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Abstract BackgroundFuture expansion of corn-derived ethanol raises concerns of sustainability and competition with the food industry. Therefore, cellulosic biofuels derived from agricultural waste and dedicated energy crops are necessary. To date, slow and incomplete saccharification as well as high enzyme costs have hindered the economic viability of cellulosic biofuels, and while approaches like simultaneous saccharification and fermentation (SSF) and the use of thermotolerant microorganisms can enhance production, further improvements are needed. Cellulosic emulsions have been shown to enhance saccharification by increasing enzyme contact with cellulose fibers. In this study, we use these emulsions to develop an emulsified SSF (eSSF) process for rapid and efficient cellulosic biofuel production and make a direct three-way comparison of ethanol production betweenS. cerevisiae,O. polymorpha, andK. marxianusin glucose and cellulosic media at different temperatures. ResultsIn this work, we show that cellulosic emulsions hydrolyze rapidly at temperatures tolerable to yeast, reaching up to 40-fold higher conversion in the first hour compared to microcrystalline cellulose (MCC). To evaluate suitable conditions for the eSSF process, we explored the upper temperature limits for the thermotolerant yeastsKluyveromyces marxianusandOgataea polymorpha, as well asSaccharomyces cerevisiae, and observed robust fermentation at up to 46, 50, and 42 °C for each yeast, respectively. We show that the eSSF process reaches high ethanol titers in short processing times, and produces close to theoretical yields at temperatures as low as 30 °C. Finally, we demonstrate the transferability of the eSSF technology to other products by producing the advanced biofuel isobutanol in a light-controlled eSSF using optogenetic regulators, resulting in up to fourfold higher titers relative to MCC SSF. ConclusionsThe eSSF process addresses the main challenges of cellulosic biofuel production by increasing saccharification rate at temperatures tolerable to yeast. The rapid hydrolysis of these emulsions at low temperatures permits fermentation using non-thermotolerant yeasts, short processing times, low enzyme loads, and makes it possible to extend the process to chemicals other than ethanol, such as isobutanol. This transferability establishes the eSSF process as a platform for the sustainable production of biofuels and chemicals as a whole. 

Worldwide, amphibian populations have been declining rapidly. This decline can be attributed to many factors including climate change, pesticide exposure, and emerging infectious diseases, among other important factors, but few studies have examined the influence of species interactions. In this study, we examined how habitat factors and co-occurring avian and mammalian species, as well as humans, exert direct and indirect effects on Neotropical amphibian population dynamics. We further examined how these habitat and species interactions could affect our ability to reliably detect amphibian presence to robustly estimate population trends. We conducted amphibian visual encounter surveys at 26 randomly selected sites in the La Selva Biological Station, in northeastern Costa Rica, as well as 26 sites across five additional forest fragments in the region. Furthermore, we used camera traps to collect data on avian and mammalian communities and human visitation at those amphibian survey plots. From these data, we were able to estimate species occupancy probabilities for leaf litter frogs across sites and their relationships to habitat and interspecific species interaction covariates. We also conducted an experiment with plastic model frogs to estimate detection probabilities when a population is known to occur at a site with certainty. Our results suggested that strawberry poison dart frog ( Oophagapumilio ) occupancy was positively related to secondary forest and their detection was negatively related to increasing air temperatures at the times of the surveys. Leaf litter frog occupancy was negatively related to core La Selva sites and human detections at sites, yet their detection was positively related to human trail presence, which might be related to reduced leaf litter cover due to heavy trampling. Our experimental surveys suggested that Neotropical leaf litter frog communities are difficult to detect when present and future studies should explicitly account for this detection bias to effectively monitor population trends.