OJGen  Vol.3 No.3 , September 2013
Genetic analysis for geographic isolation comparison of brown bears living in the periphery of the Western Carpathians Mountains with bears living in other areas
ABSTRACT

Populations of the European brown bear (Ursus arctos L.) differ substantially in size, degree of geographic isolation and level of genetic diversity. Present patterns result from phylogeographic processes and profound human intervention. We assessed the genetic variability of a subpopulation of brown bears near the periphery of their range in the Western Carpathian Mountains and compared their genetic properties with those of bears in the core of the same population and elsewhere. Samples were collected non-invasively in 2007-2008 and2010 in Strázovské Vrchy Protected Landscape Area (PLA) in Slovakia (included in the NATURA 2000 networking programme). Seven polymorphic microsatellite loci (UaMU26, UaMU64, G10B, G1D, G10L, UaMU50 and UaMU51) were amplified using a nested PCR in order to assess the following parameters: variability, allelic combinations, heterozygosity, number of alleles and inbreeding coefficient. Sufficient brown bear DNA for analysis was obtained from 57 out of 140 samples (41%), among which 45 different genotypes were identified. Loci had a mean of 2.71 ± 0.76 alleles. Average observed heterozygosity was 0.59. The inbreeding coefficient was negative for all but one of the analysed loci (2007-2008). In the year 2010 was negative three of seven loci. These results imply that gene flow with other parts of the population has been maintained in the reduced level and the isolation level of bears in the study area was not so low. Nevertheless, the genetic variability of bears in Strázovské Vrchy PLA was lower than that reported from other localities in the Carpathian Mountains. The results are discussed in the context of behavioural ecology and conservation genetics.


Cite this paper
Graban, J. , Kisková, J. , Pepich, P. and Rigg, R. (2013) Genetic analysis for geographic isolation comparison of brown bears living in the periphery of the Western Carpathians Mountains with bears living in other areas. Open Journal of Genetics, 3, 174-182. doi: 10.4236/ojgen.2013.33020.
References
[1]   Swenson, J.E., Dahle, B., Gerstl, N. and Zedrosser, A. (2000) Action plan for the conservation of the brown bear in Europe (Ursus arctos)—Convention on the Conservation of European Wildlife and Natural Habitats (Bern Convention). Nature and Environment No. 114, Council of Europe Publishing, Strasbourg.

[2]   Sommer, R.S. and Benecke, N. (2005) The recolonization of Europe by brown bears Ursus arctos Linnaeus, 1758 after the Last Glacial Maximum. Mammal Review, 35, 156-164. doi:10.1111/j.1365-2907.2005.00063.x

[3]   Breitenmoser, U. (1998) Large predators in the Alps: The fall and rise of man’s competitors. Biological Conservation, 83, 279-289. doi:10.1016/S0006-3207(97)00084-0

[4]   Zedrosser, A., Dahle, B., Swenson, J.E. and Gerstl, N. (2001) Status and management of the brown bear in Europe. Ursus, 12, 9-12.

[5]   Zachos, F.E., Ottoa, M., Unici, R., Lorenzini and R., Hartl, G.B. (2008) Evidence of a phylogeographic break in the Romanian brown bear (Ursus arctos) population from the Carpathians. Mammalian Biology, 73, 93-101. doi:10.1016/j.mambio.2007.02.007

[6]   Randi, E., Gentile, L., Boscagli, G., Huber, D. and Roth, H.U. (1994) Mitochondrial DNA sequence divergence among some west European brown bear (Ursus arctos L.) populations. Lessons for conservation. Heredity, 73, 480-489. doi:10.1038/hdy.1994.146

[7]   Taberlet, P., Camarra, J.J., Griffin, S., Uhrès, E., Hanotte, O., Waits, L.P., Dubois-Paganon, C., Burke, T. and Bouvet, J. (1997) Noninvasive genetic tracking of the endangered Pyrenean brown bear population. Molecular Ecology, 6, 869-876. doi:10.1111/j.1365-294X.1997.tb00141.x

[8]   Lorenzini, R., Posillico, M., Lovari, S. and Petrella, A. (2004) Non-invasive genotyping of the endangered Apennine brown bear: A case study not to let one’s hair down. Animal Conservation, 7, 199-209. doi:10.1017/S1367943004001301

[9]   Taberlet, P. and Bouvet, J. (1994) Mitochondrial DNA polymorphism, phylogeography, and conservation genetics of the brown bear Ursus arctos in Europe. Proceedings of the Royal Society B: Biological Sciences, 255, 195-200.

