The multigenerational effects of water contamination and endocrine disrupting chemicals on the fitness of Drosophila melanogaster.

Water pollution due to human activities produces sedimentation, excessive nutrients, and toxic chemicals, and this, in turn, has an effect on the normal endocrine functioning of living beings. Overall, water pollution may affect some components of the fitness of organisms (e.g., developmental time and fertility). Some toxic compounds found in polluted waters are known as endocrine disruptors (ED), and among these are nonhalogenated phenolic chemicals such as bisphenol A and nonylphenol. To evaluate the effect of nonhalogenated phenolic chemicals on the endocrine system, we subjected two generations (F0 and F1) of Drosophila melanogaster to different concentrations of ED. Specifically, treatments involved wastewater, which had the highest level of ED (bisphenol A and nonylphenol) and treated wastewater from a constructed Heliconia psittacorum wetland with horizontal subsurface water flow (He); the treated wastewater was the treatment with the lowest level of ED. We evaluated the development time from egg to pupa and from pupa to adult as well as fertility. The results show that for individuals exposed to treated wastewater, the developmental time from egg to pupae was shorter in individuals of the F1 generation than in the F0 generation. Additionally, the time from pupae to adult was longer for flies growing in the H. psittacorum treated wastewater. Furthermore, fertility was lower in the F1 generation than in the F0 generation. Although different concentrations of bisphenol A and nonylphenol had no significant effect on the components of fitness of D. melanogaster (developmental time and fertility), there was a trend across generations, likely as a result of selection imposed on the flies. It is possible that the flies developed different strategies to avoid the effects of the various environmental stressors.

A likelihood inference of historical biogeography in the world's most diverse terrestrial vertebrate genus: diversification of direct-developing frogs (Craugastoridae: Pristimantis) across the Neotropics.

The geology of the northern Andean region has driven the evolutionary history of Neotropical fauna through the creation of barriers and connections that have resulted in speciation and dispersal events, respectively. One of the most conspicuous groups of anuran fauna in the Andes and surrounding areas is the direct-developing species of the genus Pristimantis. We investigated the molecular phylogenetic placement of 12 species from the montane Andes of Colombia in a broader geographical context with a new genus-level phylogeny in order to identify the role of Andean orogeny over the last 40million years and the effect of elevational differences in diversification of Pristimantis. We examined the biogeographic history of the genus using ancestral range reconstruction by biogeographic regions and elevational ranges. We recognized the middle elevational band (between 1000 and 3000m) in the Northwestern Andes region of Colombia and Ecuador as a focal point for the origin and radiation of Pristimantis species. Additionally, we found several Andean migrations toward new habitats in Central Andes and Merida Andes for some species groups. We suggest that the paleogeological changes in the Northwestern Andes were the main promoter of speciation in Pristimantis, and may have served as a corridor for the dispersion of lowland species.

DIET OF DIRECT-DEVELOPING FROGS (ANURA: CRAUGASTORIDAE: Pristimantis) FROM THE ANDES OF WESTERN COLOMBIA / Dieta de ranas de desarrollo directo (Anura: Craugastoridae: Pristimantis) de los Andes occidentales de Colombia

We studied the diet of 15 montane frog species of the genus Pristimantis (Craugastoridae) from the Andes of Western Colombia to determine the diet range, breadth of niche and overlap among species. We identified 499 prey items from stomach and intestinal contents of 154 specimens. Prey items were included in 74 different categories. The most common 15 prey categories accounted for 65 % of all frogs' diet. The invertebrate families Isotomidae, Chironomidae, Formicidae, and Tipulidae were the most abundant categories and accounted for 32 % of the frogs' diet. Ten of the 15 frog species were found with at least one item of Araneae. Coleoptera and Tipulidae were found in nine frog species, and Acari and Carabidae in eight frog species. In general, beetles were found in gastrointestinal tracts of all species examined, except for P. quantus, but interpretation needs caution because only one individual of this species was caught. Pristimantis hectus showed a specialized diet, consuming mainly dipterans of the family Chironomidae, while the remaining species showed a generalist diet. Pristimantis palmeri showed niche overlap with P. erythropleura (Фjk = 0.69), P. myops (Фjk = 0.64), and P. orpacobates (Фjk = 0.64). Our results suggest that most of the frogs species studied are generalist, foraging opportunistically on dipterans, arachnids, collembolans, coleopterans, and hymenopterans. Here, we report the diet of montane Pristimantis species and discuss the results in comparison with data on related species in montane and lowland regions.

