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caraccio2003malasia

caraccio2003malasia

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Published by Karumbé

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Published by: Karumbé on Jun 14, 2011
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Caraccio M. N., A. Formia, M. Hernandez, A. Fallabrino and M. Bruford. (In press). Preliminary Mixed Stock Analysis of  Juvenile Green Turtles in Uruguay using Mitochondrial Dna Sequences. Proceedings of the 23° International Symposiumon Sea Turtle Biology and Conservation, Kuala Lumpur, Malasia. U.S. Dep. Commer. NOAA Tech. Mem (2003).
PRELIMINARY MIXED STOCK ANALYSIS OF JUVENILE GREEN TURTLESIN URUGUAY USING MITOCHONDRIAL DNA SEQUENCES
Maria Noel Caraccio
1,3
, Angela Formia
2
, Martin Hernandez
1,3
,Alejandro Fallabrino
1
and Michael W Bruford
2
 
1C.I.D., Proyecto Karumbé, Tortugas Marinas del Uruguay , J. Paullier 1198, Montevideo, Uruguay,mnoel@fcien.edu.uy2School of Biosciences, Cardiff University, Cardiff CF10 3TL, United Kingdom3Sección Genética Evolutiva, Facultad de Ciencias. Iguá 4225, Montevideo, Uruguay
INTRODUCTION
The green turtle,
Chelonia mydas
, similarly to other sea turtle species, has a life cycle that is hard tostudy directly due to the difficulty of tracking migratory movement among feeding and nestingareas, and to the long maturation times. After reaching the sea, the hatchlings are not seen againuntil they enter neritic foraging grounds as juveniles. These areas often contain mixed stocks(individuals belonging to more than one nesting population) and are located many kilometers awayfrom the nesting grounds. Studies carried out in nesting areas indicate that adult females return tonest at their natal beach. Most of the knowledge concerning sea turtle distribution is obtained fromtagging experiments. While nesting females are easily accessible on the beach, feeding groundsrequire greater efforts, including catching individuals at sea with tangle nets. For some years,Karumbé Project has been monitoring the feeding grounds of juvenile green turtles along theUruguayan coast, and performing activities such as capturing and tagging turtles through netting,and locating strandings or incidental fishery catches. Recently, tissue sampling has been carried outin order to determine the genetic characteristics of green turtles inhabiting Uruguayan waters. Thecontrol region of mitochondrial DNA (mtDNA) is a molecular tag commonly used inphylogeographic studies of this species in the Atlantic, contributing to reveal their molecularevolution, population structure, reproductive behavior and migratory patterns (Bowen et al. 1992,Allard et al. 1994, Lahanas et al. 1994, Encalada et al. 1996, Lahanas et al. 1998). MitochondrialDNA evidence can also be used to define management units for conservation (Moritz 1994). In thiswork we present the preliminary results of analysis carried out in the Biodiversity andEnvironmental Process Research Group at Cardiff University (UK) using the control region of mtDNA of juvenile green turtles found along the Uruguayan coast.
MATERIALS AND METHODS
Tissue samples from twenty juvenile green turtles were collected from stranded individuals, fromturtles that were captured by netting efforts, and from turtles incidentally caught by artisanalfisheries. The size of the turtles ranged between 32 and 52cm of curved carapace length notch to tip(CCLn-t) (Bolten 1999). The samples were preserved in absolute ethanol. Extraction of the mtDNAwas perfomed using a modification of the protocol by Allen et al. (1998). A fragment of 486 bpfrom the control region was amplified with the primers HDCM1 and LDCM1 (Allard et al. 1994),and HDCM1.1 and LDCM1.1 for degraded DNA (Formia 2002). PCR products were visualised on1.5% agarose gels and were later sequenced using automated sequencing protocols with anABI3100 Sequencer. The sequences obtained were aligned using SEQUENCHER software (GeneCodes Corporation) and analysed using ARLEQUIN (Schneider et al. 2000). Also, mixed stock analyses were carried out with BAYES (Pella & Masuda 2001), to estimate the proportion of nesting populations that contributed to our juvenile stock.
 
