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Co-cultivation in microfluidic droplets has emerged as a versatile tool for the study of natural and synthetic microbial communities. In particular, the identification and characterization of syntrophic interactions in these communities is attracting increasing interest due to their critical importance for the functioning of environmental and host-associated communities as well as new biotechnological applications. However, one critical parameter in droplet-enabled co-cultivation that has evaded appropriate evaluation is the droplet size. Given the same number of initial cells, a larger droplet size can increase the length scale secreted metabolites must diffuse as well as dilute the initial concentration of cells and exchanged metabolites, impacting the community dynamics. To evaluate the effect of droplet size on a spectrum of syntrophic interactions, we cultivated a synthetic model system consisting of twoE.coliauxotrophs, whose interactions could be modulated through supplementation of related amino acids in the medium. Our results demonstrate that the droplet size impacts substantially numerous aspects of the growth of a cross-feeding bi-culture, particularly the growth capacity, maximum specific growth rate, and lag time, depending on the degree of the interaction. This work heavily suggests that one droplet size does not fit all types of interactions; this parameter should be carefully evaluated and chosen in experimental studies that aim to utilize droplet-enabled co-cultivation to characterize or elucidate microbial interactions. 

Abstract A first synthesis of available data for the period of Rome’s expansion in Italy (about 400–29 b.c.e.) shows the role of climate and environment in early Roman imperialism. Although global indices suggest a warmer phase with relatively few short-term climate events occuring around the same time as the expansion, local data emphasize the highly variable timing and expression of these trends. This variability casts doubt on ideas of a unitary, historically consequential “Roman Warm Period.” The historical importance of climate and environment to socioeconomic development merits emphasis, but should be understood in terms of evolving, contingent forms of resilience and risk-mitigating behavior by Italian communities during Roman expansion.