Estudiamos la dieta de 15 especies de ranas montanas del género Pristimantis (Craugastoridae) en la Cordillera Occidental de Colombia para determinar su variación, amplitud y traslape de nicho. En los contenidos estomacales e intestinales de 154 especímenes se identificaron 499 presas que fueron organizadas en 74 categorías diferentes. Las 15 categorías de presas más comunes representaron el 65 % de la dieta. Las familias de invertebrados Isotomidae, Chironomidae, Formicidae y Tipulidae fueron las más abundantes, representando el 32 % de la dieta de las ranas. Diez de las 15 especies de ranas presentaron al menos un ítem de Araneae. Coleoptera y Tipulidae fueron otras categorías frecuentemente encontradas en los tractos digestivos de las ranas (nueve especies de ranas presentaron estos ítems), seguidas de Acari y Carabidae (ocho especies). En general, los escarabajos fueron encontrados en los tractos gastrointestinales de todas las especies, excepto en P. quantus, pero este resultado debe tratarse con precaución porque solo un individuo de esta especie fue capturado. Pristimantis hectus mostró una dieta especializada consumiendo principalmente dípteros de la familia Chironomidae, mientras que las especies restantes mostraron una dieta generalista. Pristimantis palmeri presentó superposiciones de nicho con P. erythropleura (Фjk = 0,69), P. myops (Фjk = 0,64), y P. orpacobates (Фjk = 0,64). Nuestros resultados sugieren que la mayoría de las especies estudiadas son generalistas que se alimentan de manera oportunista de dípteros, arácnidos, colémbolos, coleópteros e himenópteros. Aquí, reportamos la dieta de especies montanas de Pristimantis y discutimos los resultados con datos de otros estudios en especies relacionadas de regiones montanas y tierras bajas.

Análisis genétioo de la población de gatos del norte y sur de cali, Colombia / Genetic Analysis of the Cat Population of North and South of Cali, Colombia

Los marcadores genéticos del pelaje y malformaciones óseas han permitido caracterizar el perfil genético de más de 400 poblaciones del gato doméstico alrededor del mundo. Hace 15 años se estableció dicho perfil en la ciudad de Cali (Colombia). En este estudio se determinó si el norte y sur de Cali se comportan como subpoblaciones y se comparó el perfil total con el estudio pasado. Se encontró una disminución de la frecuencia alélica de a (no-agouti) y d (dilution), pero un aumento en cinco, especialmente en l (longhair) y c s (siamese). Dichas diferencias pueden atribuirse a la selección humana de características más atractivas y por el flujo génico resultante del crecimiento demográfico de la ciudad, lo que explicaría también el primer reporte de los alelos inhibitor y ticked abyssinian. Se evaluó el equilibrio Hardy-Weinberg para el norte, sur y las dos zonas juntas, usando los loci white spotting y orange, encontrándose desequilibrio en este último para las tres zonas evaluadas debido a un déficit de heterocigotos. Norte y sur se dividieron en dos, y cada sub-muestra presentó equilibrio Hardy-Weinberg, aunque las diferencias en las frecuencias alélicas y heterocigosidades resaltaron microestructura geográfica y una relación entre tiempo de fundación del barrio y heterocigosidad. Norte y sur resultaron ser una población y no subpoblaciones (F ST= 0,0004, D= 0,0017), al igual que las nueve poblaciones colombianas con las que se comparó la presente ciudad. Se sugiere realizar un análisis microgeográfico de flujo génico y la definición de posibles colonias de gatos en Cali.

