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Tetracycline Labeled Bone Content Analysis of Ancient Nubian Remains from Kulubnarti

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Tetracycline Labeled Bone Content Analysis of Ancient Nubian Remains from Kulubnarti TETRACYCLINE LABELED BONE CONTENT ANALYSIS OF ANCIENT NUBIAN REMAINS FROM KULUBNARTI THESIS Presented in Partial Fulfillment of the Requirements for the Degree Master of Arts in the Graduate School of The Ohio State University By Julie Anna Margolis Graduate Program in Anthropology The Ohio State University 2015 Master's Examination Committee: Dr. Clark Spencer Larsen, co-advisor Dr. Samuel D. Stout, co-advisor Dr. Douglas E. Crews Copyrighted by Julie Anna Margolis 2015 ABSTRACT Armelagos and colleagues (2001) have hypothesized that beer is a conduit for in vivo tetracycline consumption by ancient Nubians. Streptomycetes bacteria has a high prevalence in Sudanese-Nubian soil (60 -70%) and secretes the antibiotic under harsh conditions such as fermentation. At the site of Kulubnarti, 21-S-46 cemetery (716 CE) skeletons likely represent a working underclass contemporaneous with the 21-R-2 cemetery (752 CE) containing the remains of a land-owning class. Interpretations of archaeological and osteological evidence suggest that poorer health and higher mortality occurred in the S population. To test whether an anticipated difference in tetracycline ingestion between S and R cemetery populations existed, the amount of tetracycline- labeled bone was quantified under ultra violet light using image analysis software. Amount of tetracycline labeling was expressed in terms of the total area of labeled bone tissue in square micrometers, number of labeled osteons, and number of grid intersections over labeled bone. No significant differences in percent tetracycline-labeled bone tissue, or percent labeled osteons was observed between cemeteries. These results suggest that tetracycline ingestion was similar for S and R group members, class differences were not mediating tetracycline ingestion, and both sub-groups had equal access to beer. ii Dedicated to my mentor, George J. Armelagos. iii ACKNOWLEDGMENTS This thesis would not have been possible without support from my parents, friends, and mentors. I would like to thank my committee co-chairs Dr. Samuel D. Stout and Dr. Clark S. Larsen, committee member Dr. Douglas E. Crews, and Dr. Mark Hubbe for their guidance and helpful comments regarding my writing and presentation of statistics. I am grateful to my friend Sara Becker (MSc. Maastricht University) for translating Pipenbrink’s 1983 work from German to English for me. I extend thanks to the Department of Anthropology at Emory University for allowing me to use their facilities and resources for data collection. I thank Nicole L. Henderson for having been an incredible lab assistant and for helping me test my proposed technique of image analysis by re-measuring several samples as part of a preliminary inter-observer study. Most of all, I would like to thank Dr. George J. Armelagos and Dr. Dennis P. Van Gerven, as without them this project would not have been possible. I am extremely grateful to Dennis for providing the Kulubnarti rib cross-sections for me to work with in the first place, helping me work through and understand my statistics, and answering a vast array of questions on his previous studies of ancient Nubia. As for George, I cannot express how thankful and appreciative I am for everything he did for me. George gave me this project to work on originally, gave me access to all of his equipment and resources (including funding for materials), answered my endless questions, acted as a iv sounding board for me to develop further research questions, and provided guidance as only a friend and mentor can in their ideal form. George prepared me for graduate school and a career in anthropology, and without him I would not be where I am today. I am also thankful that, while George did not see the final written version of this thesis before his death (only earlier drafts), he was able to see this project of ours through to the end of my analysis and the future directions of my research. I will be forever grateful to George Armelagos, for everything. v VITA 2008................................................................Diploma, Libertyville High School 2010................................................................A.A. Anthropology, Oxford College of Emory University 2012................................................................B.A. Anthropology, Emory University 2012 to present ..............................................Graduate Student, Department of Anthropology, The Ohio State University Publications Margolis JA, Van Gerven DP, and Armelagos GJ. 2013. Tetracycline labeling in early Christian burials from Kulubnarti, Nubia: Measure of class differences. American Journal of Physical Anthropology Supplement 56, 2013 (Annual Meeting Issue):189. Fields of Study Major Field: Anthropology vi Table of Contents ABSTRACT ........................................................................................................................ ii ACKNOWLEDGMENTS ................................................................................................. iv VITA .................................................................................................................................. vi Publications ........................................................................................................................ vi Fields of Study ................................................................................................................... vi Table of Contents .............................................................................................................. vii List of Tables ..................................................................................................................... ix List of Figures ..................................................................................................................... x CHAPTER 1: INTRODUCTION ...................................................................................... 1 Purpose ............................................................................................................................ 1 Background of Kulubnarti ............................................................................................... 2 Health of the Kulubnarti Populace .................................................................................. 5 Nubian Antibiotics .......................................................................................................... 7 Why Study Tetracycline Consumption at Kulubnarti? ................................................. 10 CHAPTER 2: MATERIALS AND METHODS .............................................................. 12 vii Materials ........................................................................................................................ 12 Methods ......................................................................................................................... 14 New Technique of Image Analysis Method .............................................................. 14 Osteon Count Method ................................................................................................ 17 Point-Count Method .................................................................................................. 17 CHAPTER 3: RESULTS .................................................................................................. 19 CHAPTER 4: DISCUSSION ............................................................................................ 24 CHAPTER 5: CONCLUSIONS ....................................................................................... 31 REFERENCES ................................................................................................................. 33 APPENDIX A: RAW DATA ........................................................................................... 37 APPENDIX B: RIB CROSS-SECTION PHOTOS .......................................................... 39 viii List of Tables Table 1: Sample specimens and associated sex and age estimations from Kulubnarti .... 13 Table 2: Percent averages of tetracycline-labeled bone for all ages at Kulubnarti ........... 20 Table 3: Percent averages of tetracycline-labeled bone among those aged 12 and older at Kulubnarti ......................................................................................................................... 23 Table 4: Kulubnarti cemetery 21-S-46 (S Cemetery) raw data ........................................ 37 Table 5: Kulubnarti cemetery 21-R-2 (R Cemetery) raw data…………………..………38 ix List of Figures Figure 1: Map of Nubia (Turner et al. 2007) ...................................................................... 3 Figure 2: Tetracycline molecule ......................................................................................... 8 Figure 3: Rib cross-section from specimen S48 before digital cleaning .......................... 15 Figure 4: Rib cross-section from specimen S48 after digital cleaning ............................. 16 Figure 5: Masks used for measurement of total bone area in microns (left) and tetracycline-labeled bone area in microns (right) for specimen S48 ................................ 17 Figure 6: Frequency of tetracycline-labeled bone area (in microns) by age for both cemeteries at Kulubnarti (loess line of fit included) ......................................................... 21 x CHAPTER 1: INTRODUCTION Purpose Ancient Nubians were ingesting the antibiotic, tetracycline, likely through beer (Bassett et al. 1980; Hummert and Van Gerven
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