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At least 253 records · Page 14Linked to original sources
PubMed1992

Identification and molecular characterization of a Salmonella typhimurium gene involved in triggering the internalization of salmonellae into cultured epithelial cells.

Penetration of intestinal epithelial cells is an important step in the pathogenesis of Salmonella infections. We have characterized a gene, invE, that is necessary for Salmonella invasion of cultured epithelial cells. The predicted amino acid sequence of InvE showed significant homology to the Yersinia outer membrane protein YopN (LcrE). Strains of Salmonella carrying mutations in invE were unable to penetrate Henle-407 human intestinal cells and Madin-Darby canine kidney cells, although they were fully capable of attaching to the same cells. Unlike wild-type Salmonella typhimurium, invE mutants failed to change the intracellular free calcium levels or the distribution of polymerized actin in cultured epithelial cells; neither did they alter the normal architecture of the microvilli of polarized Madin-Darby canine kidney cells. Wild-type S. typhimurium was able to rescue the invasive phenotype of the invE mutants in simultaneous infections of cultured epithelial cells although it did not rescue the Escherichia coli strain RDEC-1. We hypothesize that invE mutants are deficient in triggering the intracellular events that lead to bacterial internalization.

Actin Cytoskeleton↗
PubMed2006

Autophagy controls Salmonella infection in response to damage to the Salmonella-containing vacuole.

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a facultative intracellular pathogen that causes disease in a variety of hosts. S. Typhimurium actively invade host cells and typically reside within a membrane-bound compartment called the Salmonella-containing vacuole (SCV). The bacteria modify the fate of the SCV using two independent type III secretion systems (TTSS). TTSS are known to damage eukaryotic cell membranes and S. Typhimurium has been suggested to damage the SCV using its Salmonella pathogenicity island (SPI)-1 encoded TTSS. Here we show that this damage gives rise to an intracellular bacterial population targeted by the autophagy system during in vitro infection. Approximately 20% of intracellular S. Typhimurium colocalized with the autophagy marker GFP-LC3 at 1 h postinfection. Autophagy of S. Typhimurium was dependent upon the SPI-1 TTSS and bacterial protein synthesis. Bacteria targeted by the autophagy system were often associated with ubiquitinated proteins, indicating their exposure to the cytosol. Surprisingly, these bacteria also colocalized with SCV markers. Autophagy-deficient (atg5-/-) cells were more permissive for intracellular growth by S. Typhimurium than normal cells, allowing increased bacterial growth in the cytosol. We propose a model in which the host autophagy system targets bacteria in SCVs damaged by the SPI-1 TTSS. This serves to retain intracellular S. Typhimurium within vacuoles early after infection to protect the cytosol from bacterial colonization. Our findings support a role for autophagy in innate immunity and demonstrate that Salmonella infection is a powerful model to study the autophagy process.

Animals↗
PubMed2006

Highly resistant Salmonella Newport-MDRAmpC transmitted through the domestic US food supply: a FoodNet case-control study of sporadic Salmonella Newport infections, 2002-2003.

BACKGROUND: A new multidrug-resistant (MDR) strain of Salmonella serotype Newport, Newport-MDRAmpC, has recently emerged. We sought to identify the medical, behavioral, and dietary risk factors for laboratory-confirmed Salmonella Newport infection, including that with Newport-MDRAmpC. METHODS: A 12-month population-based case-control study was conducted during 2002-2003 in 8 sites of the Foodborne Diseases Active Surveillance Network (FoodNet), with 215 case patients with Salmonella Newport infection and 1154 healthy community control subjects. RESULTS: Case patients with Newport-MDRAmpC infection were more likely than control subjects to have taken an antimicrobial agent to which Newport-MDRAmpC is resistant during the 28 days before the onset of diarrheal illness (odds ratio [OR], 5.0 [95% confidence interval {CI}, 1.6-16]). Case patients with Newport-MDRAmpC infection were also more likely to have eaten uncooked ground beef (OR, 7.8 [95% CI, 1.4-44]) or runny scrambled eggs or omelets prepared in the home (OR, 4.9 [95% CI, 1.3-19]) during the 5 days before the onset of illness. International travel was not a risk factor for Newport-MDRAmpC infection but was a strong risk factor for pansusceptible Salmonella Newport infection (OR, 7.1 [95% CI, 2.0-24]). Case patients with pansusceptible infection were also more likely to have a frog or lizard in their household (OR, 2.9 [95% CI, 1.1-7.7]). CONCLUSIONS: Newport-MDRAmpC infection is acquired through the US food supply, most likely from bovine and, perhaps, poultry sources, particularly among persons already taking antimicrobial agents.

