilling about 10 mg powdered samples. The samples were analyzed according to the method described by Koch et al. (1997) and Bocherens et al. (1996). First, the enamel was soaked in 3% NaOCl solution for 24 h in room temperature to remove organic contamination. Then, it was rinsed with distilled water and treated with 1 M acetic acid for 72 h to eliminate exogenous carbonate. Finally, samples were rinsed with distilled water and dried. Carbon dioxide was extracted by dissolution in 100% H3PO3 in KIEL IV device and measured on Finnigan Delta þ mass spectrometer at the Isotope Dating and Environment Research Laboratory, Institute of Geological Sciences, Polish Academy of Sciences, in Warsaw (Poland).

Fig. 3. d18O and d13C values achieved for particular layers from Bi
snik Cave and anomaly of layers’ mean values in respect to the mean of the whole profile. Mean is
7.1& for d18O and
17.4& for d13C.

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

6

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

Fig. 4. d18O values of enamel from different types of cave bear teeth achieved for particular layers from Bi
snik Cave.

The isotopic ratios are expressed as d13C and d18O according to: d X ¼ (Rsample/Rstandard  1)  1000, where X is an element (C or O), A is nucleon number and R is isotopic ratio 13C/12C or 18O/16O. Values are reported in relation to international standard VPDB. Used laboratory standards were NBS19, NBS18 and LSVEC. Isotopic results for different anatomical tooth types were compared statistically using the non-parametric ManneWhitney U-test (PAST software), which is a test used in isotopic studies (Bocherens et al., 2011). A

3. Results 3.1. Reproducibility of analytical method Replicate analyses of enamel were performed on three teeth (Table 2) to control the reproducibility of applied preparation and measurement methods. For all replicated teeth, the d13C values differ by less than 0.2&. Differences in d18O values are about 0.3& for two teeth and less than 0.2& for one tooth. These differences show good reproducibility of preparations and measurements, and equal the reproducibility obtained in other studies (Reinhard, 1996; Bocherens et al., 2011). Table 2 d13C and d18O values yield for teeth on which replicate preparations and measurements were done Teeth

Sample d13C

Difference in d13C value (&)

d18O Difference in d18O value (&)

IB-125
17.16 0.18


5.28 0.31

IB-188
17.34 IB-149
18.61 0.09


5.59
5.78 0.13

IB-187
18.52 B96/97/ IB-178
17.91 0.05 5 IB-189
17.86


5.65
7.07 0.28

ING IB125 ING IB149


6.79

is
0.31. Ranges for particular layers differ in maximal, minimal, and mean values (Fig. 3), especially distinctly in minimal values. The lowest minimal and mean values are observed for layers Nos. S1eS2, 5e6, 7 and 10. Differences in maximal values are less clearly marked. Differences between layers are more distinct in the upper part of profile, over the layer No. 13. Deviations of layers’ mean d18O values from the mean of whole profile are distinct and differentiated (Fig. 3). This demonstrates that the primary isotopic signal, different between climatostratigraphic units, was not covered by bear physiology during life or by later diagenetic processes. The lower part of profile including layer No. 13 along with all lower layers expresses homogenization of d18O values. The curve of layers’ means deviation from the mean of the entire profile, although it shows some trend, is smoothed in the lower part of profile. The analyzed samples represent all type of cave bear teeth, except small premolars (p1ep3). The d18O values obtained for first incisors ranged from
7.5 to
4.7&, and for second incisors between
9.8 and
5.3&. Both types of teeth showed no clear differences in isotopic composition of oxygen between layers (Fig. 4, Table 3). The same situation may be observed with reference to canines and first and third molars. The d18O values for canines range from
8.0 to
5.0&, for first molar from
9.0 to
5.1&, and for third molar from
9.2 to
5.3&. Table 3 Statistical comparisons of d18O data obtained for different anatomical positions of analyzed teeth (p values by ManneWhitney test). Tooth type i2 or I2 i3 or I3 c or C

p4 or P4 m1 or M1 m2 or M2 m3

i1 or I1 i2 or I2 i3 or I3 c or C p4 or P4 m1 or M1 m2 or M2

0.7768 0.9716 0.4550 0.9548 e e e

0.1080 e e e e e e

0.0116 0.1022 e e e e e

0.3413 0.3927 0.0534 e e e e

0.1836 0.6849 0.0578 0.6868 0.9676 e e

0.0364 0.0595 0.5833 0.0411 0.2855 0.0468 e

0.6093 0.3717 0.0923 0.9683 0.9247 0.7724 0.2255

3.2. Oxygen isotopic values The d18O values of the carbonate fraction (d18OC) ranged from
14.9 to
4.4& VPDB (Table 1). Coefficient of variation

A different situation is observed for the third incisor and second molar. The ranges of d18O values are respectively
19.2 to
6.0& and
14.6 to
4.4&. The ranges are much wider than for other

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

teeth and vary distinctly between layers (Fig. 4). These teeth do not exhibit any statistically significant differences between them in d18O, but show differences when compared with other tooth types (Table 3). Both these teeth show the most negative values for layer No. 10. The number of samples from fourth premolars is too small to assess their variability. 3.3. Carbon isotopic values The d13C values of carbonate fraction (d13CC) range from
20.5 to
14.5& VPDB (Fig. 3, Table 1). Coefficient of variation is
0.14. No systematic pattern can be observed in the isotopic signature for layers. The d13C record follows systematic changes between particular types of teeth shown by oxygen (Fig. 5). The d13C values obtained for first incisors ranged from
17.73 to
15.57& and for second incisors between
18.08 and
14.46&. The range for canines is
19.21& to 17.33& and for first molars
20.24 to
15.91. Third incisors and second molars give the widest ranges of d13C values (
20.45 to
15.39& and
19.79 to
15.6&). No statistically significant difference can be found between d13C values of third incisors and second molars; however, differences between these teeth and the other ones are distinct (Table 4).

