Streptomycin-resistant virulent Salmonella enterica serovar Typhimurium SL1344 was used for all infection experiments. Properties of disease-specific and commonly expressed genes.Twenty-seven of 50 Salmonella promoters (56%) with very high levels of activity during infection drive expression of genes with no homologue in the closely related species E. coli, which was more than expected (32% of all annotated Salmonella genes lack E. coli homologues). arizonae are especially rare, but it may affect immunocompromised patients and infants. Identification of Salmonella promoters with high levels of activity during enteritis.To identify Salmonella promoters with high levels of activity in the gut, a diverse plasmid-based Salmonella GFP-promoter trap library was FACS sorted for high fluorescence in the cecal contents of streptomycin-pretreated, intragastrically infected mice. ], Weldin, J.C. "The colon-typhoid group of bacteria and related forms. Subspecies 1 of Salmonella enterica is responsible for almost all Salmonella infections of warm-blooded animals. We present a case of endocarditis in a patient with sickle cell disease and a review of earlier cases caused by this rare human pathogen. Numerous studies have investigated pathogenic in vivo gene expression by diverse techniques such as in vivo expression technology (IVET), differential fluorescence induction, in vivo-induced antigen technology, and microarray transcriptome analysis (9, 38, 54, 56, 58, 60, 62, 66, 72, 74), but these studies have mostly focused on genes that are specifically induced during infection, whereas genes with high levels of in vitro activity have been mostly disregarded. Ninety-six recovered clones were characterized by PCR restriction fragment length polymorphism, revealing 34 nonredundant clones. PLOS Neglected Tropical Diseases, Vol. While nutrient utilization and energy conversion appeared to dominate Salmonella transcription during enteritis, virulence factor transcription seemed to dominate during typhoid fever. In this study, we compared the genomes of outbreak associated non-outbreak causing Salmonella ser. For example, Salmonella enterica subspecies enterica serotype Enteritidis can be referred to simply Salmonella Enteritidis, nomenclature that will be used throughout this chapter. Scientists classify Salmonella into serotypes (types) by identifying structures on the bacteria’s surfaces. HilD is a positive regulator in a regulatory cascade that leads to expression of SPI-1-associated genes, which are required for induction of enteritis but not during systemic infection. ... Salmonella enterica subsp. Int. 1B; data not shown) and emission ratios typical for GFP (14), indicating that GFP synthesis and at least partial fluorophore formation occurred in the gut. In particular, we generated a mutant lacking the global anaerobiosis regulator fnr to test the combined relevance of several identified Fnr-dependent genes associated with anaerobic respiration and mixed acid fermentation that were selectively expressed during enteritis. However, the functional relevance of these negative flagella gene regulators in the murine colitis model remained unclear, since we also found high levels of enteric expression of the flhDC operon encoding the master regulator of class II flagellar genes and expression of the major flagellin gene fliC (data not shown). J. Syst. In addition, an IVET study has revealed intestinal fhuA expression (38). 2A). Iowa State J. Sci. Reference: How to cite this resource - Schoch CL, et al. Indeed, the highly expressed genes play an outstanding role for Salmonella virulence, since at least 16 (32%) of the identified 50 operons with very high levels of expression during infection encoded genes with essential functions in one or both disease models (Tables 1, 2, and 4). enterica serovar Typhimurium var. A chorismate mutase isozyme, which is a part of pheA, was highly expressed during typhoid fever (Table 2) and might compensate for deficient aroQ. Salmonella enterica serovar Typhi can cause life-threatening bacterial infections called typhoid fever. Int. The yaoF mutant was attenuated in the typhoid fever model, where it was specifically expressed, but not during enteritis. There are only few cases of salmonella … A new form of Salmonella Typhimurium (ST313) emerged in the southeast of the African continent 75 years ago, followed by a second wave which came out of central Africa 18 years later. as the neotype species of the genus Salmonella Lignieres 1900 In contrast to this enteritis model, mice that had not received a streptomycin pretreatement had no fluorescent Salmonella in their ceca 1 day after intragastric infection with initially nonfluorescent Salmonella carrying the PrpsA-gfp_ova fusion, suggesting an absence of de novo GFP biosynthesis. Regulations in most jurisdictions in the United States exclude individuals who … Int. The prevailing dogma in murine S. enterica serovar Typhimurium pathogenesis is that distinct virulence mechanisms— Salmonella … (B) Coefficient of variation of results obtained from repeated measurements of individual promoter-gfp_ova fusions in typhoid fever (spleen) and enteritis (cecum) models. Determination of GFP and GFP_OVA fluorophore