The coat genetic markers and skeleton abnormalities have allowed characterize the profile from more than 400 domestic cat populations around the world. 15 years ago, that profile was established in the city of Cali (Colombia). In this study it was determined if north and south of the city are subpopulations and it was compared the total profile against past study. A decrease in allele frequency of a (non-agouti) and d (dilution) was found, but an increase of five alleles was found, especially in l (long hair) and c s (siamese). These differences could be attributed to human selection of more attractive characteristics and gene flow resulting from demographic growth city, which would also explain the first report of inhibitor and ticked abyssinian alleles. Hardy-Weinberg equilibrium was evaluated for the north, south and both areas together, using white spotting and orange loci, determining disequilibrium in orange for the three evaluated areas due to a heterozygotes deficit. North and south were divided into two, each sub-sample showed Hardy-Weinberg equilibrium, although allele frequencies and heterozygosities highlighted microgeographic structure and a relationship between founding time of the neighborhood and heterozygosity. North and south are a single population and aren´t subpopulations (F ST= 0,0004, D= 0,0017), as well as nine Colombian populations with which this city was compared. It is suggested to make a microgeographical gene flow analysis and the definition of possible cat colonies in Cali.

ECOLOGÍA ALIMENTARIA DE LA RANA DE LLUVIA ENDÉMICA Pristimantis jubatus (CRAUGASTORIDAE) EN EL PARQUE NACIONAL NATURAL MUNCHIQUE, COLOMBIA / Feeding Ecology of the Endemic Rain Frog Pristimantis jubatus (Craugastoridae) in Munchique National Park, Colombia

Se estudió la dieta de la rana de lluvia Pristimantis jubatus con base en los contenidos estomacales e intestinales de 29 individuos colectados en tres diferentes localidades del Parque Nacional Natural Munchique, Colombia. La dieta estuvo compuesta exclusivamente de artrópodos, identificándose un total de 66 presas correspondientes a 32 taxa. Diptera e Hymenoptera fueron los artrópodos más frecuentemente encontrados, 19,7 % y 16,7 % respectivamente, seguidos de Araneae (15,1 %) y de Coleoptera (12,1 %). Esta especie no mostró preferencia por ninguno de los tipos de presa y presentó un alto valor de amplitud de nicho (0,68), lo cual indica una dieta generalista. Adultos y juveniles forrajean en diferentes alturas de percha, encontrándose los primeros a alturas mayores que los segundos. El número de categorías encontradas y el uso del hábitat muestran a P. jubatus como una especie que puede consumir oportunamente cualquier artrópodo disponible en los estratos del bosque donde forrajea.

We studied the diet of the rain frog Pristimantis jubatus based on stomach and intestinal contents of 29 individuals from three different localities at Munchique National Park, Colombia. The diet was composed exclusively of arthropods, comprising a total of 66 prey which were in 32 taxa. The most frequent dietary items were Diptera and Hymenoptera, 19.7 % and 16.7 % respectively, followed by Araneae (15.1 %) and Coleoptera (12.1 %). Plant consumption was considered incidental because of its low frequency. This species showed no preference for any type of prey, with a high value of niche breadth (0.68) indicating a generalist diet. Adults and juveniles occupy different perch height. Habitat use and the variety of items found show P. jubatus as a species that can consume any kind of arthropod available in the strata of the forest where it is foraging.

Genetic variability of five Periplaneta americana L. (Dyctioptera: Blattidae) populations in southwestern Colombia using the AFLP molecular marker technique.

The genetic structure of five Periplaneta americana (L.) populations from three cities (Cali, Popayán and Buenaventura) located in southwestern Colombia was estimated using the AFLP molecular marker technique. A set of 174 loci were analyzed, 120 of which were polymorphic. Genetic diversity was estimated at different geographic levels for the total population and between cities. Genetic diversity averaged 0.32. The largest gene flow was detected between two population from Cali (F ST=0.088; Nm=2.6) and the lowest from the third population in this city (Univalle) and Popayán and Buenaventura (F ST=0.13; Nm=1.6 and F ST=0.12; Nm=1.8, respectively). The individuals' low mobility, the population' geographic separation, and possibly genetic drift have resulted in the population of this cockroach species having a significant degree of structuring, both between cities (Phist=0.13; P<0.001) and within them (Phist=0.87; P<0.001). These results suggest a high degree of subdivision within the P. americana population studied.