Table 4 Statistical comparisons of d13C data obtained for different anatomical positions of analyzed teeth (p values by ManneWhitney test). Tooth type i2 or I2 i3 or I3 c or C

p4 or P4 m1 or M1 m2 or M2 m3

i1 or I1 i2 or I2 i3 or I3 c or C p4 or P4 m1 or M1 m2 or M2

0.2986 0.1452 0.4555 0.9549 e e e

0.7597 e e e e e e

0.6103 0.3072 e e e e e

0.0026 0.0001 0.0538 e e e e

0.2074 0.0004 0.1699 0.3000 0.9031 e e

0.4409 0.0671 0.7019 0.0009 0.3141 0.0456 e

0.2502 0.0415 0.4599 0.1779 0.6366 0.7287 0.3667

4. Discussion 4.1. Isotopic record from anatomically different teeth The dental eruption sequence for cave bear is unknown. It is assumed that milk teeth replacement in cave bear was similar to modern brown bear, in which case the dental eruption is well recognize (Andrews and Turner, 1992). This assumption is possible

7

as these two species are relatively closely related, derived from one ancestor (Kurtén, 1968). For isotopic analysis, apart from time of teeth eruption, the time of enamel formation (maturation) is very important. Most enamel maturation is carried out before eruption of teeth, and may continue slowly during eruption and after (Hillson, 2005). Brown bear teeth erupt in different months and with different speeds. Studies on other species showed that the time of enamel formation is also different for different teeth (Fricke and O’Neil, 1996). The enamel during its development records conditions of different seasons and different weather. In case of brown bears and cave bears, there are two physiological factors that may have important impact on isotopic record of teeth: nursing and dormancy (Bocherens et al., 1994; Nelson et al., 1998; FernándezMosquera et al., 2001). Enamel of some teeth forms when bears are still relying on mother’s milk. These teeth may have different isotopic records than those erupting later. Furthermore, the enamel may be formed partly during winter hibernation. During this period the animal does not take food or water, its metabolism is modified, and isotopic composition of growing tissues changed regardless of environmental conditions (Bocherens, 2004). Impact of nursing and hibernation on a d13C record is a well known issue (Wright and Schwarcz, 1999; Bocherens, 2004; PérezRama et al., 2011). Both nursing and dormancy cause depletion in 13 C, and the difference between cubs and adult bears is 2.2&. The oxygen isotopes are fractionated in animal body, as lighter 16 O is lost due to respiration and body tissues are enriched in 18O (Wright and Schwarcz, 1999). The main source of oxygen for animals is ingested water (Bryant and Froelich, 1995). When an animal is active and imbibes external water, the relation between d18O values in animal body and in environment are stabilized. However, during hibernation the animal does not assimilate water from the environment and metabolizes only the water accumulated in a body during the warm season. Oxygen fractionation should therefore increase d18O values in bears during hibernation, as well as in other hibernating species. The exact relationship between time of enamel formation and tooth eruption for modern bears was established only for canines (Rausch, 1969). Times for other types of teeth may be only hypothetically estimated on the basis of notices given by Rausch (1969) and time of eruption (Fig. 6). The hypothetic time of enamel formation revealed that each permanent tooth of a bear is affected by both nursing and mother’s hibernation during enamel maturation. As Fricke and O’Neil (1996) assumed, the mineralization interval of each tooth is limited by the time of eruption. In the case of teeth that erupt first, the timespan of enamel formation may fall

Fig. 5. d13C values of enamel from different types of cave bear teeth achieved for particular layers from Bi
snik Cave. Symbols are the same as for Fig. 4.

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

8

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

Fig. 6. Estimated time of cave bear tooth formation with hypothetic changes of d18O signal in tooth enamel during bear ontogenesis.