Animals↗
PubMed2005

Penetration of Salmonella enteritidis and Salmonella heidelberg into egg yolks in an in vitro contamination model.

Eggs that harbor Salmonella in their edible contents pose a significant risk of transmitting disease to consumers. Although Salmonella deposition inside yolks does not usually occur at a high frequency in naturally contaminated eggs, bacterial penetration through the vitelline membrane could lead to rapid and extensive multiplication in the nutrient-rich yolk contents. The present study used an in vitro egg contamination model to assess the ability of Salmonella strains to penetrate the vitelline membrane and multiply inside yolks. An S. enteritidis strain and 2 Salmonella heidelberg strains, initially inoculated onto the outside of the vitelline membrane, were able to enter the yolk contents (at frequencies ranging from 10 to 25% of experimentally contaminated eggs) during 24 h of incubation at 30 degrees C. Variants of these parent strains, obtained by in vivo passage into eggs laid by infected hens, penetrated the yolk membrane at significantly higher frequencies. These results demonstrate that pathogens such as S. enteritidis and S. heidelberg can penetrate into and begin to multiply inside the yolks of contaminated eggs during the first day of storage at warm temperatures.

Animals↗
PubMed1991

The 106-kilobase plasmid of Salmonella braenderup and the 100-kilobase plasmid of Salmonella typhimurium are not necessary for the pathogenicity in experimental models.

Among 1.041 clinical isolates (77 serovars) of Salmonella which had been derived from cases with acute enterocolitis, 601 (58%) contained one or more plasmids. Large serovar-specific plasmids were seen in 95 of 307 isolates (31%) of Salmonella typhimurium, in 34 of 34 isolates (100%) of Salmonella enteritidis and in 36 of 38 isolates (94.7%) of Salmonella braenderup: the sizes of which were 100, 60 and 106 kilobases (kb), respectively. In order to determine the role of these plasmids in pathogenicity for enterocolitis, the plasmids were eliminated from some strains of S. braenderup and S. typhimurium and the pathogenicity of the plasmid-less strains was compared with that of the parent strains by invasiveness to HeLa cells, fluid accumulation in the rabbit ligated ileal loop, lesion of mucosal tissue and the Sereny test. The virulence of all the plasmid-less strains was as strong as that of the plasmid-bearing strains in these pathogenicity assay systems. We therefore concluded that the 106-kb plasmid of S. braenderup and the 100-kb plasmid of S. typhimurium are not necessary for their pathogenicity in the experimental models: invasiveness to HeLa cells, fluid accumulation in the rabbit ligated ileal loop, and Sereny test.

Animals↗
PubMed1998

Application of random amplified polymorphic DNA analysis to differentiate strains of Salmonella typhi and other Salmonella species.

A Random Amplified Polymorphic DNA (RAPD) fingerprinting method was developed to differentiate isolates of Salmonella serotype typhi (S. typhi) and other Salmonella isolates. A panel of five primers was used to examine 63 isolates of Salm. typhi, including 56 strains isolated in Taiwan and seven strains obtained abroad. Twenty-one RAPD types were revealed using the RAPD fingerprinting method. An RAPD with primer 6032 yielded a polymorphism in a 350 bp fragment that differentiated the attenuated vaccine strain Salm. typhi Ty21a from the rest of the Salm. typhi strains. Strains of Salm. typhi were divided into five types with primer D14307. Primer D14307 also proved capable of discrimination among 65 other Salmonella isolates representing 42 different serotypes. The bacterial DNA used in this RAPD protocol was obtained using a commercially available DNA extraction kit (GeneReleaser). The DNA of various strains of Salmonella from this simple extraction procedure could be discriminated within a few hours using the RAPD technique.

Bacterial Typing Techniques↗
PubMed1994

Comparison of the Salmonella-Tek ELISA to culture methods for detection of Salmonella enteritidis in litter and cloacal swabs of poultry.

