Enderby Island Cattle: What Breed Are They Derived From?
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Proc. Assoc. Advmt. Anim. Breed. Genet. Vol 14 ENDERBY ISLAND CATTLE: WHAT BREED ARE THEY DERIVED FROM? D.L. Hyndman1, R.P. Littlejohn1, J.L. Williams2 and A.M. Crawford3 1AgResearch, Invermay Agricultural Centre, Private Bag 50034, Mosgiel, New Zealand. 2Roslin Institute, Midlothian, Edinburgh, EH25 9PS, Scotland. 3AgResearch, Molecular Biology Unit, Biochemistry Dept. University of Otago, Box 56, Dunedin, New Zealand. SUMMARY In the summer of 1993 the last cattle were removed from sub-antarctic Enderby Island where they had remained isolated since at least 1896. We have examined the DNA from 9 of the last surviving animals to determine which of the modern breeds they are most closely related to. The genotypes of each individual were determined at 20 different loci and the data submitted to the Genetic Diversity Database at Roslin Institute, Scotland. (http://www.ri.bbsrc.ac.uk/cdiv_www/homepage.htm). Allele frequency distributions were compared with those of a variety of British breeds and the Danish Shorthorn. Of the breeds examined the Jersey was the most closely related to the Enderby Island cattle. Keywords: Microsatellite, Allele frequency, Bos taurus, Enderby Island, Auckland Islands. INTRODUCTION Enderby Island is a small island (800 ha) that is part of the Auckland Islands. These are isolated sub- Antarctic islands lying in the “furious fifties” approximately 400 km south of the New Zealand mainland. Early attempts at settlement in the 1840’s and early 1850’s failed but did include the release of cattle, sheep, goats, dogs, pigs, rabbits and, unintentionally, mice at this time. (Taylor 1971). Although settlement was abandoned by the English owned Southern Whale Fishery Company in 1852, a Maori settlement begun in 1842 remained for a further 4 years until they left the island in 1856. Although cattle were left on the island by the departing English in 1852 there is only one report of visitors seeing cattle on Enderby Island between 1852 and 1894. This was in 1873 when HMS Cossack visited the island. All other visitors including shipwrecked sailors who stayed on the island for extended periods did not find cattle. It is assumed that the early cattle died out soon after 1852 and that Dr F.A. Monkton, who held the pastoral lease in 1873 was responsible for the sighting of cattle in 1873. Despite many visits to the island during the 1880’s no cattle were reported on the island. The final introduction of cattle occurred during the years 1894 to 1896 when Mr Moffet of Invercargill, who held the pastoral lease, landed an unknown number of cattle in 1894, 3 animals in 1895 and 9 animals in 1896. He used the NZ government steamer Hinemoa, which regularly visited the islands during this period. Since then all visitors to the island observed the presence of cattle and there have been no further introductions. 329 Proc. Assoc. Advmt. Anim. Breed. Genet. Vol 14 In order to better preserve the unique biota of these islands the NZ Government began a programme for the eradication of introduced mammals. Cattle were eradicated on Enderby Island over the summers of 1991 to 1993. Thanks to the efforts of the Rare Breeds Conservation Society of NZ and the NZ Department of Conservation 800 straws of semen from Enderby island bulls were collected. A single female cow was also rescued but she proved infertile. This rare animal was then cloned and 5 clones are currently being grown to adulthood. They will provide the foundation dams for the continued survival of these rare animals. Microsatellite allele frequency distributions are now the accepted way of measuring the genetic distance between breeds of animals (Buchanan et al. 1994). In order that all breeds can be compared with one another an agreement between researchers to use a common set of markers is essential. To expedite this the Genetic Diversity Database (http://www.ri.bbsrc.ac.uk/cdiv_www/homepage.htm) was set up at Roslin Institute, Scotland with funding from the European Commission. They also provide a set of bovine DNA standards of known allele size so that all contributors can standardise their microsatellite data. This paper describes our genotyping of the Enderby Island cattle and the subsequent comparison with other genotypes