[10]   Kohn, M., Knauer, F., Stoffella, A., Schroder, W. and Paabo, S. (1995) Conservation genetics of the European brown bear—A study using excremental PCR of nuclear and mitochondrial sequences. Molecular Ecology, 4, 95-103. doi:10.1111/j.1365-294X.1995.tb00196.x

[11]   Korsten, M., Ho, S.Y.W., Davison, J., Pahn, B., Vulla, E., Roht, M., et al. (2009) Sudden expansion of a single brown bear maternal lineage across northern continental Eurasia after the last ice age: A general demographic model for mammals? Molecular Ecology, 18, 1963-1979. doi:10.1111/j.1365-294X.2009.04163.x

[12]   Valdiosera, C.E., Garcia, N., Anderung, C., Dalen, L., Cregut-Bonnoure, E., Kahlke, R.-D., Stiller, M., Brandstrom, M., Thomas, M.G., Arsuaga, J.L., Gotherstrom, A. and Barnes, I. (2007) Staying out in the cold: Glacial refugia and mitochondrial DNA phylogeography in ancient European brown bears. Molecular Ecology, 16, 5140-5148. doi:10.1111/j.1365-294X.2007.03590.x

[13]   Swenson, J.E., Taberlet, P. and Bellemain, E. (2011) Genetics and conservation of European brown bears Ursus arctos. Mammal Review, 2, 87-98. doi:10.1111/j.1365-2907.2010.00179.x

[14]   Saarma, U., Ho, S.Y.W., Pybus, O.G., Kaljuste, M., Tumanov, I.L., Kojola, I., Vorobiev, A., Markov, N.I., Saveljev, A.P., Valdmann, H., Lyapunova, E.A., Abramov, A.V., Mannil, P., Korsten, M., Vulla, E., Pazetnov, S.V., Pazetnov, V.S., Putchkovskiy, S.V. and Rokov, A.M. (2007) Mitogenetic structure of brown bears (Ursus arctos L.) in northeastern Europe and a new time frame for the formation of European brown bear lineages. Molecular Ecology, 16, 401-413. doi:10.1111/j.1365-294X.2006.03130.x

[15]   Hartl, G.B. and Hell, P. (1994) Maintenance of high levels of allelic variation in spite of a severe bottleneck in population size: The brown bear (Ursus arctos) in the Western Carpathians. Biodiversity and Conservation, 3, 546-554. doi:10.1007/BF00115160

[16]   Jamnicky J. (1993) The hunt of the brown bear and European wolf in Slovakia a hundred years ago. Folia Venatoria, 23, 221-229. (in Slovak with English Summary)

[17]   Hell, P. and Slamecka, J. (1999) The bear in the Slovak Carpathians and in the world. PaRPress, Bratislava, 150p. (in Slovak).

[18]   Sabados, K. and Simiak, M. (1981) Distribution and hunting management of the brown bear (Ursus arctos L.) in Slovakia. Folia Venatoria, 10-11, 15-35. (in Slovak)

[19]   Rigg, R. and Adamec, M. (2007) Status, ecology and management of the brown bear (Ursus arctos) in Slovakia. Slovak Wildlife Society, Liptovsky Hrádok, 128p.

