Development of microsatellite DNA sequencies for the study of remnant populations of Broad-snouted caiman (Caiman latirostris), of central region of Sao Paulo State

Novos marcadores genéticos foram caracterizados para jacaré-de-papo-amarelo (Caiman latirostris) pela construção de bibliotecas enriquecidas de DNA microssatélite. Um microssatélite foi desenvolvido a partir de uma biblioteca enriquecida de DNA microssatélite (ACC/TGG)n e 12 a partir de uma biblioteca enriquecida de DNA microssatélite (AC/TG)n. Esses marcadores foram testados em indivíduos da espécie Caiman latirostris e resultaram em sete novos microssatélites polimórficos. Adicionalmente quatro marcadores microssatélites de Alligator mississipiensis previamente transferidos para Caiman latirostris foram utilizados. Amostras de sangue jacarés-de-papo-amarelo originárias de várzeas associadas ao Rio Piracicaba e alguns de seus tributários no estado de São Paulo, Brasil, foram avaliadas quanto à variação genética entre populações e o parentesco entre indivíduos. Foi detectada variabilidade entre indivíduos originários de sitos diferentes, mesmo entre aqueles com pequena distância geográfica. Os resultados sugerem que os grupos amostrados em cada sítio são compostos predominantemente por indivíduos aparentados. Uma possível combinação de alta taxa de mortalidade e baixa taxa de natalidade pode ser a explicação do baixo número de indivíduos dispersos com sucesso por geração entre os sítios estudados. Esses marcadores podem auxiliar na compreensão dos processos metapopulacionais que aparentemente ocorrem na espécie.New genetic markers were characterized for the broad-snouted caiman (Caiman latirostris) by constructing libraries enriched for microsatellite DNA. One microsatellite was developed from a (ACC/TGG)n enriched microsatellite DNA library and 12 from a (AC/TG)n enriched microsatellite DNA library. These markers were tested in Caiman latirostris individuals and resulted in seven new polymorphic microsatellites for the specie. Additionally four Alligator mississipiensis microsatellite markers previously transferred for Caiman latirostris were used. Samples from broad-snouted caimans from small wetlands associated with the Piracicaba River and some of its tributaries in the state of São Paulo, Brazil were used to study the genetic variation between populations and parentage between individuals. Genetic variability was detected among individuals from different sites, even those within a small geographic distance. The results suggest that the groups sampled at each site are composed predominantly of related individuals. A possible combination of high mortality and low natality rates in the fragmented Caiman latirostris populations may explain the low number of successfully dispersed individuals per generation observed between the sites studied. These markers might help to understand the metapopulation processes that are occurring within this species

Cross-species microsatellite amplification in South American Caimans (Caiman spp and Paleosuchus palpebrosus)

Microsatellite DNA markers have been used to assess genetic diversity and to study ecological behavioral characteristics in animals. Although these markers are powerful tools, their development is labor intensive and costly. Thus, before new markers are developed it is important to prospect the use of markers from related species. In the present study we investigated the possibility of using microsatellite markers developed for Alligator mississipiensis and Caiman latirostris in South American crocodilians. Our results demonstrate the use of microsatellite markers for Paleosuchus palpebrosus, Caiman crocodilus and Caiman yacare

Effective population size of broad-snouted caiman (Caiman latirostris) in Brazil: A historical and spatial perspective

Caiman latirostris has a large geographic distribution, that includes Argentina, Bolivia, Brazil, Paraguay, and Uruguay. In Brazil illegal hunting and land use change have caused population decline, relatively well documented in the last three decades. Due to such circumstances, the estimate of species effective population size might help analyze its viability. Single-sample estimator was used to estimate current effective population size (Ne) of broad-snouted caiman populations in representative areas of the species range in Brazil. For the analyzes, genotypes previously obtained were used for subpopulations of the captive colony of the University of São Paulo (USP) and for wild subpopulations. The microsatellites used were Amiμ8, Amiμ11, Amiμ13, Amiμ20, Claμ2, Claμ5, Claμ6, Claμ7, Claμ8, Claμ9 and Claμ10. The 11 loci analyzed produced 18.27 alleles on average. Wild populations showed significant genic and genotypic differentiation among them (p < 0.01). Population structure analyses (Rho-statistics) at Genepop for all loci was 0.376. The effective number of population sizes (Ɵ, the Ne estimator of Migrate-n software) of Caiman latirostris populations from the captive colony at the USP was 117.1 (N = 7) for NRF, 115.2 (N = 4) for Fm1, and 107.5 (N = 4) for Fm5, on the oposite side, the mean value of theta (Ɵ) was 7.7 (N = 51) for the Atlantic SE basin wild population, 9.4 (N = 42) for the Atlantic N/NE basin wild population and 9.6 (N = 91) for the Paraná basin wild population. The mean number of migrants (M of Migrate-n software) varied from 1.7 to 1.9 estimated by migrate-n are comparable to 1.3 estimated by Genepop. It is observed that in most wild population only few adults effectively contribute to genetic variation. The following guidlines are proposed as management actions: (1) an update of population sampling; (2) redefinition, delimitation and conservation of natural habitats; and (3) new conservation genetic studies with special attention to genetic diversity recovery