ABSTRACT
Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs) and single sponge cells (cytospins) and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs) and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques.
Subject(s)
Cell Culture Techniques , Aquatic Fungi/analysis , In Vitro Techniques , Marine Environment , Marine Fauna , Porifera/microbiology , Environmental Microbiology , Methods , MethodsABSTRACT
Sex determination is important for conservation and population studies, particularly for reproduction programs of threatened species and behavioural ecology. Turdus amaurochalinus, Creamy-bellied Thrush, only exhibits sexual dimorphism during the breeding season, when males are considered to show intense yellow bills, and females and immature males show dark brown bills. The objectives of this study were: 1) to determine the sex of individuals using genetic techniques, and 2) to test the hypothesis that sex dimorphism can be detected by morphometry. This study was carried out at Parque Nacional da Restinga de Jurubatiba, a preserved area located on the North coast of Rio de Janeiro State. The birds were captured using ornithological nets, singly marked with metal rings, weighed, measured and had blood samples collected before being released. The sex of 42 T. amaurochalinus individuals was determined using the CHD gene marker. A total of 20 males and 22 females were identified from June to August, with peak capture frequency in June. Turdus amaurochalinus females and males differed significantly in morphometrical measures. The most important traits to distinguish males from females were wing length (Student t-test=4.34, df=40, p=0.0001) and weight (Student t-test=2.08,df=40, p=0.044): females were heavier and had significantly shorter wing length than males. Females and males were correctly classified in 86% and 75% of cases, respectively, using Discriminant Analysis. The molecular analysis was the most secure method for sex determination in the studied species. Rev. Biol. Trop. 59 (2): 789- 794. Epub 2011 June 01.
La determinación del sexo es importante para la conservación y los estudios poblacionales. Turdus amaurochalinus no presenta aparente dimorfismo sexual. El objetivo de este estudio fue determinar el sexo a través de una técnica genética, mediante el uso del marcador del gen CHD y se puso a prueba la hipótesis de que el dimorfismo sexual puede ser detectado por morfometría. Este estudio se llevó a cabo en el Parque Nacional da Restinga de Jurubatiba, una zona protegida situada en la costa norte de Río de Janeiro. Las aves fueron capturadas con redes de niebla, los individuos se marcaron con anillos de metal, se pesaron, medieron y se les tomó una muestra de sangre antes de ser liberados. Un total de 20 machos y 22 hembras fueron identificados en el área de estudio desde junio hasta agosto, con la frecuencia máxima de captura en junio. La prueba de t-student fue usada para evaluar si hembras y machos se diferencian considerablemente en relación a medidas morfométricas. Los rasgos más importantes para distinguir machos de hembras fueron la longitud del ala y el peso: las hembras eran más pesadas y tenían longitud de ala considerablemente más corta que los machos. Hembras y machos fueron correctamente clasificados en un 86% y 75% de casos respectivamente, donde se usó un análisis discriminante. El análisis molecular es el método más seguro para la determinación sexual en la especie estudiada.