completely during the cold period, when mothers’ metabolism and milk are affected by hibernation. This problem may concern first molars (m1/M1) and first and second incisors (i1/I1, i2/I2) as these teeth are formed earliest during bear ontogenesis. They start to erupt when cubs are about 5e6 months old, a time when mothers with cubs usually abandon their dens. Teeth that erupt several months after the end of mother’s hibernation also record an impact of hibernation, as was reported by Rausch (1969) on example of canines. However, the spring and summer seasons when the female drinks environmental water and eats regular food may also be recorded. Fourth premolars (p4/P4) start to erupt about the 7th month, and third incisors (i3/I3) and second molars (m2/M2) about the 8th month. The time delay between growing of these teeth and growing of i1/I1, i2/I2 and m1/M1 is not long; however it lasts for several months. Third molars (m3) and canines (c/C) erupt as the last ones. The time of eruption of these teeth falls in either the warm season or the second winter of the cubs. Third incisors (i3/I3), second molars (m2/M2), fourth premolars (p4/P4) and especially third molars (m3) and canines (c/C) should record changes in mother’s behavior and should demonstrate the widest range of isotopic values. In the case of Bi
snik Cave, only i1/I1, i2/I2, i3/I3, m1/M1 and m2/ M2 follow the hypothetical pattern. Among them, i1/I1, i2/I2 and m1/M1 show the narrow range of d18O (Fig. 7), likely an effect of nursing during mother’s hibernation; while i3/I3 and m2/M2 exhibit relatively wide range of d18O (Fig. 7), reflecting an impact of both nursing during mother’s hibernation and nursing during mother’s activity on enamel formation. Canines, fourth premolars and third molars do not follow the hypothetical pattern, as they show narrow range of d18O (Fig. 7) although the enamel formation occurred partially during mother’s activity season. Perhaps the number of samples was insufficient and the problem needs further study. The differences in d13C values between particular tooth types are much less visible than in d18O, with modulus of coefficient of variation for d18O exceeding the value for d13C by over two times.

Although the absolute values of d18O in bear enamel do not directly reflect the environment conditions, as they are changed by mother’s metabolism, the isotopic signal in mother body is indirectly related to the water ingested by the mother before hibernation. Therefore, an indirect correlation between isotopic signal recorded in tooth enamel and palaeoenvironment can be recognized. 4.2. Is d18O in bear teeth enamel a record of palaeoclimate? Metabolic fractionation and incorporation of the isotopes into tissues is mainly controlled by animal metabolism. However, final isotope composition of tissues is related to composition of local meteoric water, drunk or taken with food by the animal (Bryant and Froelich, 1995; Kohn et al., 1996, 1998; Wang et al., 2008; Dotsika et al., 2011). Isotopic composition of water is controlled by environmental factors, including air temperature. Ultimately, d18O in animal tooth enamel is related to the climate. This relation was widely used for palaeoenvironmental reconstruction made on the

Fig. 7. Ranges of d18O values of cave bear teeth from Bi
snik Cave, with respect to anatomical position of teeth.

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

basis of isotope composition of oxygen in fossil animal tissues (Koch et al., 1989; Bocherens et al., 1995, 1996, 2011; Reinhard, 1996; Chritz et al., 2009; Drucker et al., 2009; García García et al., 2009; Dotsika et al., 2011; Skrzypek et al., 2011). The curves of d18O changes in profile achieved for all teeth types, and for third incisors and second molars only, are similar (Fig. 8). In both cases a sharp curve is visible above layer No. 13 (about 110,000 BP), with a near straight line below that layer. From the climatostratigraphic point of view, greater climatic changes are recorded in the lower part, deposited upon glacial-interglacial turnovers (MIS 7eMIS 6eMIS 5e, Miros1aw-Grabowska, 2002; Cyrek et al., 2010; Krajcarz and Madeyska, 2012). Above layer No. 13, only intraWeichselian stadial-interstadial changes are recorded, excluding Holocene layers that do not contain cave bear remains. d18O record is reverse to the expected situation. The following hypotheses can be presented to explain this:

Fig. 8. d18O changes in the sediment profile of Bi
snik Cave. Note the sharp shape of d18O curves in the upper part of profile and straight shape in the lower part. Isotopic stratigraphy (Marine Isotope Stages) is based on different geological and paleontological data according to Cyrek et al. (2010) and modified after Krajcarz and Cyrek (2011) and Krajcarz and Madeyska (2012). Achieved d18O curves correspond to this stratigraphy especially between layers Nos. 7 and 13a.