DIVERSIDAD Y ESTRUCTURA GENÉTICA DE TRESPOBLACIONES AFRODESCENDIENTES DEL SUROCCIDENTECOLOMBIANO A PARTIR DE 8 STR’S. / Genetic Structure and Diversity of Three Colombian Southwest Afrodescendent Populations Using 8 STR’s.

Con el objetivo de estimar la diversidad, la estructura y el flujo génico de tres poblaciones afrodescendientes del suroccidente colombiano (Buenaventura, Mulaló y Tumaco), se analizaron los alelos revelados por ocho STR’s autosómicos en 78 indi-viduos no relacionados, mediante amplificación por PCR y comparación con escaleras alélicas de cada sistema corridas en geles de poliacrilamida al 8%. Los resultados se compararon con las poblaciones amerindias Awa-Kuaikier y Coyaima, y las mestizas del Valle del Cauca y de Cauca. Se encontró que las muestras afrodescendientes y amerindias fueron moderadamente diversas (h entre 0,768±0,414 y 0,796±0,424), mientras que las mestizas mostraron índices mayores (>0,803), lo que puede ser consecuencia del mestizaje con amerindios, el cual puede explicar la alta endogamia observada para éstas. El AMOVA exhibió estructuración moderada entre las poblaciones afrodescendientes (FST= 0,098; p<0,05), y alta entre los tres grupos étnicos comparados (FST=0,26723; p<0,05). Las distancias genéticas favorecieron la cercanía entre Tumaco y Buenaventura, soportada por la tasa de migración encontrada (34,298), al igual que al interior de las poblaciones amerindias y mestizas. Las diferencias observadas entre Mulaló y las otras poblaciones negras quizá se expliquen porque es un aislado poblacional más cerrado. El árbol NJ mostró la relación más cercana entre las poblaciones amerindias y mestizas, además del carácter ancestral de las afrodescendientes. Esto sustenta la idea del flujo genético mantenido entre las tres etnias, principalmente entre las poblaciones amerindias y mestizas analizadas, soportado por las distancias genéticas, las tasas de migración y la matrilinealidad amerindia reportada en la literatura.

To estimate the diversity, structure and genetic flow in three colombian southwest afrodescendent populations (Buenaventura, Mulaló y Tumaco), the alleles revealed by 8 autosomal STR’s were analyzed in 78 no related individuals, by the use of PCR and comparison with specific allelic ladders for every system resolved by polyacrylamide gel (8%). The results were compared with 2 amerindian populations (Awa-Kuaikier and Coyaima) and 2 mixed colombian populations (Valle del Cauca and Cauca). For the afrodescendent and amerindian populations was found moderate diversity (h between 0.768±0.414 and 0.796±0.424), in contrast, the mixed population showed higher rates (>0.803), which is probably caused by mixing with amerindians, that also can explain the high endogamy seen in mixed populations. The AMOVA exhibited moderate genetic structure between the afrodescendent populations (FST= 0.098; p<0.05), but higher between the three ethnical groups compared (FST=0.26723; p<0.05). The closer genetics distances are in favor of Tumaco and Buenaventura, supported for the migration rate found (34.298), which was the same inside of amerindian and mixed populations. Maybe, because Mulaló is a closed isolated population, its differences in front others afrodescendent populations are explained. The neighbor-joining tree showed nearest relations among amerindian and mixed populations, furthermore, the ancestral character for the afrodescendents. That sustains the idea of genetic flow maintained between the 3 ethnical groups, principally between amerindian and mixed populations, supported because the genetic differences, migration rates and amerindian matrilineality reported in the literature.

Genetic variability of Five Periplaneta americana L. (Dyctioptera: Blattidae) populations in southwestern Colombia using the AFLP molecular marker technique

The genetic structure of five Periplaneta americana (L.) populations from three cities (Cali, Popay¨¢n and Buenaventura) located in southwestern Colombia was estimated using the AFLP molecular marker technique. A set of 174 loci were analyzed, 120 of which were polymorphic. Genetic diversity was estimated at different geographic levels for the total population and between cities. Genetic diversity averaged 0.32. The largest gene flow was detected between two population from Cali (F ST = 0.088; Nm = 2.6) and the lowest from the third population in this city (Univalle) and Popay¨¢n and Buenaventura (F ST = 0.13; Nm = 1.6 and F ST = 0.12; Nm = 1.8, respectively). The individuals' low mobility, the population' geographic separation, and possibly genetic drift have resulted in the population of this cockroach species having a significant degree of structuring, both between cities (¦µst = 0.13; P < 0.001) and within them (¦µst = 0.87; P < 0.001). These results suggest a high degree of subdivision within the P. americana population studied.