An enzyme-linked immunosorbent assay (ELISA), using the Salmonella-Tek ELISA test system, was compared with bacteriological isolation for detection of Salmonella enteritidis phage type 4 in litter and cloacal swabs of poultry. Before ELISA, samples were enriched in tetrathionate broth for 18-24 h or in tetrathionate broth (18-24 h), followed by M-broth/novobiocine (4-6 h). The bacteriological isolation procedure included incubation in buffered peptone water and inoculation on modified semisolid Rappaport-Vassiliadis. For detection of S. enteritidis in litter samples, ELISA was more sensitive than bacteriological examination. For cloacal swabs, both techniques were equally sensitive. The two enrichment procedures gave no significant differences in ELISA results. It was concluded that the Salmonella-Tek ELISA test might be a promising test for detection of Salmonella infected poultry flocks, providing that further studies are performed to determine the cut-off value for litter and cloacal swabs of commercial flocks.

Animals↗
PubMed2006

Detection of Salmonella spp. in stray dogs in Bursa Province, Turkey: first isolation of Salmonella Corvallis from dogs.

The prevalence and antimicrobial sensitivity of Salmonella spp. in stray dogs in Bursa province, Turkey were determined from a total of 82 faecal samples by bacteriological methods. Of the dogs, nine (11%) were positive for salmonellae-carrying. All Salmonella isolates were serotyped as S. Corvallis, and were sensitive to amoxycillin/clavulanic acid, ampicillin, cephalothin, chloramphenicol, enrofloxacin, gentamicin, kanamycin, nalidixic acid, neomycin, oxytetracycline and trimethoprim/sulphamethoxazole, while 66.7% of them were resistant to streptomycin. It was concluded that stray dogs could be an important carrier of salmonellae as well as a source of human salmonellosis in Turkey. Additional studies are needed to clarify the epidemiological relationship between S. Corvallis isolated from dogs and humans with regard to public health. This is the first report on the isolation of S. Corvallis from dogs in the country.

Animals↗
PubMed1996

The Salmonella typhimurium flgM gene, which encodes a negative regulator of flagella synthesis and is involved in virulence, is present and functional in other Salmonella species.

FlgM inhibits the flagella-specific sigma factor FliA and is involved in the mouse-virulence of Salmonella typhimurium. In recent experiments, we observed that: (i) a flgM gene that could function to negatively regulate flagella synthesis was present in a variety of salmonellae; and (ii) the flgM gene derived from Salmonella species that are not normally virulent in mice could complement the S. typhimurium flgM mutant for virulence. Our results suggest that a functional flgM has been retained in most, and perhaps all, Salmonella species, regardless of the motility or virulence phenotype of the strain.

Animals↗
PubMed1979

Artificial Salmonella vaccines: O-antigenic oligosaccharide-protein conjugates induce protection against infection with Salmonella typhimurium.

Outbred mice were vaccinated with various artificial Salmonella vaccines and subsequently challenged intraperitoneally with graded doses of virulent Salmonella typhimurium. The Salmonella vaccines used were: (i) octasaccharide, obtained by hydrolysis of the O-antigenic polysaccharide chain of S. typhimurium strain SH 4809 with phage P22-associated endo-rhamnosidase and covalently linked to either diphtheria toxin or edestine; (ii) purified outer membrane proteins (porins) from S. typhimurium; and (iii) octasaccharide covalently linked to porins. All vaccines induced significant protection against experimental infection of mice with S. typhimurium. However, vaccination with the octasaccharide-porin conjugate resulted in better protection than that obtained by vaccination with octasaccharide or porin vaccines separately. Rabbit antibodies raised against the different vaccines were also passively administered intravenously to mice. Such mice were protected against challenge with virulent S. typhimurium by antibodies specific for the S. typhimurium O-antigen or for the porins. Thus, active immunization with more than one surface component of Salmonella bacteria improved the efficacy of the vaccine. The data from the passive immunization experiments also emphasized the role of humoral immunity for protection against S. typhimurium infection.

Animals↗
PubMed1985

Passage of Salmonella enteritidis and Salmonella thompson through chick ileocecal mucosa.

The passage of Salmonella enteritidis and S. thompson across the cecal mucosa has been visualized in an electron microscope study with the freshly hatched chick as a model. The uptake of salmonellae by macrophages took place in the cecal lumen; the macrophages became abnormal and often ruptured to release organisms back into the lumen. The entry of bacteria into the epithelial cells was associated with a series of pathological changes, beginning with the appearance of active Golgi apparatus and the production of a variety of lysosomal vesicles. Salmonellae became sequestered within lysosomes but were unaffected by the presence of hydrolytic enzyme. Epithelial cell death was related to particularly large numbers of bacteria. Fragments of invaded epithelial cells, especially those undergoing cell death, contributed to the cytoplasmic debris and released further salmonellae into the lumen. Bacteria were never observed in large numbers below the basement membrane, and there was no significant pathology in the lamina propria tissue. Wandering cells, identified as macrophages and containing the bacteria, were observed spanning the epithelial and lamina propria regions through breaks in the basement membrane. It is suggested that the passage of bacteria from the epithelium to the lamina propria is primarily the result of capture and transport within host macrophages.

