Declaration Dr. Thomas H. Pringle - Buffalo Field Campaign

demonstrated that multiplex PCR can be efficiently employed for amplifying several loci
at once from fecal DNA samples, screening for sample quality, and genotyping individual
bison, while achieving low genotyping error rates. Microsatellite genotyping error rates
can be further reduced by screening many loci and using only those with low error rates,
(error rates varied among loci from 1% to 8%).
The per nucleotide error rate for mtDNA amplified from feces has not been
previously published for any species. For our mtDNA study, we detected an error rate of
0.0005 nucleotide mis-incorporations across all samples. This is approximately five times
higher than published error rates for high quality DNA sources with the Platinum taq we
used in our PCR amplification (0.0001) (Tindall and Kunkel 1988). We successfully
identified haplotypes for 120 of 127 individuals through mtDNA sequencing. Sequencing
of the mtDNA control region, and RFLP analysis, revealed two haplotypes among 179
bison sampled from both parks over two consecutive rut seasons. The 470 bp sequence
we amplified matched with the first 408 bp of haplotypes 6 and 8 previously defined by
Ward et al. (1999).
The frequencies of haplotypes 6 and 8 among the GYA breeding groups revealed
surprising strong evidence for genetic population structure. Significant differentiation
was detected between YNP and GTNP bison populations (FST = 0.191, p < 0.001).
However, the most intriguing result was the substantial genetic differences we found
among breeding groups within YNP. The differentiation between the Lamar Valley and
Hayden Valley was nearly twice that found between the parks (FST = 0.367, p < 0.001).
And, the differences in haplotype frequencies between Mirror Plateau and Hayden Valley
breeding groups were higher than between the parks as well (FST = 0.231, p < 0.001).
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