Cytogenetic studies on the painted turtle (Trachemys dorbigni: Testudinata, Emydidae)

El objetivo de este trabajo fue caracterizar la estructura cromosómica de Trachemys dorbigni (Testudinata, Emydidae) aportando información a los escasos antecedentes citogenéticos reportados. Se analizaron 14 ejemplares alojados en instalaciones de la Estación Zoológica Experimental Granja La Esmeralda de la ciudad de Santa Fe provenientes de diferentes decomisos, por lo que sus lugares de origen no pudieron ser establecidos. Las preparaciones se obtuvieron a partir de cultivo de sangre entera y el análisis de metafases se realizó mediante tinción convencional Giemsa y tinciones diferenciales C y NOR. Los resultados determinaron un número cromosómico diploide de 2n=50, y los cromosomas se clasificaron según su morfología en 6 pares de metacéntricos/submetacéntricos, 10 pares de acrocéntricos/telocéntricos y 9 pares de microcromosomas; dicha clasificación fue corroborada mediante promedio de relación de brazos e índice centromérico. No se observaron diferencias morfológicas entre cariotipos de machos y hembras, mientras que los bandeos C revelaron heterocromatina en posición telomérica y en bloque y los bandeos NOR determinaron una región organizadora nucleolar en el par 13, en posición terminal en ambos telómeros. Este trabajo permite ampliar los escasos antecedentes citogenéticos en esta especie, debiéndose profundizar el estudio mediante bandeo con fluorescencia para corroborar o complementar los resultados obtenidos.The aim of the present study was to characterize the karyotypic structure of Trachemys dorbigni (Testudinata, Emydidae) contributing information to the scarce cytogenetic data reported. Fourteen specimens housed on the premises of the Estación Zoológica Experimental “Granja La Esmeralda” of Santa Fe City were analyzed. The exact place of origin could not be determined since they came from various confiscations. Preparations were obtained from whole blood culture. The analysis of metaphases was performed by conventional Giemsa staining and differential banding. The results showed a diploid chromosome number of 2n=50. Chromosomes were classified according to their morphology in 6 pairs of metacentric/submetacentric, 10 pairs of acrocentric / telocentric and 9 pairs of microchromosomes; this classification was corroborated by average arm ratio and centromeric index. No morphological differences were observed between the karyotypes of males and females, whereas C banding showed heterochromatin telomeric position, and NOR staining determined a nucleolar organizer region in pair 13, in both telomeres in terminal position. This work allows broadening the limited cytogenetic background regarding this species, being necessary further studies using fluorescence banding to corroborate or supplement our results.Fil: Salas, Anabel. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; ArgentinaFil: Imhof, Alba. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Cs.naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (fhuc-unl/maspyma); ArgentinaFil: Amavet, Patricia Susana. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

DNA extraction from skins of wild (Hydrochoerus hydrochaeris and Pecari tajacu) and domestic (Sus scrofa domestica) species using a novel protocol.

Sometimes, commercial products obtained from wild animals are sold as if they were from domestic animals and vice versa. At this point of the productive chain, legal control of possible wildlife products is difficult. Common in the commerce of northern Argentina, skins of two wild species, the carpincho and the collared peccary, look very similar to each other and to those of the domestic pig; it is extremely difficult to differentiate them after they have been tanned. Because there was no an adequate methodology to discriminate between leather of these three species, we developed a new methodology of DNA extraction from skin and leather. This new method involves digesting a leather sample using proteinase K, followed by precipitation of proteins with 5 M NaCl, cleaning with absolute isopropanol and DNA precipitation with 70% ethanol. DNA is hydrated in Tris-EDTA buffer. This protocol provided good-quality DNA suitable for analysis with molecular markers. This new protocol has potential for use in identifying leather products of these species using molecular markers based on RAPDs.Fil: Ojeda, Guillermo Nicolás. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; ArgentinaFil: Amavet, Patricia Susana. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; ArgentinaFil: Rueda, Eva Carolina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; ArgentinaFil: Siroski, Pablo Ariel. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentin

Estudios genético-poblacionales de Caiman latirostris (Reptilia, Alligatoridae) en Santa Fe, Argentina: una revisión a través del tiempo