Animals↗
PubMed1990

Correlation between the presence of sequences homologous to the vir region of Salmonella dublin plasmid pSDL2 and the virulence of twenty-two Salmonella serotypes in mice.

Large plasmids encoding important virulence properties have been found in several Salmonella serotypes. We have studied the relationship between the presence of a highly conserved 4-kilobase (kb) EcoRI fragment from the plasmid virulence region and pathogenicity for mice of 53 isolates representing 22 serotypes of Salmonella. Only strains possessing the homologous 4-kb region were virulent for mice. In addition, we transferred the virulence plasmid from S. dublin into nine different serotypes, including S. typhi and S. paratyphi A, that lack a native virulence plasmid. Only S. heidelberg and S. newport were rendered mouse virulent by the introduction of the S. dublin plasmid. This study demonstrates that plasmid-mediated virulence sequences are required for Salmonella virulence in mice, but many strains, including the agents of human typhoid fever, also lack chromosomal genes necessary to produce lethal systemic disease in mice. Since all the major Salmonella strains that are host-adapted to animals carry virulence plasmids, it appears that these plasmids are important in mediating systemic infection in animals and may contribute to septicemic, nontyphoid salmonellosis in humans.

Animals↗
PubMed1992

Specific lung mucosal and systemic immune responses after oral immunization of mice with Salmonella typhimurium aroA, Salmonella typhi Ty21a, and invasive Escherichia coli expressing recombinant pertussis toxin S1 subunit.

Pertussis toxin (PT) is considered an essential protective component for incorporation into new generation vaccines against Bordetella pertussis, the causative agent of whooping cough. Traditionally, antipertussis vaccination has employed an intramuscular route. An alternative to this approach is to stimulate mucosal and systemic immune responses by oral immunization with live vaccine carrier strains of Salmonella spp. or Escherichia coli. Recombinant S1 subunit of pertussis toxin was expressed in the attenuated aroA mutant of Salmonella typhimurium, SL3261, in the human typhoid vaccine strain Salmonella typhi Ty21a, and in E. coli CAG629 containing the Shigella flexneri plasmid pWR110, which encodes bacterial invasiveness of epithelial cells. Expression of recombinant PT S1 subunit (rPT-S1) did not affect in vitro invasiveness of the tested strains, which retained the ability to adhere to and invade the embryonic human intestinal cell line HI-407. Following oral immunization of mice with the live vaccine strains expressing rPT-S1, immunoglobulin G (IgG), IgA, and IgM responses were monitored. IgG specific to PT was detected in serum samples of mice, while IgG and IgA specific to PT were detected in lung washes after oral immunization with living Salmonella spp. or E. coli (pWR110) expressing rPT-S1. Utilization of live oral vaccines expressing B. pertussis antigens, which stimulate both a systemic and lung mucosal response, may provide an attractive alternative to purified component vaccines against whooping cough.

Administration, Oral↗
PubMed1985

High-molecular-weight components in lipopolysaccharides of Salmonella typhimurium, Salmonella minnesota, and Escherichia coli.

Lipopolysaccharide from smooth strains of Salmonella typhimurium, Salmonella minnesota, and Escherichia coli O111:B4, O55:B5, and O127:B8 was fractionated by gel filtration chromatography. All lipopolysaccharide samples separated into three major populations. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the fractions from S. typhimurium and S. minnesota indicated that the three peaks were made up of molecules with average O-antigen lengths of (i) 70 or more repeat units, (ii) 30 and 20 repeats units in the samples from S. typhimurium and S. minnesota, respectively, and (iii) 1 repeat unit. In contrast to the Salmonella samples, peak 1 from the E. coli samples was not detected on polyacrylamide gels and lacked detectable phosphate. This high-molecular-weight material had a sugar composition similar to that of O-antigen and was tentatively identified as capsular polysaccharide. Peaks 2 and 3 of the E. coli samples were analogous to those of the Salmonella isolates, containing lipopolysaccharide molecules with averages of 18 and 1 O-antigen repeat units, respectively. These lipopolysaccharide molecules did not completely dissociate during electrophoresis, and multimers were detected as distinct, anomalous, slow-migrating bands. Increasing the concentration of sodium dodecyl sulfate in the gels resulted in the dissociation of these multimers.