from seven other breeds to determine which of these they most closely resemble. METHODS DNA purification The semen of 9 bulls collected by the Rare Breeds Conservation Society of NZ with the assistance of Department of Conservation during the culling of the Enderby Island cattle in 1993 was made available for the study. The DNA was extracted from frozen semen using the following method. Semen straws were thawed and the fluid transferred to a 1.5ml Eppendorf tube. The sperm were washed by two cycles of pelleting and resuspension in 1 X PBS. The final pellet was then resuspended in 0.5 ml of 0.01M Tris HCl (pH 7.2), 0.01M NaCl, 0.01M EDTA, 0.4% SDS. To the suspension was added 15ml of Mercaptoethanol, and 20mg Proteinase K, which was then incubated overnight at 55oC. The DNA was then precipitated by adding 50ml of a saturated NaCl solution and 1ml of 95% ethanol. The precipitated DNA was pelleted, washed once with 70% ethanol and redissolved in 200ml of TE buffer. Genotyping The Roslin Institute provided DNA from 6 cattle for reference to ensure that the microsatellite allele genotypes determined for the Enderby Island samples were consistent with those in the Cattle Diversity database. All nine Enderby Island bulls were genotyped with the following microsatellite markers. INRA063, INRA005, ETH225, ILSTS005, HEL5, HEL1, INRA023, ETH10, HEL9, CSSM66, INRA032, ETH3, BM2113, BM1824, HEL13, INRA037, BM1818, ILSTS006, MM12, CSRM60, ETH185, HAUT24, HAYT27, TGLA227, TGLA126, TGLA122, TGLA53, SPS115. Each locus was amplified using PCR in which one of the primers had been end labelled with 33P and the product size (alleles) determined using 6% poly acrylamide gel electrophoresis as previously described (Crawford et al. 1991). 330 Proc. Assoc. Advmt. Anim. Breed. Genet. Vol 14 Statistical analysis The coefficient of coancestry (q) (Weir 1996) was calculated pairwise between Enderby Island cattle and each other breed, for all microsatellites for which data were mutually available. The 95% confidence intervals were based on 1000 bootstrap samples across loci, and were calculated using GDA (Lewis and Zaykin 2001). RESULTS AND DISCUSSION Estimates of q, together with 95% confidence intervals, are presented in Table 1. The upper bound of the confidence interval for Jerseys of 0.2680 is less than the lower bound for any of the other breeds. While this is not conclusive evidence that Enderby Island cattle originated from Jersey stock, it practically eliminates the other 6 breeds as contenders. No genotype data were available on British Shorthorns, which are the breed that anecdotal evidence (Falla 1948) suggests they were derived from. We did however have access to the data from the Danish Shorthorn breed. The q value for this breed indicated that they were not likely to be the breed of origin. Table 1: Estimates of the Coefficient of Co-ancestry (q) between Enderby Island cattle and six other European breeds Breed Angus Belted Friesian Hereford Jersey Shetland Danish Galloway Shorthorn q 0.3847 0.4763 0.3592 0.3625 0.2024 0.5577 0.4564 95% lower 0.3403 0.3902 0.2881 0.2991 0.1440 0.4385 0.3069 bound 95% upper 0.4307 0.5732 0.4292 0.4367 0.2680 0.6726 0.5981 bound ACKNOWLEDGEMENTS The authors wish to thank the Rare Breeds Conservation Society of NZ in particular Mr D. Matheson, Mr M Willis and Prof. H Blair for the support and encouragement we have received. We also thank Dr D. Wells for providing the semen from the Enderby Island cattle and Geraldine Russell for help in using the Genetic Diversity Database. We are indebted to Drs, L.E. Holm, (Danish Shorthorn) M.L. Glowatski (Jersey) and D. Bradley (Jersey) in addition to J.L.Williams (all other breeds including Jersey) for submission of the genotype information we used from the database. REFERENCES Buchanan, F.C., Adams, L.J., Littlejohn, R.P., et al. (1994) Genomics 22: 397. Crawford A.M., Buchanan, F.C. and Swarbrick, P.A. (1991). Proc. N.Z. Soc. Anim. Prod. 51: 79. Falla, R.A. (1948) NZ Geographer. 4:127. Lewis, P.O. and Zaykin, D. (2001) Genetic Data Analysis: Version 1.0 (d16c). Free program available from “http://lewis.eeb.uconn.edu/lewishome/software.html” Taylor R.H. (1971) N.Z. J. Botany 9: 225. Weir, B.S. (1996) Genetic Data Analysis. Sinauer Associates, Inc; Sunderland, Massachusetts. 331 Proc. Assoc. Advmt. Anim. Breed. Genet. Vol 14 332 .