Project description:Salmonella enterica variants exhibit diverse host adaptation, outcome of infection, and associated risk to humans. Analysis of 6,335 Salmonella isolates recovered from integrated human-animal surveillance in Emilia Romagna region, Northern Italy, (human population ca 4,500,000), from 2012 to 2017 showed that Salmonella enterica serovar Derby constitutes a swine associated serovar in this epidemiological context while representing also a significant causative agent of human infections. Comparison of the distribution of subtypes of Salmonella Derby from human and swine identified isolates with a distinct PFGE profile that were significantly less isolated in human infections than in swine infections compared to all other subtypes. Here we show that isolates with this PFGE profile form a distinct phylogenetic sub-clade within Salmonella Derby and exhibit a marked reduction in invasion and replication in human epithelial cells but a relatively small reduction in swine epithelial cells, in line with the epidemiological evidence. A single missense mutation in hilD, that encodes the master-regulator of the Salmonella Pathogenicity Island 1 (SPI-1), was identified in this lineage of Salmonella Derby. Since SPI-1 encodes for a primary system of Salmonella invasion into epithelial cells, we investigated the role of the observed mutation in detail. We demonstrated that the missense mutation results in a loss of function of HilD that accounts for the reduced invasion and replication in human epithelial cells while showing a relatively small impact on the interaction with swine cells. This finding is suggestive of a mechanism of invasion alternative to SPI-1 in the Salmonella-swine combination
Project description:Bifidobacterium thermophilum RBL67 (RBL67), a human fecal isolate and promising probiotic candidate, showed antagonistic and protective effects against Salmonella and Listeria in vitro. However, the underlying mechanisms fostering these health-related effects remain unknown. Therefor the transcriptome response of RBL67 and Salmonella enterica subsp. enterica serovar Typhimurium N-15 (N-15) in co-culture compared to the response in their respective mono-cultures. RNA was extracted from culture samples taken after 4 (N-15) or 5 h (RBL67) and RNAseq was performed on an Illumina HiSeq 2000 sequencer. Three biological replciates were performed resulting in 12 data sets: 3 RBL67 mono culture, 3 N15 mono-culture, 3 RBL67 co-culture, 3 N15 co-culture. Our study provided first insights into probiotic-pathogen interaction on transcriptional level and suggests a mechanism for how probiotic organisms can protect the host from infections.
Project description:Infection with Salmonella enterica serovar Typhi in humans causes the systemic, life-threatening disease typhoid fever. In the laboratory, typhoid fever can be modeled through the inoculation of susceptible mice with Salmonella enterica serovar Typhimurium. The ensuing disease is characterized by systemic dissemination and colonization of many organs, including the liver, spleen and gallbladder. Using this murine model, we previously characterized the interactions between Salmonella Typhimurium and host cells in the gallbladder and showed that this pathogen can successfully invade gallbladder epithelial cells and proliferate. Additionally, we showed that Salmonella Typhimurium can use bile phospholipids to grow at high rates. These abilities are likely important for quick colonization of the gallbladder during typhoid fever and further pathogen dissemination through fecal shedding. To further characterize the interactions between Salmonella and the gallbladder environment we compared the transcriptome of Salmonella cultures grown in LB or physiological murine bile. Our data showed that many genes involved in bacterial central metabolism are affected by bile, with the citric acid cycle being repressed and alternative respiratory systems being activated. Additionally, our study revealed a new aspect of Salmonella interactions with bile through the identification of phoP as a bile-responsive gene. Repression of phoP expression does not involve PhoPQ sensing of a bile component. Due to its critical role in Salmonella virulence, further studies in this area will likely reveal aspects of the interaction between Salmonella and bile that are relevant to disease.