EVALUACIÓN DE DOS MEDIOS DE CULTIVO Y HEREDABILIDAD DE PRODUCTIVIDAD Y TIEMPO DE DESARROLLO PARA TRES MUTANTES DE Drosophila melanogaster (DROSOPHILIDAE) / Evaluation of Two Culture Media and Heritability of Productivity and Development Time for Three Mutants of Drosophila melanogaster (Drosophilidae)

Con el objetivo de investigar el efecto del medio de cultivo en la productividad y tiempo de desarrollo huevo-adulto de una cepa silvestre y tres cepas mutantes (CyLv, vg, w) de Drosophila melanogaster, se examinaron dos tipos de medios: banano y naranja. Para esto se empleó un diseño con dos factores, medio de cultivo y tipo de cepa, para un total de ocho tratamientos con cinco repeticiones cada uno. Se obtuvo que la productividad y el tiempo de desarrollo dependen del medio de cultivo y el tipo de cepa, encontrándose mayor productividad en el medio de naranja. La cepa silvestre presentó la mayor productividad y el menor tiempo de desarrollo en los dos medios (α=0,05). El análisis genético evidenció una heredabilidad baja y una variación fenotípica debida en su mayor parte al componente de interacción genotipo-ambiente, lo que explica la diferencia en el patrón de productividad y tiempo de desarrollo entre medios de cultivo.

With the objective of investigating the effect of the culture media in the productivity and development time egg-adult of Drosophila melanogaster in +/+ and three mutants (CyLv, vg, w), two culture media: banana and orange, were evaluated. An experimental design with two factors: culture media and kind of flies-stock, were tested, for a total of eight treatments with five replicas each one. The productivity and development time depend on culture media and kind of flies-stock, and the biggest productivity was in the orange culture media. The +/+ presented the biggest productivity and lowest development time in both culture media (α=0.05). The genetic analysis showed a low heritability and the phenotypic variation was due, in a mayor part, to the component of the interaction genotype-environment that explains the difference in the patron of productivity and development time between culture media.

Caracterización molecular de tres líneas de Cavia porcellus mediante la aplicación de AFLP / Molecular characterization of three populations of Cavia porcellus with AFLP markers

Los marcadores moleculares son una herramienta eficaz para determinar variabilidad genética entre y dentro de poblaciones, pero en el caso de Cavia porcellus, no existen reportes referentes al uso de estas técnicas. Con los marcadores moleculares AFLP´s (Amplified Fragment Length Polimorphism), se analizaron tres poblaciones, dos criollas y una mejorada genéticamente, sometida a selección durante varias generaciones y obtenida a partir del cruzamiento entre animales peruanos y nativos de Nariño. Para obtener los marcadores moleculares AFLP´s (Amplified Fragment Lenght Polimorphism), se utilizaron en total cinco combinaciones de cebadores, tres combinaciones recomendadas para el orden Rodenthia y dos por la casa fabricante del Kit, de las cuales sólo una de ellas, con 116 loci, permitió establecer diferencias entre las poblaciones estudiadas, de acuerdo con el valor de distancia genética insesgada de Nei (p<0.01). Las dos poblaciones criollas constituyeron un grupo estrechamente relacionado y distante de la población mejorada genéticamente, lo que indica que los animales importados absorbieron al criollo. De acuerdo con los valores de heterocigosidad promedio, que variaron entre 0.48% y 14.48%, y el porcentaje de polimorfismo que osciló entre 0.00% y 39.65%, se deduce una baja variabilidad intrapoblacional, siendo la población mejorada genéticamente la más polimórfica. La baja variabilidad entre los animales mejorados, se explica por la intensa selección a la que han sido sometidos, mientras que en los núcleos criollos este fenómeno puede atribuirse al bajo tamaño efectivo en las dos poblaciones. Los resultados de esta investigación sugieren un replanteamiento de los programas de mejoramiento genético y conservación de los recursos genéticos autóctonos en la región.