Los estudios genéticos en Caiman latirostris (Yacaré overo) en Santa Fe, Argentina, comenzaron con un análisis citogenético en comparación con Caiman yacare. El cariotipo en ambas especies consistió de 42 cromosomas, con un patrón de bandas C difuso y un solo par de cromosomas con NOR. Debido a la falta de diferencias claras entre ambos cariotipos, decidimos usar marcadores moleculares en el análisis de esta especie. Cuatro isoenzimas: Esterasas, Isocitrato Dehidrogenasa, Malato Dehidrogenasa, y Superóxido Dismutasa se analizaron en animales provenientes de cuatro poblaciones santafesinas. En todos los casos se encontraron valores nulos de heterocigosis. Se analizaron 7 primers para RAPD y sólo 13.73 % de 233 marcadores analizados resultaron polimórficos. Los resultados para polimorfismos, heterocigosis y número medio de alelos por locus en las poblaciones fueron niveles bajos a intermedios. El análisis de AMOVA indicó que casi toda la variación existe dentro de las poblaciones, significando que varios alelos son compartidos entre las poblaciones. Debido a que los marcadores RAPD pueden ser menos eficaces en detectar variaciones que los microsatélites en poblaciones de cocodrilianos, en estos momentos estamos comenzando a utilizar esta técnica en el análisis poblacional. Hasta el momento se han amplificado positivamente cuatro primers, y detectamos indicios de la existencia de más de un padre en tres familias. Los datos genéticos de especies nativas son fundamentales para establecer y evaluar planes de manejo, y el análisis de variabilidad es básico para el conocimiento biológico de las especies, aportando a la sistemática, la ecología y la biodiversidad.Fil: Amavet, Patricia Susana. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rosso, Daniel Esteban. Universidad Nacional del Litoral; ArgentinaFil: Markariani, Rosa. Universidad Nacional del Litoral; ArgentinaFil: Larriera, Alejandro. Universidad Nacional del Litoral; Argentin

Back Home: Strategies for the Reintroduction of Chelonoidis chilensis Into the Wild

The Argentinian Tortoise (Chelonoidis chilensis) is included in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora and classified as Vulnerable by the International Union for the Conservation of Nature; nevertheless, they are illegally traded as pets in Argentina. Many tortoise owners who kept them as pets gave their tortoises to Protectora Rosario, which sent them to a Wildlife Rescue Center in Santa Fe, Argentina. Our goals were to clinically and behaviorally evaluate these tortoises to analyze the feasibility of their release; determine the genetic population of origin of each specimen by identifying its mitochondrial haplotype; and generate educational actions to prevent the commercialization of this species. The genetic analysis identified six haplotypes, four corresponded to the Dry Chaco region, one with the Monte region, and two were new haplotypes for the species. Clinical and behavioral evaluations after a quarantine process showed that the tortoises were suitable to be reintroduced into the wild because they were disease-free, they showed a correct usage of their microhabitat, reacted to environmental stimuli, and had independence from humans for their food. Monitoring showed that the reintroduction of the tortoises was successful: we relocated 23 of the 28 released tortoises during the 6 mo after their release and none showed signs of disease or damage. We suggest that the evaluation methods and genetic analysis we applied in this study can be used as a baseline for the establishment of standardized protocols to apply in reintroduction programs for other tortoises.Fil: Amavet, Patricia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; ArgentinaFil: Sciabarrasi Bagilet, Antonio Alejandro. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de Ambiente.; ArgentinaFil: Sánchez, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; Argentina. Universidad Nacional de Tierra del Fuego, Antártida e Islas del Atlántico Sur. Instituto de Ciencias Polares, Ambientales y Recursos Naturales; ArgentinaFil: Bersezio, Natalia Soledad. Gobierno de la Provincia de Santa Fe. Ministerio de Ambiente.; ArgentinaFil: Castiglioni, Mariela. Gobierno de la Provincia de Santa Fe. Ministerio de Ambiente.; ArgentinaFil: Siroski, Pablo Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentin

As ferramentas moleculares aplicadas na conservação dos crocodilianos

Nos últimos tempos, estudos de genética cresceram rapidamente graças ao avanço da tecnologia e das ferramentas moleculares, e os resultados obtidos nas análises genéticas se tornaram importantes ferramentas para a conservação e manejo, permitindo aprofundamento sobre a estrutura populacional, padrões de dispersão, fluxo gênico e hibridização interespecífica, dentre outros parâmetros. Os crocodilianos são um dos grupos de vertebrados mais fascinantes. Além de suas características intrínsecas (i.e., longevidade), representam um grupo bem-sucedido de organismos que sofreram poucas alterações ao longo de milhões de anos, sendo estes animais importante fonte de informação para quem pesquisa os avanços do ponto de vista molecular. A capacidade de resistir ao ataque de microrganismos poderia ser um dos motivos de seu sucesso e longevidade e, por isso, a descoberta dos componentes moleculares envolvidos permitiria um conhecimento aprofundado desses fatos. Neste aspecto, o entendimento dos processos evolutivos frequentemente requer a análise de marcadores polimórficos. Além disso, o impacto biológico resultante da interação com componentes sensíveis à contaminação em diferentes espécies de jacarés pode se manifestar nos níveis genético, imunológico e bioquímico, exercendo um efeito marcante de longo prazo nas populações. Marcadores moleculares, imunológicos e bioquímicos constituem uma bateria de ferramentas complementares altamente explicativas na avaliação dos efeitos nos crocodilianos causados pelo uso massivo dos pesticidas, permitindo obter mais informações sobre o significado biológico específico e o impacto real das alterações produzidas por defensivos agrícolas, gerando ferramentas preditivas para a proteção ambiental dessas e de outras espécies silvestres de nossa região. Especificamente, em toda a sua área de distribuição, um grande número de espécimes de C. latirostris foram caçados ilegalmente, razão pela qual foi necessário estabelecer diferentes estratégias para evitar sua extinção. Em conclusão, os recursos genéticos têm permitido abordar o estudo dos crocodilianos do ponto de vista populacional, ecológico, ecossistêmico e produtivo.Lately, genetics studies advanced quickly using molecular tools and the technology. Besides their intrinsic characteristics like longevity, habitats, and some other interesting aspects, crocodilians are one of the most fascinating groups within vertebrates. They represent a successful group of organisms that have suffered little alteration over millions of years, thus these animals are an important source of information for those researching advances from a molecular point of view. The ability to resist the attack of microorganisms could be one of the reasons for their success and longevity so the discovery of the molecular components involved would allow a thorough understanding of these facts. In addition, the information acquired from genetic analyses is important for conservation and management and allow to depth on population structure, patterns of dispersal and gene flow, interspecific hybridization, among other parameters, as to understand evolutionary processes require the analysis of polymorphic markers. Also, the biological impact resulting from the interaction with contamination-sensitive components in caiman species can be manifested at the genetic, immunological, and biochemical levels, exerting a marked long-term effect on populations. Genetic, immunological and biochemical molecular markers constitute a battery of complementary tools that are highly explanatory in the evaluation of the effects caused by mass use pesticides on crocodilians and will allow us to obtain more information regarding the specific biological significance and real impact of the alterations produced by pesticides on these species, providing predictive tools for the environmental protection of these and other wild species in our region.Fil: Siroski, Pablo Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Amavet, Patricia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Bassetti, Luís Antonio Bochetti. Universidade de Sao Paulo; Brasi

Distribution studies of the genetic variability and analysis of mating system in wild populations of Caiman latirostris (Broad-snouted caiman) (Reptilia, alligatoridae)

Caiman latirostris (yacaré overo) es una de las dos especies de cocodrilianos citadas para nuestro país. Sus poblaciones silvestres son objeto de manejo en Argentina, mediante programas que utilizan el sistema de ranching (cosecha de huevos silvestres para cría en granjas), para su uso sustentable y la conservación de su hábitat. C. latirostris, en nuestro país, se encuentra en el Apéndice II de CITES, el cual permite el comercio regulado de su cuero y carne, y se ha convertido en una especie de gran importancia comercial, a nivel nacional e internacional. Esta tesis incluye estudios genético-poblacionales del yacaré overo en la provincia de Santa Fe, Argentina. Se realizaron análisis de variabilidad, diferenciación y estructura genética empleando isoenzimas, marcadores RAPD y caracteres morfométricos, y un estudio de paternidad utilizando marcadores microsatélites. Los resultados obtenidos indican que las poblaciones analizadas de yacaré overo poseen valores de variabilidad genética de bajos a intermedios, diferenciación poblacional significativa y variabilidad fenotípica alta para algunos de los rasgos morfométricos estudiados. Además, se hallaron indicios de que el sistema de apareamiento de C. latirostris podría incluir el comportamiento de múltiple paternidad, al detectar más de un progenitor paterno en al menos una de las familias analizadas.Caiman latirostris (broad-snouted caiman) is one of the two crocodilian species cited for Argentina. Their wild populations are subject to management in our country, by means of programs that use ranching system (harvest of wild eggs, captive rearing and reintroduction to wild) for their sustainable use and habitat conservation. In Argentina, C. latirostris is included in the Appendix II of CITES, therefore, the regulated trade of meat and leather is allowed, and this species has turned into an important resource at both national and international levels. This thesis includes genetic population studies about broad-snouted caiman in Santa Fe province, Argentina. Analyses related to variability, differentiation and genetic structure were carried out through isozyme electrophoresis, RAPD markers, and quantitative traits. On the other hand, paternity studies were conducted using microsatellite markers. The obtained results indicate that analyzed broad-snouted caiman populations have low to intermediate genetic variability values, significant population differentiation, and high phenotypic variability for some of the studied morphometric traits. In addition, we found indications that C. latirostris mating system could include multiple paternity behavior, since we found more than one paternal progenitor in at least one of the analyzed families.Fil:Amavet, Patricia Susana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Baseline values of immunologic parameters in the lizard Salvator merianae (Teiidae, Squamata)

