Las secreciones de la piel de Agalychnis spurrelli han probado tener una marcada actividad antimicrobiana sobre diferentes microrganismos patógenos por la presencia de biomoléculas en ellas. Se realizaron análisis moleculares del ARN mensajero de la piel de A. spurrelli para determinar el tipo de péptido antimicrobiano presente en las secreciones cutáneas de esta especie. Para amplificar las secuencias precursoras de los péptidos maduros, se utilizaron iniciadores específicos que contienen secuencias altamente conservadas. Como resultado se obtuvo una secuencia de ADN complementario de 357 pb, la cual fue comparada con sus ortólogos de otras especies de la misma subfamilia Phyllomedusinae, se lograron índices de identidad muy altos para precursores de dermaseptinas. Finalmente, para el análisis de la secuencia de ADN complementario, se tradujo la secuencia nucleotídica por codones, con lo que se obtuvo una secuencia aminoacídica. Dicha secuencia posee las características particulares de péptidos de la familia de las dermaseptinas: una secuencia altamente conservada, una propieza acídica con los aminoácidos Lisina y Arginina y el extremo C-terminal variable. En conclusión, la secreción total de Agalychnis spurrelli contiene un péptido de la familia de las dermaseptinas o un péptido relacionado con ellas.

Spiralia represents one of the main clades of bilaterally symmetrical metazoans (Bilateria). This group is of particular interest due to the remarkable conservation of its early developmental pattern despite of the high diversity of larval and adult body plans. Variations during embryogenesis are considered powerful tools to determine ancestral and derived characters under a phylogenetic framework. By direct observation of embryos cultured in vitro, we analyzed the early cleavage of the euopisthobranchs Aplysia cf. californica. We used tubulin immunocytochemistry to stain mitotic spindles during early cleavages, and followed each division with the aid of an autofluorescent compound inside yolk platelets, which differed from the characteristic pink-brownish pigment of the vegetal cytoplasm in zygotes and early embryos. We found that this species exhibits an unequal cleavage characterized by ooplasmic segregation, oblique inclination of mitotic spindles, and differences in size and positioning of the asters in relation to the cellular cortex. Furthermore, we detected asynchrony in cleavage timing between the two large macromeres C and D, which increases the number of cleavage rounds required to reach a particular cell stage in comparison to other spiralians. Here, we report the presence of a transient and previously undescribed U-shaped embryo in this species. The present detailed description of A. califórnica early development deviates considerably from stereotypical patterns described in other spiralians. Our observations demonstrate that early spiralian development can be more plastic than previously thought.

Elysia diomedea , otherwise known as the “Mexican dancer”, aries in adult size and color across its geographical distribution in Ecuador. Because of morphological variation and the absence of genetic information for this species in Ecuador, we analyzed mtDNA sequences in three populations (Ballenita, La Cabuya, and Mompiche) and confirmed that individuals from the three locations belonged to E. diomedea and that there was no population structure that could explain their morphological differences. Next, we analyzed general aspects about the reproductive biology and embryology of this species. Live slugs from the Ballenita population were maintained and reproduced ex situ. Egg ribbons and embryos were fixed and observed by brightfield and confocal microscopy. We observed a single embryo per capsule, 98 embryos per mm2 of egg ribbon, and compared the cleavage pattern of this species to that of other heterobranchs and spiralians. E.diomedea early development was characterized by a slight unequal first cleavage, occurrence of a 3-cell stage in the second cleavage, and the formation of an enlarged second quartet of micromeres. We observed clear yolk bodies in the egg capsules of some eggs ribbons at early stages of develop-ment. Both reproductive and embryological characteristics, such as presence of stomodeum in the larva, and ingestion of particles after hatching confirmed the planktotrophic veliger larvae of this species, consistent with the majority of sacoglossans from the Eastern and Northeast Pacific Oceans..

Considering worldwide amphibian population decline, sperm cryopreservation should be a priority for conservation of species in areas of high biodiversity, such as the Neotropics. In this study, we present the results of two cryopreservation experiments involving Rhinella marina sperm. Freezing was performed in a -80 °C freezer and dimethyl sulfoxide (DMSO) was used as cryoprotective agent. In the first experiment, the effects of 5%, 10%, and 15% DMSO were evaluated in sperm lysis and fertilization capacity. Samples were incubated for 10 minutes at 4 °C before freezing. For thawing, two procedures were tested: 21 °C thawing to be used immediately and 4 °C thawing, to be used two hours later in in vitro fertilizations. The best treatment was 10% DMSO plus thawing at 4 °C, that achieved 20% successful fertilizations. In the second experiment, two solutions were tested: 10% DMSO with and without HEPES. Freezing and post-thawing in vitro fertilizations were performed after a two hour incubation period at 4 °C. A considerable improvement in fertilization percentages was obtained in this experiment, with a 75% for DMSO alone, and a 70% for DMSO + HEPES. These results provide good perspectives for future implementation of sperm cryopreservation in Neotropical institutions for local threatened species.

Current rates of biodiversity loss pose an unprecedented challenge to the conservation community, particularly with amphibians and freshwater fish as the most threatened vertebrates. An increasing number of environmental challenges, including habitat loss, pathogens, and global warming, demand a global response toward the sustainable management of ecosystems and their biodiversity. Conservation Breeding Programs (CBPs) are needed for the sustainable management of amphibian species threatened with extinction. CBPs support species survival while increasing public awareness and political influence. Current CBPs only cater for 10% of the almost 500 amphibian species in need. However, the use of sperm storage to increase efficiency and reliability, along with an increased number of CBPs, offer the potential to significantly reduce species loss. The establishment and refinement of techniques over the last two decades, for the collection and storage of amphibian spermatozoa, gives confidence for their use in CBPs and other biotechnical applications. Cryopreserved spermatozoa has produced breeding pairs of frogs and salamanders and the stage is set for Lifecycle Proof of Concept Programs that use cryopreserved sperm in CBPs along with repopulation, supplementation, and translocation programs. The application of cryopreserved sperm in CBPs, is complimentary to but separate from archival gene banking and general cell and tissue storage. However, where appropriate amphibian sperm banking should be integrated into other global biobanking projects, especially those for fish, and those that include the use of cryopreserved material for genomics and other research.