Sensitive and specific detection of E. coli, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium in milk by microchip electrophoresis combined with multiplex PCR amplification

2020 ◽  
Vol 157 ◽  
pp. 104876
Author(s):  
Yan Zhang ◽  
Xianzhi Hu ◽  
Qingjiang Wang
Keyword(s):  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Pcr Amplification ◽  
Microchip Electrophoresis ◽  
Specific Detection ◽  
E Coli ◽  
Serovar Typhimurium

Detection of Escherichia coli O157, Salmonella enterica Serovar Typhimurium, and Staphylococcal Enterotoxin B in a Single Sample Using Enzymatic Bio-Nanotransduction

2007 ◽  
Vol 70 (4) ◽  
pp. 841-850 ◽  
Author(s):  
JOSH R. BRANEN ◽  
MARTHA J. HASS ◽  
ERIN R. DOUTHIT ◽  
WUSI C. MAKI ◽  
A. LARRY BRANEN
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Staphylococcal Enterotoxin B ◽  
Escherichia Coli O157 ◽  
Dna Templates ◽  
E Coli ◽  
Serovar Typhimurium ◽  
Biological Recognition ◽  
Enterotoxin B

Enzymatic bio-nanotransduction is a biological detection scheme based on the production of nucleic acid nano-signals (RNA) in response to specific biological recognition events. In this study, we applied an enzymatic bio-nanotransduction system to the detection of important food-related pathogens and a toxin. Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and staphylococcal enterotoxin B (SEB) were chosen because of the implications of these targets to food safety. Primary antibodies to each of the targets were used to functionalize magnetic beads and produce biological recognition elements (antibodies) conjugated to nano-signal–producing DNA templates. Immunomagnetic capture that was followed by in vitro transcription of DNA templates bound to target molecules produced RNA nano-signals specific for every target in the sample. Discrimination of RNA nano-signals with a standard enzyme-linked oligonucleotide fluorescence assay provided a correlation between nano-signal profiles and target concentrations. The estimated limit of detection was 2.4 × 103 CFU/ml for E. coli O157:H7, 1.9 × 104 CFU/ml for S. enterica serovar Typhimurium, and 0.11 ng/ml for SEB with multianalyte detection in buffer. Low levels of one target were also detected in the presence of interference from high levels of the other targets. Finally, targets were detected in milk, and detection was improved for E. coli O157 by heat treatment of the milk.

scholarly journals Salmonella enterica Serovar Typhimurium and Escherichia coli Survival in Estuarine Bank Sediments

2018 ◽  
Vol 15 (11) ◽  
pp. 2597 ◽  
Author(s):  
Mahbubul Siddiqee ◽  
Rebekah Henry ◽  
Rebecca Coulthard ◽  
Christelle Schang ◽  
Richard Williamson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Growth Patterns ◽  
Estuarine Sediments ◽  
Human Pathogens ◽  
Indicator Organisms ◽  
Fecal Indicator ◽  
E Coli ◽  
Serovar Typhimurium

Estuarine bank sediments have the potential to support the survival and growth of fecal indicator organisms, including Escherichia coli. However, survival of fecal pathogens in estuarine sediments is not well researched and therefore remains a significant knowledge gap regarding public health risks in estuaries. In this study, simultaneous survival of Escherichia coli and a fecal pathogen, Salmonella enterica serovar Typhimurium, was studied for 21 days in estuarine bank sediment microcosms. Observed growth patterns for both organisms were comparable under four simulated scenarios; for continuous-desiccation, extended-desiccation, periodic-inundation, and continuous-inundation systems, logarithmic decay coefficients were 1.54/day, 1.51/day, 0.14/day, and 0.20/day, respectively, for E. coli, and 1.72/day, 1.64/day, 0.21/day, and 0.24/day for S. Typhimurium. Re-wetting of continuous-desiccated systems resulted in potential re-growth, suggesting survival under moisture-limited conditions. Key findings from this study include: (i) Bank sediments can potentially support human pathogens (S. Typhimurium), (ii) inundation levels influence the survival of fecal bacteria in estuarine bank sediments, and (iii) comparable survival rates of S. Typhimurium and E. coli implies the latter could be a reliable fecal indicator in urban estuaries. The results from this study will help select suitable monitoring and management strategies for safer recreational activities in urban estuaries.

scholarly journals The Multi-Copper-Ion Oxidase CueO of Salmonella enterica Serovar Typhimurium Is Required for Systemic Virulence