formation and degradation. rev. [No PubMed record available. The bacteria are located in a special membrane compartment, the so-called Salmonella-containing vacuole (SCV). n.h., homology below 5%. Salmonella enterica serovar Heidelbergis one of the common invasive nontyphoidal Salmonellaassociated with greater risk for severe disease [4, 5]. Background: Most publicly available genomes of Salmonella enterica are from human disease in the US and the UK, or from domesticated animals in the US. enterica serotype Dublin is a host-adapted serotype predominantly found in cattle and occasionally in swine, sheep, horses, and zoological animals (5, 7, 11, 50).Salmonellosis in animals always presents a potential zoonotic threat. Three transcriptional fusions were located within regions of the Salmonella genome sequence that lack annotated genes in the right orientation (fragment I, nucleotides 1656707 to 1656128 [nucleotides in the genome sequence]; fragment II, nucleotides 2807962 to 2807445; fragment III, nucleotides 1334351 to 1333800), and these were excluded from further analysis (see Results). ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology. 3). nov., nom. The residual four promoters might be active but not strong enough to overcome the high detection threshold for single-copy reporter constructs. To determine virulence defects, groups of five streptomycin-pretreated mice were infected intragastrically (enteritis) or groups of five untreated mice were infected systemically (typhoid fever) with a mixture of wild-type and mutant Salmonella strains. Salmonella enterica subsp. Surprisingly, two virulence-associated promoters were found to be active in both disease models, and one of these (PhilD) was independently identified in nonredundant clones in both disease models. We also tested mutants lacking the putative hemolysin yaoF or the periplasmic chorismate mutase aroQ, which were both selectively expressed during typhoid fever, and a mutant that lacked the riboflavin biosynthesis gene ribB with high expression in both disease models. The competitive index (CI) was calculated as (output mutant/wild-type ratio)/(input mutant/wild-type ratio). [No PubMed record available. Philos. Salmonella enterica serovar Pullorum is the causative agent of pullorum disease in poultry, an acute systemic disease more common in young birds. American Society for Microbiology, Washington, D.C. (1992). Journal of Microbiology & Biology Education, Microbiology and Molecular Biology Reviews. Utilization of unstable GFP variants such as GFP_OVA decreases sensitivity and thus allows selective identification of very strong promoters (61, 76). The high redundancy of the sorted sublibrary (see above) might suggest substantial coverage of such promoters. We aimed to date the emergence of this resistance in historical isolates of S enterica serotype Typhimurium. After 100-fold dilution into PBS, GFP_OVA-expressing Salmonella cells were sorted by two-color flow cytometry using a biosafety level 2-equipped high-speed sorter (FacsDIVA; BD Biosciences) as previously described (14, 61). Opinion 80 ." The cumulative activity of promoters associated with nutrient utilization and energy conversion (UTIL), SPI-1, SPI-2, or the PhoPQ regulon (PhoP) is also shown. As a consequence of low GC content, these genes had a markedly different codon usage compared to “classical” highly expressed genes such as ribosomal proteins and chaperones, resulting in poor codon adaptation indices (reference 25 and data not shown). In some patients the infection spreads … Bacterial strains, Salmonella promoter trap library, and construction of mutants. Surprisingly, the data reveal that instead of protein biosynthesis genes, disease-specific genes such as Salmonella pathogenicity island 1 (SPI-1)-associated genes and genes involved in anaerobic respiration (enteritis) or SPI-2-associated genes and genes of the PhoP regulon (typhoid fever), respectively, dominate Salmonella in vivo gene expression. Salmonella strains sometimes cause infection in urine, blood, bones, joints, or the nervous system (spinal fluid and brain), and can cause severe disease. Extracted Salmonella DNA was amplified/detected using real-time PCR on an ABI 7500 instrument (Thermo Fisher, Waltham, MA) with a kit of analyte-specific reagents for detection of S. enterica (Liferiver). 2A. [] To date, more than 2500 serovars of S enterica have been described. source for nomenclature or classification - please consult the The riboflavin biosynthesis gene ribB was essential in both disease models, in agreement with its expression pattern. [No PubMed record available.]