9

1. the physiology of cave bear underwent important changes in isotope fractionation between the period of deposition of the lower and upper parts of the sediments; 2. there was/were change(s) in chronology of hibernation (time during a year when it started and time when it ended) due to climatic changes; 3. lack of ecological flexibility in cave bear, therefore cave bear remains may represent only sporadic short time intervals when environmental conditions were suitable and quite similar everytime; 4. change in water sources between cold and temperate phases; 5. after deposition of the lower part of sediments, but before the deposition of the upper part, there was a diagenetic event that homogenised d18O record in the lower layers; the isotopic record in the upper layers is not disturbed. There are no arguments to support the first hypothesis. On the basis of teeth morphology, no important evolutionary changes occurred in cave bear from Bi
snik Cave during the discussed period (Marciszak et al., 2011). Numerous finds of bear remains in cave both in the lower and upper parts of profile, along with milk teeth, suggest that there were also no important changes in general dormancy behaviour. Changes in chronology of hibernation are hard to prove for an extinct species, although possible. During colder periods, i.e. MIS 6, the warm season was shorter and hibernation longer. The prolongation of hibernation length means that enamel formation of lately forming teeth is more affected by dormancy in relation to periods with shorter time of hibernation. As a result, d18O values recorded in tooth enamel of bears (and other hibernating species) were likely higher for cold periods, even though the abiotic environments exhibited lower values, i.e. in meteoric waters. Lack of ecological flexibility for the cave bear may mean that the coldest periods are not recorded in isotopic record. It is likely that cave bear was present near Bi
snik Cave only during this time range when environmental conditions were suitable, and avoided this area during severe climatic phases. Cave bear remains are less numerous in sediments dated to the coldest stadials (layer No. 8 e MIS 4, layer Nos. 15e16e18 e MIS 6) and they may represent the short warmest intervals during these coldest stadials. On the other hand, the stadials of Early Weichselian (layer No. 10 e MIS 5b, layers Nos. 12e13 e MIS 5d) were much milder than MIS 4 and MIS 6, and bears might be present throughout these periods. The changes in water sources between cold and temperate phases may be excluded, as neither glaciers nor proglacial waters were present in the vicinity of Bi
snik Cave. The role of diagenesis in preservation of isotopic record is an issue emphasized by numerous researchers (Schoeninger and DeNiro, 1982; Nelson et al., 1986; Stiner et al., 1998; Sponheimer and Lee-Thorp, 1999). The upper part of the profile, where differences in d18O values are distinct from layer to layer, may represent not altered or only slightly altered d18O signal and are compatible with palaeoclimatic reconstruction based on other methods. Layer No. 13 is the uppermost sediment formed during Eemian Interglacial (MIS 5e), the warmest period of Quaternary history recorded in Bi
snik Cave (Cyrek et al., 2010; Krajcarz and Madeyska, 2012). The high values obtained for layers Nos. 13a and 14 may show the expected warm conditions of Eemian Interglacial (MIS 5e). Older sediments with cave bear teeth (layers Nos. 15 to 19bc) are situated below Eemian layers. This may suggest that the warm interglacial climate caused diagenetic alteration of older sediment, and the d18O record in lower layers is diagenetically modified and not related to palaeoclimate. However, previous studies demonstrated that enamel is stable enough to preserve the biogenic isotopic signal under warm climatic conditions (Bocherens et al., 1996). To

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

10

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

Fig. 9. Changes of d13C in profile of Bi
snik Cave on the background of palaeovegetation reconstructed on the basis of n-alkanes analysis (data adopted from Krajcarz et al., 2010). Changes of d13C values are slight and do not repeat a trend shown by n-alkanes CPI or n-alkanes ratio.

verify the possibility of alteration in lower layers, several teeth of other species from both the upper and lower part of the profile were sampled (Table 1). Comparison of d18O values exhibited by the reindeer (Rangifer tarandus) and the brown bear (Ursus arctos) from both lower and upper parts of the profile show similar values. The similarity of isotopic values in species between different parts of the profile argues against postburial alternation of the samples, and the possible alternation is not connected with position in the profile. The lack of variations of d18O values from bear enamel in the long section of sediments is not a unique situation to Bi
snik Cave. Similar observations were also noted by Bocherens et al. (1991) and explained as either a consequence of hibernation behaviour or an effect of diagenetic alteration. 4.3. d13C e a record of cave bear feeding behavior during 200,000 years Although cave bear represents order Carnivora, that includes many specialized predators, it is generally believed to be a herbivorous animal (Kurtén, 1968; Mattson, 1998; Bocherens et al., 2011). The obtained isotopic signature of d13C for cave bears from Bi
snik Cave does not differ from that which has been generally presented for enamel or bone collagen (Bocherens et al., 1994, 2011; Nelson et al., 1998; Fernández-Mosquera et al., 2001; Richards et al., 2008; Ábelová and Sabol, 2009) and shows a preferentially diet based on C3 plants. At Bi
snik Cave, there is no visible response in carbon isotope composition to environmental conditions that changed through the Pleistocene. There is no correlation between d13C and d18O (R2 ¼ 0.1059). Even for the upper part of the profile, the correlation is poor (R2 ¼ 0.2564). The d13C signal in cave bear, unlike d18O, was not dependent on climatic factors. This situation may be a consequence of the cave bear feeding habits. The explanation is that the food resources of cave bear

during the 200,000 year timespan were always within the spectra of C3 plants, regardless of climatic and vegetation changes. Such changes undoubtedly occurred in the vicinity of Bi
snik Cave and were partially reconstructed (Fig. 9, see also Krajcarz et al., 2010). Although the reconstruction of palaeovegetation was made mostly for the lower part of profile, the d13C values are homogenous in the whole profile. It is probable that cave bears fed constantly on the same kind of food which was available during different climatic phases. However that issue needs more research on the details of cave bear diet and isotopic composition of Pleistocene plants. 5. Conclusions This study provides the following notes on the oxygen and carbon isotopic record of cave bear teeth, and their responsiveness to climatic changes. Enamel of cave bear teeth records climatic changes in d18O values. Cave bear is a species responsive to climatic turnovers, partially affected by nursing and dormancy. In Bi
snik Cave, the d18O values from cave bear teeth excavated from the upper part of cave sediments (Weichselian, MIS 5deMIS 3) give a record of palaeoclimate compatible with sedimentological methods (i.e. Krajcarz and Madeyska, 2012). However, the climatic changes in the lower part of the profile, especially MIS 6, are not clearly recorded. This indicates that the usefulness of enamel isotopic analysis to palaeoenvironmental reconstruction may differ between different sections of the same profile in a one site. It may be due to different factors, as changes in chronology of hibernation or impact of climate on bears’ occupancy of the area. During long periods of relatively stabilized and warm climate (i.e. MIS 5) the bears might be constantly present around the cave and record long sequences of palaeoclimatic changes. However, in sediments deposited during cold phases of Pleistocene (i.e. MIS 6), the isotopic record may be discontinuous as cave bears could have been living in the cave only