[20]   Jakubiec, Z. (2001) The brown bear Ursus arctos L. in the Polish part of the Carpathians. Polska Akademia Nauk, Kraków, 108p. (in Polish with English Summary)

[21]   Straka, M., Paule, L., Ionescu, O., Stofík, J. and Adamec, M. (2012) Microsatellite diversity and structure of Carpathian brown bears (Ursus arctos): Consequences of human caused fragmentation. Conservation Genetics, 13, 153-164. doi:10.1007/s10592-011-0271-4

[22]   Davison, J., Ho, S.Y.W., Bray, S.C., Korsten, M., Tammeleht, E., Hindrikson, M., Østbye, K., Østbye, E., Lauritzen, S. E., Austin, J., Cooper, A. and Saarma, U. (2011) Late-Quaternary biogeographic scenarios for the brown bear (Ursus arctos), a wild mammal model species. Quaternary Science Reviews, 30, 418-430. doi:10.1016/j.quascirev.2010.11.023

[23]   Paule, L., Krajmerová, D., Urban, P. and Adamec, M. (2006) Contribution to genetic diversity of brown bear (Ursus arctos L.) from the Western Carpathians. Vyskum a Ochrana Cicavcov na Slovensku, 7, 115-121. (in Slovak with English Abstract)

[24]   Janík, M., Voskár, J. and Buday, M. (1986) Present distribution of the brown bear (Ursus arctos) in Czechoslovakia. Folia Venatoria, 16, 331-352. (in Slovak with English Abstract)

[25]   Kruckenhauser, L., Rauer, G., Daubl, B. and Haring, E. (2009) Genetic monitoring of a founder population of brown bears (Ursus arctos) in central Austria. Conservation Genetics, 10, 1223-1233. doi:10.1007/s10592-008-9654-6

[26]   Brya, J. And Konecny, A. (2003) Fast sex identification in wild mammals using PCR amplification of the SRY gene. Folia Zoologica, 52, 269-274.

[27]   Bellemain, E. and Taberlet, P. (2004) Improved noninvasive genotyping method: application to brown bear (Ursus arctos) faeces. Molecular Ecology Notes, 4, 519-522. doi:10.1111/j.1471-8286.2004.00711.x

[28]   Janiga, M., Fecková, M., Kornan, J., Kalas, M., Labudíková, I., Matiasko, K. and Londa, P. (2006) Preliminary results on genetic tracking of the Brown Bear (Ursus arctos) individuals in the Malá Fatra National Park (Slovakia). Oecologia Montana, 15, 24-26.

[29]   Raymond, M. and Rousset, F. (1995) An exact test for population diferentiation. Evolution, 49, 1283-1286. doi:10.2307/2410454

[30]   Weir, B.S. and Cockerham, C.C. (1984) Estimating Fstatistics for the analysis of population structure. Evolution, 38, 1358-1370. doi:10.2307/2408641

[31]   Robertson, A. and Hill, W.G. (1984) Deviations from Hardy-Weinberg proportions: Sampling variances and use in estimation of inbreeding coefficients. Genetics, 107, 703-718.

[32]   Guo, S.W. and Thompson, E.A. (1992) Performing the exact test of Hardy-Weinberg proportion for multiple alleles. Biometrics, 48, 361-372. doi:10.2307/2532296

[33]   Goldstein, D.B. and Schlotterer, C. (1999) Microsatellites: Evolution and applications. Oxford University Press, Oxford.

[34]   Kimura, M. and Crow, J. (1964) The number of alleles that can be maintained in a finite population. Genetics, 49, 725-738.

[35]   Valdes, A.M., Slatkin, M. and Freimer, N.B. (1993) Allele frequencies at microsatellite loci: The stepwise mutation model revisited. Genetics, 133, 737-749.

[36]   Di Rienzo, A., Peterson, A.C., Garza, J.C., Valdes, A.M., Slatkin, M. and Freimer, N.B. (1994) Mutational processes of simple-sequence repeat loci in human populations. Proceedings of the National Academy of Sciences of the United States of America, 91, 3166-3170. doi:10.1073/pnas.91.8.3166

[37]   Marshall, T.C., Slate, J., Kruuk, L.E.B. and Pemberton, J.M. (1998) Statistical confidence for likelihood-based paternity inference in natural populations. Molecular Ecology, 7, 639-655. doi:10.1046/j.1365-294x.1998.00374.x

[38]   Saitou, N. and Nei, M. (1987) The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4, 406-425.

