Orally administered phages to control zoonotic pathogens face important challenges, mainly related to the hostile conditions found in the gastrointestinal tract (GIT). These include temperature, salinity and primarily pH, which is exceptionally low in certain compartments. Phage survival under these conditions can be jeopardized and undermine treatment. Strategies like encapsulation have been attempted with relative success, but are typically complex and require several optimization steps. Here we report a simple and efficient alternative, consisting in the genetic engineering of phages to display lipids on their surfaces. Escherichia coli phage T7 was used as a model and the E. coli PhoE signal peptide was genetically fused to its major capsid protein (10 A), enabling phospholipid attachment to the phage capsid. The presence of phospholipids on the mutant phages was confirmed by High Performance Thin Layer Chromatography, Dynamic Light Scattering and phospholipase assays. The stability of phages was analysed in simulated GIT conditions, demonstrating improved stability of the mutant phages with survival rates 102–107 pfu.mL−1 higher than wild-type phages. Our work demonstrates that phage engineering can be a good strategy to improve phage tolerance to GIT conditions, having promising application for oral administration in veterinary medicine.
Guimarães is a UNESCO world heritage site, deemed as such in 2001 and hosted the 2012 European Capital of Culture. This paper examines the profile, destination image, and motivations of tourists’ visiting behaviors in Guimarães before the cultural event. Based on survey responses from 276 tourists, we found that tourists arriving in Guimarães had previously visited the two most important cities in the northern part of Portugal. These respondents were relatively young and well educated compared to the average tourists that visited Portugal. The results suggest that many tourists were aware of the city status as a world heritage site that encompasses a historic center, monuments, and architectural buildings. Furthermore, these perceptions shape the image of Guimarães, as the factor analysis indicates that “historical background and functionality” was the most reliable and valid factor behind the choice of visiting the city.
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