ABO Blood Group Frequencies in Pre-European Contact America: An Ancient DNA Analysis

1. ABO Blood Group Frequencies in Pre-European Contact America: An Ancient DNA Analysis Melissa Halverson M.A. Thesis, May 2006

2. Outline • European Contact (French, Spanish, English) • Smallpox • European-induced genetic bottleneck? • Peopling of the Americas (genetic diversity, founding populations); genetic bottleneck? • Hypothesis/Results/Conclusion • Future Work

3. European Contact- the French • Controlled trade • Integrated Native American culture and technology into their lifestyle • Took Native Americans to France • Native Americans became reliant on French supplies • Native Americans fell under colonial government Libraries and Archives Canada

4. European Contact- the Spanish • Enslaved Native Americans in Florida • Christian missionization • mestizaje mix of Spanish, Africans, Native American • Fundamentally a new multi-ethnic society • New plants and animals Blessed Kateri, clip art

5. European Contact- the English • English moved Native Americans off land • Farms of “higher use” • Boundary confrontations • Crown permission to exterminate • Maintain English culture • Basis for Removal Act (Trail of Tears)

6. Consequences of Contact • Warfare • Slavery • Relocations • Disease • Depopulation

7. Consequences of Contact • Warfare • Slavery • Relocations • Disease • Depopulation *before 1500 A.D. = 1-18 million (Ubelaker 1988; Dobyns 1983) *1500 A.D. to 1900 = 530,000 (Dobyns 1983)

8. Disease Example: Outline of Florida Epidemiology 1519 Smallpox 1545-1548 Bubonic Plague 1549 Typhus 1550 Mumps 1559 Influenza 1613-1617 Bubonic Plague 1649 Yellow Fever 1653 Smallpox 1659 Measles 1727 Timucuan Extinction (Dobyns 1983) “Pox Americana, “ Elizabeth Fenn

9. Smallpox Watts 2003

10. Non-infectious incubation period (12 days) • Fever up to 104°F, back pain, vomiting • Rash on face, arms, hands, trunk • Lesions ulcerate in mouth, releasing virus down throat, suffocating individual • Pustules form scabs 8-14 days later *Spread through respiratory contact until last scabs fall off (WHO 2006; Trimble 1979) medinfo.ufl.edu

11. Mortality Rates • Eurasia = ~30% • “Virgin soil” Native Americans = ~38.5% Aztecs ~50% Piegan, Huron, Catawba, Cherokee, and Iroquois ~66% Omaha, Blackfeet ~90% Mandan ~100% Taino Indigenous Perspectives • Supernatural forces, animal spirits • Punishment for violations of tribal laws • Witchcraft • Jesuits/missionaries • Europeans

12. Treatments, Responses, and Effects • Sweat lodge with herbs and cold water plunging • Avoiding diseased villages • Smallpox dance (the Ahtawhungnah) (Cherokee) • Quarantining • Inoculation/vaccination • Suicide • Abandonment • Mourning Wars (Iroquois) • Losses hurt subsistence, defense, and cultural roles • Fusion of neighboring groups • Decreased fertility (pop loss, impotence, poxmarks, blindness)

13. Wang et al. (2004) - Oklahoma Choctaw heterozygosity deficiency, reduced quantity of segregating alleles, high frequency of multiple sclerosis • Bolnick and Smith (2003) – Southeastern populations have low haplotype diversity, nucleotide diversity, and high level of haplotype extinction • Ribeiro-dos-Santos et al. (1996) – Pre-contact Brazilian Amazonians have high percentage of non-A-D haplogroups than extant groups *European-induced genetic bottleneck

14. Peopling of the Americas • 14 kya either through Beringia and western coast of Americas (Mandryk et al. 2000)

15. Haplogroups A-D, X • Found in North, Central, and South America • Encompass 96-100% of extant Native American mitochondrial haplotypes Eshleman et al. 2003

16. Small founding population(s) from south-central Siberia (Smith et al. 1999) Schurr 2004 • A-D and X brought in single migration from Siberia • A-D brought by initial migration, X is separate migration • A, C, D brought by initial migration, B and X are separate and with different geographic origins • C and D reintroduced in second migration.

17. Evidence for genetic bottleneck • 20x increase in restriction patterns in Amerinds compared to 2% found in Asians (Wallace and Torroni 1992; Schurr et al. 1990) • Founder size of 2000 females (Wallace et al. 1985); 70 reproductive individuals (Hey 1997)

18. North and South America- 90% O • Europe- 25-55% A • Australian aborigines- 45% A • Central Asia- 20-30% B ABO Blood Group Smallpox Associations • Molecular mimicry- Substance similar to A antigen in strain of smallpox (Vogel, Pettenkofer, and Helmbold 1960) • Higher disease severity, greater smallpox scarring in A and AB (Vogel 1965) • Asian epidemics- 115/200 infected had A and AB types (Chakravartti et al. 1966) • 60% selective disadvantage against AA, 36% AO and AB, 0% B and O (Adalsteinsson 1985)

19. Hypothesis • Is the high frequency of blood type O in extant Native Americans the product of a genetic bottleneck that occurred during the initial peopling of the Americas or one that occurred following European contact?

20. Methods • DNA extracts from Pete Klunk Mound (1825 +/- 75 YBP) and Wright Mound (2190-1790 YBP)-represent pre-contact • Extant frequencies pulled from literature • PCR using ABO primers • RFLP analysis • Phenotype (extant), genotype, allelic diversity compared

21. PCR * Successfully amplified ancient nuclear DNA!

22. RFLP Analysis (Restriction Fragment Length Polymorphism)

23. Hummel et al. 2002

24. Hummel et al. 2002

25. Hummel et al. 2002

26. Results • Both pre-contact/extant populations had 90% type O blood • No statistically significant differences (P=1.000) • OivOiv genotype most common (50%) • No Hardy-Weinberg equilibrium (P= 0.004) • Klunk Mounds – individuals most likely belonged to the same population

27. Conclusion • First study to use ancient DNA to ascertain ABO blood types of prehistoric Native Americans • Low type A frequency may stem from a genetic bottleneck that occurred during initial peopling of the Americas • The large population decline during the European contact period did not significantly impact the genetic distribution of ABO blood types

28. Plans Before Publication • Find genotype/allele diversity in extant Native American populations (will use extracted DNA) • Further prehistoric blood typing from Western US, Canada, and South America • Further contamination prevention (STR) (AmpFlSTR Profiler Plus forensic kit)

29. Acknowledgements • Drs. Deborah Bolnick, Sam Wilson, and John Kappelman • Chickasaw, Shawnee, and Cherokee nations • The Bolnick Lab • IPAS • Daniel Perrault • Dr. Della Cook • Dr. George Crothers and the W.S. Webb Museum of Anthropology • University of Texas College of Liberal Arts • Elsevier and Springer-Verlag Publishing