scholarly journals Effect of Frequency and Waveform on Inactivation of Escherichia coli O157:H7 and Salmonella enterica Serovar Typhimurium in Salsa by Ohmic Heating

2012 ◽  
Vol 79 (1) ◽  
pp. 10-17 ◽  
Author(s):  
Su-Yeon Lee ◽  
Sangryeol Ryu ◽  
Dong-Hyun Kang
Keyword(s):  
Escherichia Coli ◽  
Electrical Conductivity ◽  
Heating Rate ◽  
Salmonella Enterica ◽  
Treatment Time ◽  
Ohmic Heating ◽  
Content Type ◽  
Significant Difference ◽  
Serovar Typhimurium ◽  
The Impact

ABSTRACT The effect of frequency of alternating current during ohmic heating on electrode corrosion, heating rate, inactivation of food-borne pathogens, and quality of salsa was investigated. The impact of waveform on heating rate was also investigated. Salsa was treated with various frequencies (60 Hz to 20 kHz) and waveforms (sine, square, and sawtooth) at a constant electric field strength of 12.5 V/cm. Electrode corrosion did not occur when the frequency exceeded 1 kHz. The heating rate of the sample was dependent on frequency up to 500 Hz, but there was no significant difference ( P > 0.05) in the heating rate when the frequency was increased above 1 kHz. The electrical conductivity of the sample increased with a rise in the frequency. At a frequency of 60 Hz, the square wave produced a lower heating rate than that of sine and sawtooth waves. The heating rate between waveforms was not significantly ( P > 0.05) different when the frequency was >500 Hz. As the frequency increased, the treatment time required to reduce Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium to below the detection limit (1 log CFU/g) decreased without affecting product quality. These results suggest that ohmic heating can be effectively used to pasteurize salsa and that the effect of inactivation is dependent on frequency and electrical conductivity rather than waveform.

scholarly journals Effect of Electropermeabilization by Ohmic Heating for Inactivation of Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Buffered Peptone Water and Apple Juice

2013 ◽  
Vol 79 (23) ◽  
pp. 7122-7129 ◽  
Author(s):  
Il-Kyu Park ◽  
Dong-Hyun Kang
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Electric Field ◽  
Salmonella Enterica ◽  
Apple Juice ◽  
Ohmic Heating ◽  
Conventional Heating ◽  
Content Type ◽  
Serovar Typhimurium ◽  
Peptone Water

ABSTRACTThe effect of electric field-induced ohmic heating for inactivation ofEscherichia coliO157:H7,Salmonella entericaserovar Typhimurium, andListeria monocytogenesin buffered peptone water (BPW) (pH 7.2) and apple juice (pH 3.5; 11.8 °Brix) was investigated in this study. BPW and apple juice were treated at different temperatures (55°C, 58°C, and 60°C) and for different times (0, 10, 20, 25, and 30 s) by ohmic heating compared with conventional heating. The electric field strength was fixed at 30 V/cm and 60 V/cm for BPW and apple juice, respectively. Bacterial reduction resulting from ohmic heating was significantly different (P< 0.05) from that resulting from conventional heating at 58°C and 60°C in BPW and at 55°C, 58°C, and 60°C in apple juice for intervals of 0, 10, 20, 25, and 30 s. These results show that electric field-induced ohmic heating led to additional bacterial inactivation at sublethal temperatures. Transmission electron microscopy (TEM) observations and the propidium iodide (PI) uptake test were conducted after treatment at 60°C for 0, 10, 20, 25 and 30 s in BPW to observe the effects on cell permeability due to electroporation-caused cell damage. PI values when ohmic and conventional heating were compared were significantly different (P< 0.05), and these differences increased with increasing levels of inactivation of three food-borne pathogens. These results demonstrate that ohmic heating can more effectively reduce bacterial populations at reduced temperatures and shorter time intervals, especially in acidic fruit juices such as apple juice. Therefore, loss of quality can be minimized in a pasteurization process incorporating ohmic heating.

scholarly journals Display of Recombinant Proteins on Bacterial Outer Membrane Vesicles by Using Protein Ligation

2018 ◽  
Vol 84 (8) ◽  
pp. e02567-17 ◽  
Author(s):  
H. Bart van den Berg van Saparoea ◽  
Diane Houben ◽  
Marien I. de Jonge ◽  
Wouter S. P. Jong ◽  
Joen Luirink
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Salmonella Enterica ◽  
Membrane Vesicles ◽  
Therapeutic Agents ◽  
Outer Membrane Vesicles ◽  
High Density ◽  
Content Type ◽  
Protein Ligation ◽  
Serovar Typhimurium

