The QseBC Quorum Sensing System is Involved in Salmonella enterica serovar Typhimurium Colonization of the Swine Gastrointestinal Tract

The response of bacteria to hormone-like, chemical molecules is termed quorum sensing, a mechanism for cell-to-cell communication that includes sensing the host environment. In the gastromtestinal tract, at least two quorum sensing molecules are present that activate the bacterial QseBC quorum sensing system. autoinducer-3 (AI-3) and norepinephrine (NE). Al-3 is produced by bacteria, whereas NE is produced by the host, often during stress. We have demonstrated that the motility of Salmonella enterica serovar Typhimurium is enhanced in the presence of NE and 10% pre-conditioned medium (AI-3) from the wild-type strain

Serological Response of Swine to an Attenuated Salmonella enterica serovar Typhimurium Strain that Reduces Gastrointestinal Colonization, Fecal Shedding and Disease due to Virulent Salmonella Typhimurium

Swine are often asymptomatic carriers of Salmonella spp. Interventions are needed to limit Salmonella colonization of swine to enhance food safety. An attenuated Salmonella enterica serovar Typhimurium mutant strain (BBS 202) was tested in swine to determine whether vaccination could provide protection against wild-type S. Typhimurium challenge. Two groups of piglets (n=14/group) received an intranasal inoculation of BBS 202 or a PBS placebo at 6-weeks of age with a booster 2-weeks later. At 11-weeks of age, all pigs were challenged with the parental, wild-type S. Typhimurium by intranasal inoculation

Differences in Pathogenesis for Salmonella enterica serovar Typhimurium in the Mouse Versus the Swine Model Identifies Bacterial Gene Products Required for Systemic but not Gastrointestinal Disease

Over the last several decades, the mouse model of typhoid fever has been an extremely productive model to investigate Salmonella enterica serovar Typhimurium (S Typhimurium) pathogenesis. The mouse is the paradigm for investigating systemic disease due to infection by Salmonella; however, the swine model of gastrointestinal colonization and enteric disease due to Salmonella is better suited to address food safety. Although certain Salmonella mutants may be attenuated for colonization and disease in both the mouse and swine model, others may only be attenuated in one of the models depending on whether the gene product is required for gastrointestinal or systemic disease. Recent research performed on the swine model in our laboratories with comparison to the literature on the mouse model, illustrates the discrepancy between these two models

Associations of the porcine immune response and genetic polymorphisms with the shedding of Salmonella enterica serovar Typhimurium

A major focus of our collaborattve research is to investigate the porctne response to Infection with Salmonella to 1) identify porctne genes differentially regulated during Infection and 2) Identify and associate genetic polymorphisms within these genes with infection status across swine populations In the current study, 40 crossbred pigs were intranasally inoculated with Salmonella enterica serovar Typhimurium and monitored for Salmonella fecal shedding and blood 1mmune parameters at 2, 7, 14 and 20 days post-inoculation (dpi). Using a multivariate permutation test, a positive correlation was observed between Salmonella shedding and Interferon-gamma (IFNG) levels at 2 and 7 dpi (p\u3c0.05), with a greater number of Salmonella shedding 1n the animals with higher IFNG levels

Correlating blood immune parameters and a CCT7 genetic variant with the shedding of Salmonella enterica serovar Typhimurium in swine

The porcine response to Salmonella infection is critical for control of Salmonella fecal shedding and the establishment of Salmonella carrier status. In this study, 40 crossbred pigs were intranasally inoculated with Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) and monitored for Salmonella fecal shedding and blood immune parameters at 2, 7, 14 and 20 days post-inoculation (dpi). Using a multivariate permutation test, a positive correlation was observed between Salmonella Typhimurium shedding levels at 2 and 7 dpi and serum interferon-gamma (IFNγ) levels at 2 dpi (p \u3c 0.05), with Salmonella being shed in greater numbers from animals with higher IFNγ levels. A positive correlation was also observed between IFNγ levels and the number of banded neutrophils (2 dpi), circulating neutrophils (7 and 14 dpi), monocytes (7 dpi), and white blood cells (WBCs) (7, 14 and 20 dpi). We have further performed association studies on these immune response parameters as well as shedding status of the Salmonella-infected pigs with a single nucleotide polymorphism (SNP) in the porcine gene CCT7, previously shown by our group to be transcriptionally up-regulated in swine experimentally inoculated with Salmonella Typhimurium. Our analyses with the 40 pigs suggest a positive association (p = 0.0012) of SNP genotype A/G at position AK240296.c1153G \u3e A of the CCT7 gene with Salmonella shedding at 7 dpi compared to the G/G homozygote genotype. Linking specific genes and genetic polymorphisms with the porcine immune response to Salmonella infection and shedding may identify potential markers for carrier pigs as well as targets for disease diagnosis, intervention and prevention

Direct Bacterial Killing In Vitro by Recombinant Nod2 Is Compromised by Crohn's Disease-Associated Mutations