The genus Salvator is widely distributed throughout South America. In Argentina, the species most abundant widely distributed is Salvator merianae. Particularly in Santa Fe province, the area occupied by populations of these lizards overlaps with areas where agriculture was extended. With the aim of established baseline values for four immunologic biomarkers widely used, 36 tegu lizards were evaluated tacking into account different age classes and both sexes. Total leukocyte counts were not different between age classes. Of the leucocytes count, eosinophils levels were higher in neonates compared with juvenile and adults; nevertheless, the heterophils group was the most prevalent leukocyte in the peripheral blood in all age classes. Lymphocytes, monocytes, heterophils, azurophils and basophils levels did not differ with age. Natural antibodies titres were higher in the adults compared with neonates and juveniles lizards. Lastly, complement system activity was low in neonates compared with juveniles and adults. Statistical analysis within each age group showed that gender was not a factor in the outcomes. Based on the results, we concluded that S. merianae demonstrated age (but not gender) related differences in the immune parameters analyzed. Having established baseline values for these four widely-used immunologic biomarkers, ongoing studies will seek to optimize the use of the S. merianae model in future research.Keywords: Biomarkers, Immune system, Reptilian, Salvator merianae, Sentinel mode

Characterization of genetic diversity of Trachemys dorbigni and Phrynops hilarii

The utilization of RAPD and ISSR molecular markers is proposed to initiate studies of genetic variability in Phrynops hilarii and Trachemys dorbigni, two species of fresh water turtles distributed in South America. Three primers of RAPD and four of ISSR were selected and the amplified products of these markers were evaluated by electrophoretic runs in agarose and polyacrylamide gels. The levels of heterozygosity, Shannon index and different allele numbers were slightly higher in P. hilarii for both types of markers. Levels of polymorphism were also higher in P. hilarii than T. dorbigni and both were elevated compared to those recorded for other species. The fact that similar results were obtained with both types of markers for all estimates of diversity highlights the usefulness and validity of the RAPD technique. The molecular markers used were found potentially useful for analysing future temporal and spatial distribution of genetic diversity in both species, expanding scales work.Fil: Guidetti, Brenda Yamile. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Siroski, Pablo Ariel. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Cs.naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (fhuc-unl/maspyma); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Amavet, Patricia Susana. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

Analysis of morphological variability and heritability in the head of the Argentine Black and White Tegu (Salvator merianae): undisturbed vs. disturbed environments

The heterogeneity of biotic and abiotic factors influencing fitness produce selective pressures that promote local adaptation and divergence among different populations of the same species. In order for adaptations to be maintained through evolutionary time, heritable genetic variation controlling the expression of the morphological features under selection is necessary. Here we compare morphological shape variability and size of the cephalic region of Salvator merianae specimens from undisturbed environments to those of individuals from disturbed environments, and estimated heritability for shape and size using geometric morphometric and quantitative genetics tools. The results of these analyzes indicated that there are statistically significant differences in shape and size between populations from the two environments. Possibly, one of the main determinants of cephalic shape and size is adaptation to the characteristics of the environment and to the trophic niche. Individuals from disturbed environments have a cephalic region with less shape variation and also have a larger centroid size when compared to individuals from undisturbed environments. The high heritability values obtained for shape and size in dorsal view and right side view indicate that these phenotypic characters have a great capacity to respond to the selection pressures to which they are subjected. Data obtained here could be used as an important tool when establishing guidelines for plans for the sustainable use and conservation of S. merianae and other species living in disturbed areas.Fil: Imhoff, Carolina Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales; ArgentinaFil: Giri, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; ArgentinaFil: Siroski, Pablo Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Amavet, Patricia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentin