No abstract

Amphibians support symbiotic bacterial communities on their skin that protect against a range of infectious pathogens, including the amphibian chytrid fungus. The conditions under which amphibians are maintained in captivity (e.g. diet, substrate, enrichment) in ex situ conservation programmes may affect the composition of the bacterial community. In addition, ex situ amphibian populations may support different bacterial communities in comparison to in situ populations of the same species. This could have implications for the suitability of populations intended for reintroduction, as well as the success of probiotic bacterial inoculations intended to provide amphibians with a bacterial community that resists invasion by the chytrid fungus. We aimed to investigate the effect of a carotenoid-enriched diet on the culturable bacterial community associated with captive red-eyed tree frogs (Agalychnis callidryas) and make comparisons to bacteria isolated from a wild population from the Chiquibul Rainforest in Belize. We successfully showed carotenoid availability influences the overall community composition, species richness and abundance of the bacterial community associated with the skin of captive frogs, with A. callidryas fed a carotenoid-enriched diet supporting a greater species richness and abundance of bacteria than those fed a carotenoid-free diet. Our results suggest that availability of carotenoids in the diet of captive frogs is likely to be beneficial for the bacterial community associated with the skin. We also found wild A. callidryas hosted more than double the number of different bacterial species than captive frogs with very little commonality between species. This suggests frogs in captivity may support a reduced and diverged bacterial community in comparison to wild populations of the same species, which could have particular relevance for ex situ conservation projects.

Community genetics examines how genotypic variation within a species influences the associated ecological community. The inclusion of additional environmental and genotypic factors is a natural extension of the current community genetics framework. However, the extent to which the presence of and genetic variation in associated species influences interspecific interactions (i.e., genotype x genotype x environment [G x G x E] interactions) has been largely ignored. We used a community genetics approach to study the interaction of barley and aphids in the absence and presence of rhizosphere bacteria. We designed a matrix of aphid genotype and barley genotype combinations and found a significant G x G x E interaction, indicating that the barley-aphid interaction is dependent on the genotypes of the interacting species as well as the biotic environment. We discuss the consequences of the strong G x G x E interaction found in our study in relation to its impact on the study of species interactions in a community context.

Transfectants with a wide range of cellular phenotypes are obtained during the process of cell line generation. For the successful manufacture of a therapeutic protein, a means is required to identify a cell line with desirable growth and productivity characteristics from this phenotypically wide-ranging transfectant population. This identification process is on the critical path for first-in-human studies. We have stringently examined a typical selection strategy used to isolate cell lines suitable for cGMP manufacturing. One-hundred and seventy-five transfectants were evaluated as they progressed through the different assessment stages of the selection strategy. High producing cell lines, suitable for cGMP manufacturing, were identified. However, our analyses showed that the frequency of isolation of the highest producing cell lines was low and that ranking positions were not consistent between each assessment stage, suggesting that there is potential to improve upon the strategy. Attempts to increase the frequency of isolation of the 10 highest producing cell lines, by in silico analysis of alternative selection strategies, were unsuccessful. We identified alternative strategies with similar predictive capabilities to the typical selection strategy. One alternate strategy required fewer cell lines to be progressed at the assessment stages but the stochastic nature of the models means that cell line numbers are likely to change between programs. In summary, our studies illuminate the potential for improvement to this and future selection strategies, based around use of assessments that are more informative or that reduce variance, paving the way to improved efficiency of generation of manufacturing cell lines.

Ex situ conservation is an important tool for the prevention of species extinction in amphibians. Currently, there is limited information on the nutritional requirements of amphibians in captivity, and there have been anecdotal reports of skin colour degradation in captive amphibians. Amphibians use carotenoids for skin pigmentation, and because carotenoids are only obtainable through the diet, colour degradation could result from limited carotenoid availability. Studies of other vertebrate taxa have shown that carotenoids contribute to health and reproductive success; however, their importance to amphibians is currently unknown. We assessed the influence of carotenoids on the health and development of red‐eye tree frogs (Agalychnis callidryas) during larval and post‐metamorphic stages, and looked at effects on skin colour and reproductive success. Carry‐over effects of larval exposure to carotenoids were also investigated, and the effects of carotenoids on skin colour development and degradation in adulthood were examined. Carotenoids did not significantly influence larval growth or survival; however, post‐metamorphic carotenoid availability significantly increased growth rate in female but not in male frogs. Frogs fed a carotenoid diet post‐metamorphosis had significantly redder skin than controls, and larval carotenoid exposure significantly influenced post‐metamorphic skin colour. Fecundity was significantly higher in female frogs raised on a carotenoid diet post‐metamorphosis compared with controls. Finally, skin colour did not change in adulthood despite changes in dietary carotenoid availability, which suggests that there is a critical period during post‐metamorphic growth for deposition of carotenoids in the skin. We have shown that carotenoids influence the development, phenotype and reproductive success of A. callidryas, and these important nutrients should therefore be considered when nutritional recommendations for amphibians are made.