Background: A homeostatic relationship with the intestinal microflora is increasingly appreciated as essential for human health and wellbeing. Mutations in the leucine-rich repeat (LRR) domain of Nod2, a bacterial recognition protein, are associated with development of the inflammatory bowel disorder, Crohn’s disease. We investigated the molecular mechanisms underlying disruption of intestinal symbiosis in patients carrying Nod2 mutations. Methodology/Principal Findings: In this study, using purified recombinant LRR domains, we demonstrate that Nod2 is a direct antimicrobial agent and this activity is generally deficient in proteins carrying Crohn’s-associated mutations. Wildtype, but not Crohn’s-associated, Nod2 LRR domains directly interacted with bacteria in vitro, altered their metabolism and disrupted the integrity of the plasma membrane. Antibiotic activity was also expressed by the LRR domains of Nod1 and other pattern recognition receptors suggesting that the LRR domain is a conserved anti-microbial motif supporting innate cellular immunity. Conclusions/Significance: The lack of anti-bacterial activity demonstrated with Crohn’s-associated Nod2 mutations in vitro, supports the hypothesis that a deficiency in direct bacterial killing contributes to the association of Nod2 polymorphism

Sensing and Adaptation to Low pH Mediated by Inducible Amino Acid Decarboxylases in Salmonella

During the course of infection, Salmonella enterica serovar Typhimurium must successively survive the harsh acid stress of the stomach and multiply into a mild acidic compartment within macrophages. Inducible amino acid decarboxylases are known to promote adaptation to acidic environments. Three low pH inducible amino acid decarboxylases were annotated in the genome of S. Typhimurium, AdiA, CadA and SpeF, which are specific for arginine, lysine and ornithine, respectively. In this study, we characterized and compared the contributions of those enzymes in response to acidic challenges. Individual mutants as well as a strain deleted for the three genes were tested for their ability (i) to survive an extreme acid shock, (ii) to grow at mild acidic pH and (iii) to infect the mouse animal model. We showed that the lysine decarboxylase CadA had the broadest range of activity since it both had the capacity to promote survival at pH 2.3 and growth at pH 4.5. The arginine decarboxylase AdiA was the most performant in protecting S. Typhimurium from a shock at pH 2.3 and the ornithine decarboxylase SpeF conferred the best growth advantage under anaerobiosis conditions at pH 4.5. We developed a GFP-based gene reporter to monitor the pH of the environment as perceived by S. Typhimurium. Results showed that activities of the lysine and ornithine decarboxylases at mild acidic pH did modify the local surrounding of S. Typhimurium both in culture medium and in macrophages. Finally, we tested the contribution of decarboxylases to virulence and found that these enzymes were dispensable for S. Typhimurium virulence during systemic infection. In the light of this result, we examined the genomes of Salmonella spp. normally responsible of systemic infection and observed that the genes encoding these enzymes were not well conserved, supporting the idea that these enzymes may be not required during systemic infection

Differential Gene Expression by RamA in Ciprofloxacin-Resistant Salmonella Typhimurium

Overexpression of ramA has been implicated in resistance to multiple drugs in several enterobacterial pathogens. In the present study, Salmonella Typhimurium strain LTL with constitutive expression of ramA was compared to its ramA-deletion mutant by employing both DNA microarrays and phenotype microarrays (PM). The mutant strain with the disruption of ramA showed differential expression of at least 33 genes involved in 11 functional groups. The study confirmed at the transcriptional level that the constitutive expression of ramA was directly associated with increased expression of multidrug efflux pump AcrAB-TolC and decreased expression of porin protein OmpF, thereby conferring multiple drug resistance phenotype. Compared to the parent strain constitutively expressing ramA, the ramA mutant had increased susceptibility to over 70 antimicrobials and toxic compounds. The PM analysis also uncovered that the ramA mutant was better in utilization of 10 carbon sources and 5 phosphorus sources. This study suggested that the constitutive expression of ramA locus regulate not only multidrug efflux pump and accessory genes but also genes involved in carbon metabolic pathways

Survival of Escherichia coli in the environment: fundamental and public health aspects

In this review, our current understanding of the species Escherichia coli and its persistence in the open environment is examined. E. coli consists of six different subgroups, which are separable by genomic analyses. Strains within each subgroup occupy various ecological niches, and can be broadly characterized by either commensalistic or different pathogenic behaviour. In relevant cases, genomic islands can be pinpointed that underpin the behaviour. Thus, genomic islands of, on the one hand, broad environmental significance, and, on the other hand, virulence, are highlighted in the context of E. coli survival in its niches. A focus is further placed on experimental studies on the survival of the different types of E. coli in soil, manure and water. Overall, the data suggest that E. coli can persist, for varying periods of time, in such terrestrial and aquatic habitats. In particular, the considerable persistence of the pathogenic E. coli O157:H7 is of importance, as its acid tolerance may be expected to confer a fitness asset in the more acidic environments. In this context, the extent to which E. coli interacts with its human/animal host and the organism's survivability in natural environments are compared. In addition, the effect of the diversity and community structure of the indigenous microbiota on the fate of invading E. coli populations in the open environment is discussed. Such a relationship is of importance to our knowledge of both public and environmental health. The ISME Journal (2011) 5, 173-183; doi:10.1038/ismej.2010.80; published online 24 June 2010NATO [ESP.EAP.CLG 981785]; The Soil Biotechnology Foundationinfo:eu-repo/semantics/publishedVersio