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

when conditions were most suitable. The constant isotopic signal, visible as an archaeozoological record, may not represent a constant paleoenvironment, but rather short time intervals. In Bi
snik Cave, the isotopic record varies between different anatomical types of U. spelaeus teeth, due to differences in time of tooth formation. In this study, the isotopic signal of third incisors and second molars gives the widest ranges of values. The isotopic signal in bear teeth is modified by nursing and/or hibernation and may not be directly transposed to quantitative palaeoclimatic parameters. However, qualitative changes of palaeoenvironment may still be readable. The ecological flexibility of cave bear and the factors causing the diagenetic alterations of teeth need further detailed studies. The case of Bi
snik Cave has verified the formerly noticed assumption that cave bear diet was based on C3 plants during the Pleistocene, and cave bear feeding behavior was unaffected by great climatic turnovers.

Acknowledgements We are thankful to our colleagues from Nicolaus Copernicus
, Poland) and Wroc1aw University (Wroc1aw, University (Torun Poland) for access to cave bear material from Bi
snik Cave. The authors are thankful to the reviewers for valuable advice and improvement of the earlier version of this manuscript. The research was supported by Polish National Centre for Science, grant number N N301 061540.

References Ábelová, M., Sabol, M., 2009. Attempt on the reconstruction of palaeotemperature and palaeoenvironment in the territory of Slovakia during the Last Glacial based on oxygen and carbon isotopes from tooth enamel and bone collagen of cave bears. Slovenský Kras e Acta Carsologica Slovaca 47 (Suppl. 1), 5e18. Altuna, J., 1973. Hallazgos de Oso Pardo (Ursus arctos, Mammalia) en cuevas del País Vasco. Munibe 25, 121e157. Ambrose, S.H., DeNiro, M.J., 1986. The isotopic ecology of East African mammals. Oecologia 69 (3), 395e406. Andrews, P., Turner, A., 1992. Life and death of the Westbury bears. Annales Zoologici Fennici 28, 139e149. Bocherens, H., 2004. Cave bear palaeoecology and stable isotopes: checking the rules of the game. In: Philippe, M., Argant, A., Argant, J. (Eds.), Proceedings of the 9th International Cave Bear Conference, Cahiers scientifiques du Centre de Conservation et d’Etude des Collections, Hors Série No 2. Muséum d’Histoire naturelle de Lyon, pp. 183e188. Bocherens, H., Fizet, M., Mariotti, A., 1994. Diet, physiology and ecology of fossil mammals as inferred from stable carbon and nitrogen isotope biogeochemistry: implications for Pleistocene bears. Palaeogeography, Palaeoclimatology, Palaeoecology 107 (3e4), 213e225. Bocherens, H., Fogel, M.L., Tuross, N., Zeder, M., 1995. Trophic structure and climatic information from isotopic signatures in a Pleistocene cave fauna of Southern England. Journal of Archaeological Science 22, 327e340. Bocherens, H., Koch, P.L., Mariotti, A., Geraads, D., Jaeger, J.-J., 1996. Isotopic biogeochemistry (13C, 18O) of mammalian enamel from African Pleistocene hominid sites. Palaios 11, 306e318. Bocherens, H., Stiller, M., Hobson, K.A., Pacher, M., Rabeder, G., Burns, J.A., Tütken, T., Hofreiter, M., 2011. Niche partitioning between two sympatric genetically distinct cave bears (Ursus spelaeus and Ursus ingressus) and brown bear (Ursus arctos) from Austria: Isotopic evidence from fossil bones. Quaternary International 245, 238e248. Bocherens, H., Fizet, M., Mariotti, A., Billiou, D., Bellon, G., Borel, J.-P., Simone, S., 1991. Biogéochimie isotopique (13C, 15N, 18O) et paléoécologie des ours pléistocènes de la Grotte d’Aldène (Cesseras, Hérault). Bulletin du Musée d’Anthropologie Préhistorique de Monaco 34, 29e49 (in French). Bryant, J.D., Froelich, P.N., 1995. A model of oxygen isotope fractionation in body water of large mammals. Geochimica et Cosmochimica Acta 59 (21), 4523e 4537. Cerling, T.E., Harris, J.M., 1999. Carbon isotope fractionation between diet and bioapatite in ungulate mammals and implications for ecological and paleoecological studies. Oecologia 120, 347e363. Chritz, K.L., Dyke, G.J., Zazzo, A., Lister, A.M., Monaghan, N.T., Sigwart, J.D., 2009. Palaeobiology of an extinct Ice Age mammal: stable isotope and cementum analysis of giant deer teeth. Palaeogeography, Palaeoclimatology, Palaeoecology 282, 133e144.