2010 ◽  
Vol 78 (5) ◽  
pp. 2312-2319 ◽  
Author(s):  
Maud E. S. Achard ◽  
Jai J. Tree ◽  
James A. Holden ◽  
Kim R. Simpfendorfer ◽  
Odilia L. C. Wijburg ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Copper Ions ◽  
Copper Ion ◽  
Mesenteric Lymph Nodes ◽  
E Coli ◽  
Significant Difference ◽  
Serovar Typhimurium ◽  
Branched Chain ◽  
Periplasmic Enzyme

ABSTRACT Salmonella enterica serovar Typhimurium possesses a multi-copper-ion oxidase (multicopper oxidase), CueO (also known as CuiD), a periplasmic enzyme known to be required for resistance to copper ions. CueO from S. Typhimurium was expressed as a recombinant protein in Escherichia coli, and the purified protein exhibited a high cuprous oxidase activity. We have characterized an S. Typhimurium cueO mutant and confirmed that it is more sensitive to copper ions. Using a murine model of infection, it was observed that the cueO mutant was significantly attenuated, as indicated by reduced recovery of bacteria from liver and spleen, although there was no significant difference in recovery from Peyer's patches and mesenteric lymph nodes. However, the intracellular survival of the cueO mutant in unprimed or gamma-interferon-primed murine macrophages was not statistically different from that of wild-type Salmonella, suggesting that additional host factors are involved in clearance of the cueO mutant. Unlike a cueO mutant from E. coli, the S. Typhimurium cueO mutant did not show greater sensitivity to hydrogen peroxide and its sensitivity to copper ions was not affected by siderophores. Similarly, the S. Typhimurium cueO mutant was not rescued from copper ion toxicity by addition of the branched-chain amino acids and leucine.

Influence of the Treatment of Listeria monocytogenes and Salmonella enterica Serovar Typhimurium with Citral on the Efficacy of Various Antibiotics

2014 ◽  
Vol 11 (4) ◽  
pp. 265-271 ◽  
Author(s):  
Surama F. Zanini ◽  
Angela B. Silva-Angulo ◽  
Amauri Rosenthal ◽  
Dolores Rodrigo Aliaga ◽  
Antonio Martínez
Keyword(s):  
Listeria Monocytogenes ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Serovar Typhimurium

scholarly journals Biofilm Formation by Salmonella enterica Serovar Typhimurium and Escherichia coli on Epithelial Cells following Mixed Inoculations

2005 ◽  
Vol 73 (8) ◽  
pp. 5198-5203 ◽  
Author(s):  
Cristina L. C. Esteves ◽  
Bradley D. Jones ◽  
Steven Clegg
Keyword(s):  
Escherichia Coli ◽  
Epithelial Cells ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Salmonella Enterica Serovar Typhimurium ◽  
E Coli ◽  
Serovar Typhimurium

ABSTRACT Biofilms were formed by inoculations of Salmonella enterica serovar Typhimurium and Escherichia coli on HEp-2 cells. Inoculations of S. enterica serovar Typhimurium and E. coli resulted in the formation of an extensive biofilm of S. enterica serovar Typhimurium. In experiments where an E. coli biofilm was first formed followed by challenge with S. enterica serovar Typhimurium, there was significant biofilm formation by S. enterica serovar Typhimurium. The results of this study indicate that S. enterica serovar Typhimurium can outgrow E. coli in heterologous infections and displace E. coli when it forms a biofilm on HEp-2 cells.

scholarly journals Aspartic Peptide Hydrolases in Salmonella enterica Serovar Typhimurium

2001 ◽  
Vol 183 (10) ◽  
pp. 3089-3097 ◽  
Author(s):  
Rachel A. Larsen ◽  
Tina M. Knox ◽  
Charles G. Miller
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Gene Encoding ◽  
E Coli ◽  
Mutant Strains ◽  
Serovar Typhimurium ◽  
Measurable Rate ◽  
Mature Enzyme ◽  
Aminopeptidase A ◽  
Peptide Hydrolases