Antigens, Bacterial↗
PubMed1990

Molecular cloning and physical and functional characterization of the Salmonella typhimurium and Salmonella typhi galactose utilization operons.

The chromosomally encoded galactose utilization (gal) operons of Salmonella typhimurium and S. typhi were each cloned on similar 5.5-kilobase HindIII fragments into pBR322 and were identified by complementation of Gal- Escherichia coli strains. Restriction endonuclease analyses indicated that these Salmonellae operons share considerable homology, but some heterogeneities in restriction sites were observed. Subcloning and exonuclease mapping experiments showed that both operons have the same genetic organization as that established for the E. coli gal operon (i.e., 5' end, promoter, epimerase, transferase, kinase, and 3' end). Two gal operator regions (oE and oI) of S. typhimurium, identified by repressor titration in an E. coli superrepressor [galR(Sup)] mutant, were sequenced and found to flank the promoter region. This promoter region is identical to the -10 and -35 regions of the E. coli gal operon. Minicell studies demonstrated that the three gal structural genes of S. typhimurium encode separate polypeptides of 39 kilodaltons (kDa) (epimerase, 337 amino acids [aa's]), 41 kDa (transferase, 348 aa's), and 43 kDa (kinase, 380 aa's). Despite functional and organizational similarities, DNA sequence analysis revealed that the S. typhimurium gal genes show less than 70% homology to the E. coli gal operon. Because of codon degeneracy, the deduced amino acid sequences of these polypeptides are highly conserved (greater than 90% homology) as compared with those of the E. coli gal enzymes. These studies have defined basic genetic parameters of the gal genes of two medically important Salmonella species, and our findings support the hypothesized divergent evolution of E. coli and Salmonella spp. from a common ancestral parent bacterium.

Amino Acid Sequence↗
PubMed1984

Recovery of Salmonella group B from blood and Salmonella group C2 from feces and serological evidence of dual infection in one patient.

A patient with a dual Salmonella infection is described. Salmonella group B was recovered from three blood culture sets but was not detected in seven stool cultures. Salmonella group C2 was isolated from three of seven stool cultures but was not recovered from blood cultures. Specific, non-cross-reactive antibodies to Salmonella groups B and C2 were detected in the sera of the patient by passive hemagglutination assays.

Adult↗
PubMed1998

Performance of the Dynabeads anti-Salmonella system in the detection of Salmonella species in foods, animal feeds, and environmental samples.

The immunomagnetic separation Dynabeads anti-Salmonella technique was evaluated for the detection of Salmonella species in a variety of foods, feeds, and environmental samples. Salmonella cells in preenrichment broths were captured using the Dynabeads anti-Salmonella system and were then washed and plated on indicator media. A total of 308 naturally contaminated samples were analyzed, including 46 cheese and egg products, 183 animal feeds, and 79 environmental swabs. The results of the Dynabeads method gave 100% correlation with the results of the standard culture technique used for foods and the modified semisolid Rappaport-Vassiliadis method used for feeds and environmental samples.

Animal Feed↗
PubMed2003

Inactivation by ionizing radiation of Salmonella enteritidis, Salmonella infantis, and Vibrio parahaemolyticus in oysters (Crassostrea brasiliana).

Irradiation is considered one of the most efficient technological processes for the reduction of microorganisms in food. It can be used to improve the safety of food products, and to extend their shelf lives. Oysters are considered one of the most important vehicles for pathogenic bacteria because of their feeding characteristics. The aim of this study was to evaluate the influence of a gamma radiation process on high levels of Salmonella Enteritidis, Salmonella Infantis, and Vibrio parahaemolyticus incorporated by oysters (Crassostrea brasiliana), as well as the effects of the process on the survival of the oysters and on their sensory attributes. The oysters were exposed to gamma radiation (60Co) in doses ranging from 0.5 to 3.0 kGy. A dose of 3.0 kGy was generally sufficient to reduce the level of Salmonella serotypes by 5 to 6 log10 units. A dose of 1.0 kGy was sufficient to produce a 6-log10 reduction in the level of V. parahaemolyticus. The highest irradiation dose did not kill the oysters or affect their sensory attributes. Hence, a dose of 3.0 kGy can be considered effective in inactivating Salmonella and V. parahaemolyticus in oysters without changing their odor, flavor, or appearance.

Animals↗