11

Cyrek, K., Socha, P., Stefaniak, K., Madeyska, T., Miros1aw-Grabowska, J., _ Sudo1, M., Czyzewski, q., 2010. The Palaeolithic of the Bi
snik Cave (Southern Poland) against the environmental background. Quaternary International 220, 5e30. DeNiro, M.J., 1985. Postmortem preservation and alteration of in vivo bone collagen isotope ratios in relation to palaeodietary reconstruction. Nature 317, 806e809. DeNiro, M.J., Epstein, S., 1978. Influence of diet on the distribution of carbon isotopes in animals. Geochimica et Cosmochimica Acta 42, 495e506. Dotsika, E., Zisi, N., Tsoukala, E., Poutoukis, D., Lykoudis, S., Giannakopoulos, A., 2011. Palaeoclimatic information from isotopic signatures of Late Pleistocene Ursus ingressus bone and teeth apatite (Loutra Arideas Cave, Macedonia, Greece). Quaternary International 245, 291e301. Drucker, D.G., Bocherens, H., Billiou, D., 2003. Evidence for shifting environmental conditions in Southwestern France from 33 000 to 15 000 years ago derived from carbon-13 and nitrogen-15 natural abundances in collagen of large herbivores. Earth and Planetary Science Letters 216, 163e173. Drucker, D.G., Bridault, A., Cupillard, C., Hujic, A., Bocherens, H., 2011. Evolution of habitat and environment of red deer (Cervus elaphus) during the Late-glacial and early Holocene in eastern France (French Jura and the western Alps) using multi-isotope analysis (d13C, d15N, d18O, d34S) of archaeological remains. Quaternary International 245, 268e278. Drucker, D.G., Bridault, A., Iacumin, P., Bocherens, H., 2009. Bone stable isotopic signatures (15N, 18O) as tracer of temperature variation in Lateglacial and early Holocene: case study of red deer from Rochedane site in French Jura. Geological Journal 44, 593e604. Fernández-Mosquera, D., Vila-Taboada, M., Grandal-d’Anglade, A., 2001. Stable isotopes data (d13C, d15N) from the cave bear (Ursus spelaeus): a new approach to its palaeoenvironment and dormancy. Proceedings of the Royal Society B: Biological Sciences 268 (1472), 1159e1164. Fricke, H.C., O’Neil, J.R., 1996. Inter- and intra-tooth variation in the oxygen isotope composition of mammalian tooth enamel phosphate: implications for palaeoclimatological and palaeobiological research. Palaeogeography, Palaeoclimatology, Palaeoecology (1e2), 91e99. García García, N., Feranec, R.S., Arsuaga, J.L., Bermúdez de Castro, J.M., Carbonell, E., 2009. Isotopic analysis of the ecology of herbivores and carnivores from the Middle Pleistocene deposits of the Sierra De Atapuerca, northern Spain. Journal of Archaeological Science 36, 1142e1151. Hercman, H., Gorka, P., 2002. Uranium-thorium analyses of fossil bones from Bi
snik Cave. In: Cyrek, K. (Ed.), Jaskinia Bi
snik. Rekonstrukcja zasiedlenia jaskini na tle zmian
srodowiska przyrodniczego. Wydawnictwo Uniwersytetu Miko1aja
, pp. 181e192 (in Polish with English summary). Kopernika, Torun Hilderbrand, G.V., Farley, S.D., Robbins, C.T., Hanley, T.A., Titus, K., Servheen, C., 1996. Use of stable isotopes to determine diets of living and extinct bears. Canadian Journal of Zoology 74, 2080e2088. Hillson, S., 2005. Teeth. Cambridge University Press, Cambridge, pp. 1e373. Kahlke, R.D., 1999. The History of the Origin, Evolution and Dispersal of the Late Pleistocene Mammuthus-Coelodonta Faunal Complex in Eurasia (Large Mammals). Fenske Companies, Rapid City. Koch, P.L., Fisher, D.C., Dettman, D., 1989. Oxygen isotope variation in the tusks of extinct proboscideans: a measure of season of death and seasonality. Geology 17, 515e519. Koch, P.L., Tuross, N., Fogel, M.L., 1997. The effects of sample treatment and diagenesis on the isotopic integrity of carbonate in biogenic hydroxylapatite. Journal of Archaeological Science 24, 417e429. Kohn, M.J., Schoeninger, M.J., Valley, J.W., 1996. Herbivore tooth oxygen isotope compositions: effects of diet and physiology. Geochimica et Cosmochimica Acta 60, 3889e3896. Kohn, M.J., Schoeninger, M.J., Valley, J.W., 1998. Variability on oxygen isotope compositions of herbivore teeth: reflections of seasonality or developmental physiology. Chemical Geology 152, 97e112. Krajcarz, M.T., 2009. Reconstruction of Sedimentary environments and Diagenesis of Pleistocene Sediments and Bone Remains from Bi
snik Cave (Polish Jura) on the Basis of Geochemical Analyses (Doctoral thesis). Faculty of Geology, University of Warsaw, Warszawa (in Polish). Krajcarz, M.T., Cyrek, K., 2011. The age of the oldest Paleolithic assemblages from Bi
snik Cave (southern Poland) in the light of geological data. Przegla˛ d Archeologiczny 59, 55e74. Krajcarz, M.T., Gola, M.R., Cyrek, K., 2010. Preliminary suggestions on the Pleistocene palaeovegetation around the Bi
snik Cave (Cze˛ stochowa Upland, Poland) based on studies of molecular fossils from cave sediments. Studia Quaternaria 27, 55e61. Krajcarz, M.T., Madeyska, T., 2012. Geology and chronostratigraphy of sediments from _ Bi
snik Cave. In: Cyrek, K., Czyzewski, q.A., Krajcarz, M.T. (Eds.), International Conference European Middle Palaeolithic during MIS 8 e MIS 3: Cultures e Environment e Chronology. Wolbrom, Poland, September 25the28th, 2012, pp. 56e58. Kurtén, B., 1968. Pleistocene Mammals of Europe. Aldine Transaction. New Brunswick and London. Lamparski, Z., 1961. The percentage distribution of local material in end-moraines of Middle-Polish Glaciation. In: Passendorfer, E. (Ed.), Prace o plejstocenie Polski
Srodkowej. Wydawnictwa Geologiczne, Warszawa, pp. 133e142 (in Polish with English summary). Lee-Thorp, J.A., Sponheimer, M., 2003. Three case studies used to reassess the reliability of fossil bone and enamel isotope signals for paleodietary studies. Journal of Anthropological Archaeology 22 (3), 208e216.