. As a critical precondition for this approach, we demonstrated that GFP fluorescence can be used to monitor Salmonella gene expression in the gut despite low oxygen availability. For six of these constructs, GFP expression in the native genomic context and copy number was sufficiently high for detection in duplicate experiments (Table 3), confirming the successful identification of promoters with exceptionally high in vivo activity (35). (1987) 37:465-468. (1984) (reference relating to orthographic revision), Skerman VBD et al. These data represented only a first coarse estimate of the functional profile of highly in vivo-expressed Salmonella genes because of the limited number of promoter fusions and replicate measurements, as well as potentially distorting effects of copy number, topology, and position of the fusion junctions. J. Syst. ], Lapage, S.P., Sneath, P.H.A., Lessel, E.F., Skerman, V.B.D., Seeliger, H.P.R., and Clark, W.A. nov., nom. A formate transporter (FocA) and the pyruvate formiate lyase (PflB) are involved in mixed acid fermentation during anaerobic growth of Salmonella, which is consistent with oxygen limitation in the gut. ], Translation table 11 (Bacterial, Archaeal and Plant Plastid), Hill LR et al. The plots shows ranges (whiskers) and quartiles (the boxes extend from the 25th percentile to the 75th percentile, and the lines within boxes depict the medians). enterica Salmonella enterica subsp. Infection begins with the ingestion of contaminated food or water so that salmonellae reach the intestinal epithelium and trigger gastrointestinal disease. Extensive in vivo gene expression data have been obtained for various pathogens; these data could be used for testing the relevance of protein biosynthesis genes during infection, but most studies focused on genes specifically induced during infection, while constitutively expressed genes were mostly disregarded. Salmonella enterica is an important enteric pathogen of humans and a variety of domestic and wild animals. 2A) lack E. coli homologues, whereas most commonly expressed genes (black circles) are conserved in E. coli. pheA encodes the bifunctional chorismate mutase/prephenate dehydratase involved in phenylalanine biosynthesis. Salmonella enterica serovar Typhi is an invasive … Evol. The results indicate an unexpected dominance of disease-specific genes and a marked shift in the overall functional profiles during enteritis and systemic disease. Outbreaks can occur, and there was a recent multistate foodborne outbreak in Australia. (formerly Salmonella choleraesuis) SYNONYM OR CROSS REFERENCE: ... malignancy, corrhosis, chronic granulomatous disease, sickle cell disease, lymphoproliferative disease, or collagen vascular disease) have a higher risk of developing bacteremia due to a Salmonella infection (4, 7). Many of these genes share a strongly biased codon usage, and this has been exploited to predict highly expressed genes in both environmental and pathogenic bacteria (44, 46). Invasive Non-typhoidal Salmonella Infections in Asia: Clinical Observations, Disease Outcome and Dominant Serovars from an Infectious Disease Hospital in Vietnam. Salmonella thus appeared to adapt to different disease conditions, mainly by using genes that lack properties of “typical” highly expressed genes. To compare Salmonella gene expression during enteritis and typhoid fever, we also included some of our previously identified typhoid fever-associated promoters (61). In the present study, we used only those fusions that were obtained from a single sort cycle for high activity in vivo, while the other original sort cycles selecting for certain in vitro activities in LB culture were disregarded. The SPI-1-associated rtsA gene is cotranscribed with rtsB, which encodes a negative regulator of flagellum synthesis (26). Database (Oxford). Enterica Salmonella Choleraesuis causes a necrotizing fibrinous pneumonia similar to porcine pleuropneumonia, and Salmonella Typhisuis causes a chronic suppurative bronchopneumonia. Salmonella Enteritidis is a rod-shaped, gram-negative, proteobacteria that is non-motile [Figure 4]. Salmonella enterica subspecies enterica has 2610 different serotypes; the most well known being serotypes Typhi, Paratyphi, Enteriditis, Typhimurium and Choleraesuis (1). While protein biosynthesis is essential for bacterial growth under all conditions, these data suggested that it might not be the dominating activity in Salmonella during infection. SlyA itself is another transcriptional regulator (13). In subacute outbreaks, there are dead-in-shell embryos, or dead chicks on the … Fowl typhoid (Salmonella gallinarum infection): S. gallinarum causes disease mainly in adult or growing chickens and turkeys but also affects ducks, pheasants, guinea fowl, peafowl, grouse, and quail.Acute cases of infection are related to septicemia. The enteric expression was verified with a GFP insertion in the native chromosomal locus (Table 3). Salmonella infection (salmonellosis) is a common bacterial disease that affects the intestinal tract. Salmonella enterica serotype Heidelberg is primarily a poultry adapted serotype of Salmonella that can also colonize other hosts and cause human disease. (A) Mice were infected with initially nonfluorescing Salmonella cells from a stationary in vitro culture. With these limitations in mind, the overall analysis revealed a striking functional difference between the two different disease models. Int. We used an episomal GFP_OVA promoter trap library in SL1344 with about 20-fold genome coverage that was previously described (61). As a result, for some promoters we had to measure fusions other than those of the previous study. One day later, cecal contents were prepared as described above and pooled. rev. Salmonella enterica subsp. 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