Molecular markers are a powerful tool to determine genetic variability within and among populations, but for the Cavia porcellus there are no reports on the use of these techniques. Three populations, two native and another one, genetically improved which was obtained by crossing native and Peruvian animals and submitted to genetic selection through several generations were analyzed by means of AFLP markers. Five primer’s combinations recommended for Rodenthia were used, but only one allowed to establish significant differences (p<0.01) according to unbiased Nei´s distance Value. Both native populations were grouped in a cluster genetically distant from the genetically improved animals. This showed that foreign animals absorbed the native populations. The average heterosigosity between 0.48% and 14.48% and the percentage of polymorphisms between 0.00% and 39.65% allow to conclude that there was a low variability between the populations, but the population genetically improved was the most polymorphic. The low variability within the improved animals it can be explained because of the intensive selection procedures use with them, whereas within the native populations can be explained because of their very low populations effective size. These results suggest that there is a need to restate the genetic improvement and preservation programs of the native Cavia porcellus in the southwest region of Colombia.

Polimorfismos en la longitud de fragmentos amplificados (AFLP´s) a partir de muestras de sangre almacenadas en tarjetas FTA®para la especie Cavia porcellus Lin. (Rodentia: Caviidae). / Amplified Fragments Length Polymorphisms (AFLP´s) from blood samples stored in FTA® cards for Cavia porcellus

Una manera eficaz de establecer el grado de variabilidad entre y dentro de poblaciones, es a través del análisis de polimorfismos de ADN con marcadores moleculares como los AFLP`s. En este artículo se presenta una metodología que combina la utilización de tarjetas de FTA® (Whatman Bioscience, Cambridge) para colección y conservación de muestras de sangre, con los procedimientos de extracción de ADN y obtención de marcadores AFLP´s, aspectos sobre los cuales no existen antecedentes para la especie Cavia porcellus. Se utilizaron muestras de ADN procedentes de tres poblaciones, dos criollas y una mejorada genéticamente obtenida a partir de un pie de cría procedente del Perú y sometida a selección en Colombia durante varias generaciones. Todos los animales procedieron de la Granja “Botana”, propiedad de la Universidad de Nariño, Pasto-Colombia. Para la detección de polimorfismos en la longitud de los fragmentos (AFLP`s) se utilizaron uno, tres y cinco discos FTA® de 1.2 mm, cada disco con aproximadamente 25 ng de ADN. Los ensayos indicaron que los mejores productos de amplificación, para la visualización de AFLP´s, se obtuvieron de muestras con tres discos de FTA por individuo, lo que sugiere que con esta metodología,75 ng de ADN por animal son suficientes para detectar polimorfismos de alta calidad en el genoma de Cavia porcellus. Se recomienda el uso de las tarjetas de FTA para el estudio genético de poblaciones de Cavia porcellus, con las modificaciones metodológicas descritas en este artículo para marcadores AFLP´s.

A methodology that includes the use of FTA® (Whatman Bioscience, Cambridge) to collect and store animals` blood samples and the procedures to extract and to get AFLP markers is presented in this paper. A review of the literature indicates that there are no reports concerning both aspects for the Cavia porcellus case. To reach our goal blood samples of three populations – Two native ones and other genetically improved- were obtained through heart puncture. This blood was stored in the FTA cards in order to extract, purify, amplify and analyze their DNA forms. All of the animals came from “Botana” farm of the Universidad de Nariño, located in Pasto, Colombia. For amplifying the AFLP one, three and five 1.2 mm FTA disks of approximately 75 ng of DNA per disk where used. The tests indicated that the best products to amplify and to visualize the AFLP where those ones obtained from samples of three FTA disks per animal. This suggests that 75 ng of DNA per animal is enough to generate AFLP of high quality in the Cavia porcellus` genome. We recommend the use of FTA cards to carry out genetic analyses in the Cavia porcellus, including the methodology modifications presented in this paper.