ABSTRACT Two well-characterized enzymes in Salmonella entericaserovar Typhimurium and Escherichia coli are able to hydrolyze N-terminal aspartyl (Asp) dipeptides: peptidase B, a broad-specificity aminopeptidase, and peptidase E, an Asp-specific dipeptidase. A serovar Typhimurium strain lacking both of these enzymes, however, can still utilize most N-terminal Asp dipeptides as sources of amino acids, and extracts of such a strain contain additional enzymatic activities able to hydrolyze Asp dipeptides. Here we report two such activities from extracts of pepB pepEmutant strains of serovar Typhimurium identified by their ability to hydrolyze Asp-Leu. Although each of these activities hydrolyzes Asp-Leu at a measurable rate, the preferred substrates for both are N-terminal isoAsp peptides. One of the activities is a previously characterized isoAsp dipeptidase from E. coli, the product of theiadA gene. The other is the product of the serovar Typhimurium homolog of E. coli ybiK, a gene of previously unknown function. This gene product is a member of the N-terminal nucleophile structural family of amidohydrolases. Like most other members of this family, the mature enzyme is generated from a precursor protein by proteolytic cleavage and the active enzyme is a heterotetramer. Based on its ability to hydrolyze an N-terminal isoAsp tripeptide as well as isoAsp dipeptides, the enzyme appears to be an isoAsp aminopeptidase, and we propose that the gene encoding it be designated iaaA (isoAsp aminopeptidase). A strain lacking both IadA and IaaA in addition to peptidase B and peptidase E has been constructed. This strain utilizes Asp-Leu as a leucine source, and extracts of this strain contain at least one additional, as-yet-uncharacterized, peptidase able to cleave Asp dipeptides.

scholarly journals Arginine-Dependent Acid Resistance in Salmonella enterica Serovar Typhimurium

2006 ◽  
Vol 188 (15) ◽  
pp. 5650-5653 ◽  
Author(s):  
Jasper Kieboom ◽  
Tjakko Abee
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Acid Resistance ◽  
Arginine Decarboxylase ◽  
Acidic Ph ◽  
E Coli ◽  
Serovar Typhimurium ◽  
Microaerobic Conditions ◽  
Acid Challenge

ABSTRACT Salmonella enterica serovar Typhimurium does not survive a pH 2.5 acid challenge under conditions similar to those used for Escherichia coli (J. W. Foster, Nat. Rev. Microbiol. 2:898-907, 2004). Here, we provide evidence that S. enterica serovar Typhimurium can display arginine-dependent acid resistance (AR) provided the cells are grown under anoxic conditions and not under the microaerobic conditions used for assessment of AR in E. coli. The role of the arginine decarboxylase pathway in Salmonella AR was shown by the loss of AR in mutants lacking adiA, which encodes arginine decarboxylase; adiC, which encodes the arginine-agmatine antiporter; or adiY, which encodes an AraC-like regulator. Transcription of adiA and adiC was found to be dependent on AdiY, anaerobiosis, and acidic pH.

scholarly journals Effect of Inactivation of degS on Salmonella enterica Serovar Typhimurium In Vitro and In Vivo

2005 ◽  
Vol 73 (1) ◽  
pp. 459-463 ◽  
Author(s):  
Gary Rowley ◽  
Andrew Stevenson ◽  
Jan Kormanec ◽  
Mark Roberts
Keyword(s):  
Sigma Factor ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Wild Type ◽  
Alternative Pathways ◽  
E Coli ◽  
Mammalian Tissues ◽  
Serovar Typhimurium

ABSTRACT The alternative sigma factor (RpoE σE) enables Salmonella enterica serovar Typhimurium to adapt to stressful conditions, such as oxidative stress, nutrient deprivation, and growth in mammalian tissues. Infection of mice by Salmonella serovar Typhimurium also requires σE. In Escherichia coli, activation of the σE pathway is dependent on proteolysis of the anti-sigma factor RseA and is initiated by DegS. DegS is also important in order for E. coli to cause extraintestinal infection in mice. We constructed a degS mutant of the serovar Typhimurium strain SL1344 and compared its behavior in vitro and in vivo with those of its wild-type (WT) parent and an isogenic rpoE mutant. Unlike E. coli degS strains, the Salmonella serovar Typhimurium degS strain grew as well as the WT strain at 42°C. The degS mutant survived very poorly in murine macrophages in vitro and was highly attenuated compared with the WT strain for both the oral and parenteral routes of infection in mice. However, the degS mutant was not as attenuated as the serovar Typhimurium rpoE mutant: 100- to 1,000-fold more degS bacteria than rpoE bacteria were present in the livers and spleens of mice 24 h after intraperitoneal challenge. In most assays, the rpoE mutant was more severely affected than the degS mutant and a σE-dependent reporter gene was more active in the degS mutant than the rpoE strain. These findings indicate that degS is important for activation of the σE pathway in serovar Typhimurium but that alternative pathways for σE activation probably exist.

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