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022

12

M.T. Krajcarz, M. Krajcarz / Quaternary International xxx (2013) 1e12

Lewandowski, J., 1987. Odra glaciation in the Silesian Upland. Biuletyn Geologiczny 31, 247e312 (in Polish with English summary). Lewandowski, J., 2011. “Glacial oasis” in the Cracow-Cze˛ stochowa Upland e results of studies during the last fifty years. Przegla˛ d Geologiczny 59 (11), 732e738 (in Polish with English summary). Lindner, L., Marks, L., 2012. Climatostratigraphic subdivision of the Pleistocene Middle Polish Complex in Poland. Przegla˛ d Geologiczny 60 (1), 36e45 (in Polish with English summary). _ Lindner, L., Dzierzek, J., Lamparski, Z., Marks, L., Nitychoruk, J., 1995. An outline of the Pleistocene stratigraphy of Poland; main horizons of glacial deposits and their range (Zarys stratygrafii czwartorze¸du Polski; g1ówne poziomy osadów glacjalnych oraz ich rozprzestrzenienie). Przegla˛ d Geologiczny 43 (7), 586e591 (in Polish). Marciszak, A., 2012. Bloody canines and claws around the Bisnik Cave during OIS 8 _ e 3. In: Cyrek, K., Czyzewski, q.A., Krajcarz, M.T. (Eds.), The State of Research of Carnivore Assemblage Coexisted with Humans during the Last 200 Ka, International Conference European Middle Palaeolithic during MIS 8 e MIS 3: Cultures e Environment e Chronology. Wolbrom, Poland September 25the28th, 2012, p. 58. Marciszak, A., Socha, P., Nadachowski, A., Stefaniak, K., 2011. Carnivores from Bi
snik Cave. Quaternaire e Horssérie 4, 101e106. Marciszak, A., Stefaniak, K., 2010. Two forms of cave lion: Middle Pleistocene Panthera spelaea fossilis Reichenau, 1906 and Upper Pleistocene Panthera spelaea spelaea Goldfuss, 1810 from the Bi
snik Cave, Poland. Neues Jahrbuch für Geologie und Paläontologie e Abhandlungen 258 (3), 339e351. Mattson, D.J., 1998. Diet and morphology of extant and recently extinct northern bears. Ursus 10, 479e496. Michener, R., Lajtha, K. (Eds.), 2007. Stable Isotopes in Ecology and Environmental Science. Blackwell Publishing Ltd, Malden, Oxford e Carlton, pp. 1e566. Miros1aw-Grabowska, J., 2002. Geological value of Bi
snik Cave sediments (CracowCze˛ stochowa Upland). Acta Geologica Polonica 52 (1), 97e110. Mojski, J.E., 2005. Polish Lands in Quaternary. an Outline of Morphogenesis (Ziemie
stwowy Instytut GeoPolskie w Czwartorze˛ dzie. Zarys morfogenezy). Pan logiczny, Warszawa (in Polish). Nelson, B.K., DeNiro, M.J., Schoeninger, M.J., De Paolo, D.J., Hare, P.E., 1986. Effects of diagenesis on strontium, carbon, nitrogen and oxygen concentration and isotopic composition of bone. Geochimica et Cosmochimica Acta 50 (9), 1941e 1949. Nelson, D.E., Angerbjörn, A., Lidén, K., Turk, I., 1998. Stable isotopes and the metabolism of the European cave bear. Oecologia 116, 177e181. Pacher, M., Stuart, A.J., 2009. Extinction chronology and palaeobiology of the cave bear (Ursus spelaeus). Boreas 38 (2), 189e206. Passey, B.H., Robinson, T.F., Ayliffe, L.K., Cerling, T.E., Sponheimer, M., Dearing, M.D., Roeder, B.L., Ehleringer, J.R., 2005. Carbon isotope fractionation between diet, breath CO2, and bioapatite in different mammals. Journal of Archaeological Science 32, 1459e1470. Pérez-Rama, M., Fernández-Mosquera, D., Grandal-d’Anglade, A., 2011. Recognizing growth patterns and maternal strategies in extinct species using stable isotopes: the case of the cave bear Ursus spelaeus ROSENMÜLLER. Quaternary International 245, 302e306. Pushkina, D., Bocherens, H., Chaimanee, J., Jaeger, J.-J., 2010. Stable carbon isotope reconstructions of diet and paleoenvironment from the late Middle