Caraccio M. N., A. Formia, M. Hernandez, A. Fallabrino and M. Bruford. (In press). Preliminary Mixed Stock Analysis of  Juvenile Green Turtles in Uruguay using Mitochondrial Dna Sequences. Proceedings of the 23° International Symposiumon Sea Turtle Biology and Conservation, Kuala Lumpur, Malasia. U.S. Dep. Commer. NOAA Tech. Mem (2003).
RESULTS
Five polymorphic sites were found in the twenty samples, defining six haplotypes already describedfor nesting beaches of the Atlantic Ocean: CM5 (previously detected in Mexico, Aves Island,Surinam and Sao Tome), CM6 (also found in Bioko, Ascension Island,Sao Tome, Suriname), CM8(Bioko, Ascension Island, Guinea Bissau, Sao Tome, Principe and Atol das Rocas), CM9(Ascension Island and Atol das Rocas), CM10 (Ascension Island) and CM24 (Ascension Island)(Lahanas et al. 1994, Encalada et al. 1996, Lahanas et al. 1998, Formia 2002). Eleven of the twentyindividuals exhibited the CM8 haplotype, five individuals had CM5, and CM6, CM9, CM10 andCM24 haplotypes were each represented once. Haplotypic diversity was found to be 0.6579 +/-0.0957, comparable to values for other mixed stocks (Formia 2002) and indicating relatively highgenetic variation. The results of mixed stock analyses (using the nine baseline rookeries listedabove as potential contributors, Figure 1) showed that the nesting population which contributedmost to the feeding ground of Uruguay was the Ascension Island rookery with a mean of 62%followed by, Matapica (Surinam) with 14% and Aves Island (Venezuela) with 12% (see Table 1).The lower confidence limit for Ascension Island (above zero) confirmed that it was definitelypresent among the Uruguayan green turtle stock.
DISCUSSION
When analyzing these preliminary results, we have to be aware of the number of samples used andthe incomplete descriptions available for several of the potentially contributing rookeries. Resultsclearly showed the necessity for further studies based on greater sample sizes. Despite this fact, thesequences obtained from these juvenile greens provided enough information to presume that thecoast of Uruguay is an area hosting green turtles from several nesting beaches in the AtlanticOcean. The presence of 6 different haplotypes in only 20 samples, showed this recruitment zone for juvenile greens to be genetically diverse. The results obtained using BAYES suggested thatAscension Island is likely to be the greatest contributor to the Uruguayan genetic stock (a mean of 
 
Caraccio M. N., A. Formia, M. Hernandez, A. Fallabrino and M. Bruford. (In press). Preliminary Mixed Stock Analysis of  Juvenile Green Turtles in Uruguay using Mitochondrial Dna Sequences. Proceedings of the 23° International Symposiumon Sea Turtle Biology and Conservation, Kuala Lumpur, Malasia. U.S. Dep. Commer. NOAA Tech. Mem (2003).
62%). However, we cannot exclude other areas, particularly the large rookery of Trindade, forwhich genetic data is not yet available.A comprehensive study and increased sample sizes, could elucidate some unresolved questionsabout origin, distribution, dispersal movements and migratory behavior of these green turtlepopulations. We hope that a comprehensive genetic survey will aid in developing appropriatemanagement strategies for the endangered green turtles in the region.
ACKNOWLEDGEMENTS
We are grateful to the British Embassy in Uruguay for financial support of the research at Cardiff University. Thank to British Petroleum Conservation Programme, National Fish and WildlifeFoundation for supporting Karumbe Project.We would also like to thank Karumbe's friends for collecting the samples and the David and LucillePackard Foundation for supporting our participation at the Symposium.Scientific and collection permit : Nº 270/01 (Dirección General de Recursos Naturales Renovables).CITES permit Uruguay : 000137
REFERENCES
Allard M, Miyamoto MM, Bjorndal KA, Bolten AB, Bowen BW (1994) Support for natal homingin green turtles from mitochondrial DNA sequences. Copeia 1994:34-41Allen M, Engstrom AS, Meyers S et al. (1998) Mitochondrial DNA sequencing of shed haris andsaliva on robbery caps: sensitivity and matching probabilities. Journal of Forensic Sciences 43:453-464.Bowen BW, Meylan AB, Ross JP, Limpus CJ, Balaz GH, Avise JC (1992) Global populationstructure and natural history of green turtle (Chelonia mydas) in terms of matriarchal phylogeny.Evolution 46:865-881Bolten AB (1999) Techniques for Measuring Sea Turtles pp. 110-114. In: Eckert KL., Bjorndal KA,Abreu-Grobois FA and Donnelly M (Editors). Research and Management Techniques for theConservation of Sea Turtles. IUCN/SSC Marine Turtle Specialist Group Publication N° 4Encalada S, Lahanas PN, Miyamoto MM, Bjorndal KA, Bolten AB, Bowen BW (1996)Phylogeography and population structure of the Atlantic and Mediterranean green turtle Cheloniamydas: a mitochondrial DNA control region sequence assesment. Molec. Ecol. 5: 473-483Formia A (2002) Population and genetic structure of the green turtle (Chelonia mydas) in West andCentral Africa: implications for management and conservation. PhD Thesis, Cardiff University,UK.Lahanas PN, Miyamoto MM, Bjorndal KA, Bolten AB (1994) Molecular evolution and populationgenetics of Greater Caribbean green turtles (Chelonia mydas) as inferred from mitochondrial DNAcontrol region sequences. Genetica 94:57-67Lahanas PN, Bjorndal KA, Bolten AB, Encalada SE, Miyamoto MM, Valverde RA and Bowen BW(1998) Genetic composition of a green turtle (Chelonia mydas) feeding ground population: evidencefor multiple origins. Marine Biology 130: 345-352

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