[39]   Zedrosser, A., StØen, O.-G., SæbØ, S. and Swenson, J.E. (2007) Should I stay or should I go? Natal dispersal in the brown bear. Animal Behaviour, 74, 369-376. doi:10.1016/j.anbehav.2006.09.015

[40]   Fernández, N., Selva, N., Yuste, C., Okarma, H. and Jakubiec, Z. (2012) Brown bears at the edge: Modelling habitat constrains at the periphery of the Carpathian population. Biological Conservation, 153, 134-142. doi:10.1016/j.biocon.2012.04.013

[41]   Find'o, S., Skuban, M. and Koren, M. (2007) Brown bear corridors in Slovakia. Carpathian Wildlife Society, Zvolen.

[42]   Pérez, T., Naves, J., Vázquez, J.F., Seijas, J., Corao, A., Albornoz, J. and Domínguez, A. (2010) Evidence for improved connectivity between Cantabrian brown bear subpopulations. Ursus, 21, 104-108. doi:10.2192/09SC018.1

[43]   Waits, L.P., Taberlet, P., Swenson, J.E., Sandegren, F. and Franzén, R. (2000) Nuclear DNA microsatellite analysis of genetic diversity and gene flow in the Scandinavian brown bear (Ursus arctos). Molecular Ecology, 9, 421-431. doi:10.1046/j.1365-294x.2000.00892.x

[44]   Tallmon, D.A., Bellemain, E., Swenson, J. and Taberlet, P. (2004) Genetic monitoring of Scandinavian brown bear effective population size and immigration. Journal of Wildlife Management, 68, 960-965. doi:10.2193/0022-541X(2004)068[0960:GMOSBB]2.0.CO;2

[45]   Paunovic, M. and Cirovic, D. (2006) Viability increase and recovery of brown bear (Ursus arctos L. 1758) population in northeastern Serbia—A Feasibility Study. Faculty of Biology. University of Belgrade, Belgrade.

[46]   Skrbinsek, T., Jelencic, M., Waits, L.P., Potocnik, H., Kos, I. and Trontelj, P. (2012) Using a reference population yardstick to calibrate and compare genetic diversity reported in different studies: An example from the brown bear. Heredity, 109, 299-305.

[47]   Kocijan, I., Galov, A., Cetkovic, H., Kusak, J., Gomercic, T. and Huber, D. (2011) Genetic diversity of Dinaric brown bears (Ursus arctos) in Croatia with implications for bear conservation in Europe. Mammalian BiologyZeitschrift fur Saugetierkunde, 76, 615-621.

[48]   De Barba, M., Waits, L.P., Garton, E.O., Genovesi, P., Randi, E., Mustoni, A. and Groff, C. (2010) The power of genetic monitoring for studying demography, ecology and genetics of a reintroduced brown bear population. Molecular Ecology, 19, 3938-3951. doi:10.1111/j.1365-294X.2010.04791.x

[49]   Paetkau, D., Waits, L.P., Clarkson, P., Craighead, L., Vyse, E.R., Ward, R. and Strobeck, C. (1998) Variation in genetic diversity across the range of North American brown bears. Conservation Biology, 12, 418-429.

[50]   Tammeleht, E., Remm, J., Korsten, M., Davidson, J., Tumanov, I., Saveljev, A., Mannil, P., Kojola, I. and Saarma, U. (2010) Genetic structure in large, continuous mammal populations: The example of brown bears in northwestern Eurasia. Molecular Ecology, 19, 5359-5370. doi:10.1111/j.1365-294X.2010.04885.x

[51]   Aarnes, S.G., Bellemain, E., Eiken, H.G. and Wartainen, I. (2009) Interlaboratory comparison of genetic profiles of brown bears from Sweden (Laboratoire d’Ecologie Alpine) and Norway (Bioforsk Svanhovd). Bioforsk Report, 4, 133.

[52]   Karamanlidis, A.A., De Barba, M., Georgiadis, L., Groff, C., Jelincic, M., Kocijan, I., Kruckenhauser, L., Rauer, G., Sindicic, M., Skrbinsek, T. and Huber, D. (2009) Common guidelines for the genetic study of brown bears (Ursus arctos) in southeastern Europe. LCIE, Athens.

 
 
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