Pleistocene Snake Cave in Northeastern Thailand. Naturwissenschaften 97, 299e309. Quade, J., Cerling, T.E., Barry, J.C., Morgan, M.E., Pilbeam, D.R., Chivas, A.R., LeeThorp, J.A., van der Merwe, N.J., 1992. A 16-Ma record of paleodiet using carbon and oxygen isotopes in fossil teeth from Pakistan. Chemical Geology 94 (3), 183e192. Rausch, R.L., 1969. Morphogenesis and age-related structure of permanent canine teeth in the brown bear, Ursus arctos L., in Arctic Alaska. Zeitschrift für Morphologie der Tiere 66 (2), 167e188. Reinhard, E., Torres, de T., O’Neil, J., 1996. 18O/16O ratios of cave bear tooth enamel: a record of climate variability during the Pleistocene. Palaeogeography, Palaeoclimatology, Palaeoecology 126, 45e59. Richards, M.P., Pacher, M., Stiller, M., Quilès, J., Hofreiter, M., Constantin, S., Zilhao, J., Trinkaus, E., 2008. Isotopic evidence for omnivory among European cave bears: late Pleistocene Ursus spelaeus from the Pestera cu Oase, Romania. Proceedings of the National Academy of Sciences USA 105, 600e604. _ Rózycki, S.Z., 1982. Roches moutonnèes and “exaration” basal moraines composed of local material in Ma1opolska Upland (Poland). Biuletyn Geologiczny 26, 107e 121 (in Polish with English summary). Schoeninger, M.J., DeNiro, M.J., 1982. Carbon isotope ratios of apatite from fossil bone cannot be used to reconstruct diets of animals. Nature 297, 577e578. Skrzypek, G., Wi
sniewski, A., Grierson, P.F., 2011. How cold was it for Neanderthals moving to Central Europe during warm phases of the last glaciation? Quaternary Science Reviews 30, 481e487. Sponheimer, M., Lee-Thorp, J.A., 1999. Alteration of enamel carbonate environments during fossilization. Journal of Archaeological Science 26, 143e150. Stiner, M.C., Achyuthan, H., Arsebük, G., Howell, F.C., Josephson, S., Juell, K., Pigati, J., Quade, J., 1998. Reconstructing cave bear paleoecology from skeletons: a cross disciplinary study of Middle Pleistocene bears from Yarimburgaz cave, Turkey. Paleobiology 24, 74e98. Torres, de T., 1988. Osos (Mammalia, Carnivora, Ursidae) del Pleistoceno Ibérico I. Filogenia; Distribución estratigráfica y geográfica. Estudio anatómico y métrico del cráneo. Boletín Geológico y Minero 99 (1), 3e46. Torres, de T.J., Quintero, I.A., Gomez, E.N., Mansilla, H.I., Martinez, C.D., 1978. Estudio comparativo de las mandíbulas de Ursus spelaeus, Rosemmuller e Heinrooth e Ursus deningeri, Von Reichenau y Ursus arctos, Linneo. Boletín Geológico y Minero 89 (3), 203e222. Vila Taboada, M., Fernández Mosquera, D., Grandal d’Anglade, A., 2001. Cave bear’s diet: a new hypothesis based on stable isotopes. Cuadernos do Laboratorio Xeoloxico de Laxe Coruña 26, 431e439. Wang, Y., Kromhout, E., Zhang, C., Xu, Y., Parker, W., Deng, T., Qiu, Z., 2008. Stable isotopic variations in modern herbivore tooth enamel, plants and water on the Tibetan Plateau: Implications for paleoclimate and paleoelevation reconstructions. Palaeogeography, Palaeoclimatology, Palaeoecology 260, 359e 374. Wiszniowska, T., Socha, P., Stefaniak, K., 2002. Quaternary fauna from sediments of Bi
snik Cave. In: Cyrek, K. (Ed.), Jaskinia Bi
snik. Rekonstrukcja zasiedlenia jaskini na tle zmian
srodowiska przyrodniczego. Wydawnictwo Uniwersytetu Miko1aja
, pp. 193e220 (in Polish with English summary). Kopernika, Torun Wright, L.E., Schwarcz, H.P., 1999. Correspondence between stable carbon, oxygen and nitrogen isotopes in human tooth enamel and dentine: infant diets at Kaminaljuyu. Journal of Archaeological Science 26, 1159e1170.

Please cite this article in press as: Krajcarz, M.T., Krajcarz, M., The 200,000 year long record of stable isotopes (d18O, d13C) of cave bear (Ursus spelaeus) teeth from Bi
snik Cave, Poland, Quaternary International (2013), http://dx.doi.org/10.1016/j.quaint.2013.07.022