ABSTRACT The Escherichia coli virulence factor hemoglobin protease (Hbp) has been engineered into a surface display system that can be expressed to high density on live E. coli and Salmonella enterica serovar Typhimurium cells or derived outer membrane vesicles (OMVs). Multiple antigenic sequences can be genetically fused into the Hbp core structure for optimal exposure to the immune system. Although the Hbp display platform is relatively tolerant, increasing the number, size, and complexity of integrated sequences generally lowers the expression of the fused constructs and limits the density of display. This is due to the intricate mechanism of Hbp secretion across the outer membrane and the efficient quality control of translocation-incompetent chimeric Hbp molecules in the periplasm. To address this shortcoming, we explored the coupling of purified proteins to the Hbp carrier after its translocation across the outer membrane using the recently developed SpyTag/SpyCatcher protein ligation system. As expected, fusion of the small SpyTag to Hbp did not hamper display on OMVs. Subsequent addition of purified proteins fused to the SpyCatcher domain resulted in efficient covalent coupling to Hbp-SpyTag. Using in addition the orthogonal SnoopTag/SnoopCatcher system, multiple antigen modules could be coupled to Hbp in a sequential ligation strategy. Not only antigens proved suitable for Spy-mediated ligation but also nanobodies. Addition of this functionality to the platform might allow the targeting of live bacterial or OMV vaccines to certain tissues or immune cells to tailor immune responses.IMPORTANCE Outer membrane vesicles (OMVs) derived from Gram-negative bacteria attract increasing interest in the development of vaccines and therapeutic agents. We aim to construct a semisynthetic OMV platform for recombinant antigen presentation on OMVs derived from attenuated Salmonella enterica serovar Typhimurium cells displaying an adapted Escherichia coli autotransporter, Hbp, at the surface. Although this autotransporter accepts substantial modifications, its capacity with respect to the number, size, and structural complexity of the antigens genetically fused to the Hbp carrier is restricted. Here we describe the application of SpyCatcher/SpyTag protein ligation technology to enzymatically link antigens to Hbp present at high density in OMVs. Protein ligation was apparently unobstructed by the membrane environment and allowed a high surface density of coupled antigens, a property we have shown to be important for vaccine efficacy. The OMV coupling procedure appears versatile and robust, allowing fast production of experimental vaccines and therapeutic agents through a modular plug-and-display procedure.

scholarly journals An Attenuated Salmonella enterica Serovar Typhimurium Strain and Galacto-Oligosaccharides Accelerate Clearance of Salmonella Infections in Poultry through Modifications to the Gut Microbiome

2017 ◽  
Vol 84 (5) ◽  
Author(s):  
M. Andrea Azcarate-Peril ◽  
Natasha Butz ◽  
Maria Belen Cadenas ◽  
Matthew Koci ◽  
Anne Ballou ◽  
...  
Keyword(s):  
United States ◽  
Gut Microbiota ◽  
Gut Microbiome ◽  
Salmonella Enterica ◽  
The United States ◽  
Wild Type ◽  
Content Type ◽  
Salmonella Infections ◽  
Serovar Typhimurium ◽  
The Impact

ABSTRACT Salmonella is estimated to cause one million foodborne illnesses in the United States every year. Salmonella -contaminated poultry products are one of the major sources of salmonellosis. Given the critical role of the gut microbiota in Salmonella transmission, a manipulation of the chicken intestinal microenvironment could prevent animal colonization by the pathogen. In Salmonella , the global regulator gene fnr ( f umarate n itrate r eduction) regulates anaerobic metabolism and is essential for adapting to the gut environment. This study tested the hypothesis that an attenuated Fnr mutant of Salmonella enterica serovar Typhimurium (attST) or prebiotic galacto-oligosaccharides (GOS) could improve resistance to wild-type Salmonella via modifications to the structure of the chicken gut microbiome. Intestinal samples from a total of 273 animals were collected weekly for 9 weeks to evaluate the impact of attST or prebiotic supplementation on microbial species of the cecum, duodenum, jejunum, and ileum. We next analyzed changes to the gut microbiome induced by challenging the animals with a wild-type Salmonella serovar 4,[5],12:r:− (Nal r ) strain and determined the clearance rate of the virulent strain in the treated and control groups. Both GOS and the attenuated Salmonella strain modified the gut microbiome but elicited alterations of different taxonomic groups. The attST produced significant increases of Alistipes and undefined Lactobacillus , while GOS increased Christensenellaceae and Lactobacillus reuteri . The microbiome structural changes induced by both treatments resulted in a faster clearance after a Salmonella challenge. IMPORTANCE With an average annual incidence of 13.1 cases/100,000 individuals, salmonellosis has been deemed a nationally notifiable condition in the United States by the Centers for Disease Control and Prevention (CDC). Earlier studies demonstrated that Salmonella is transmitted by a subset of animals (supershedders). The supershedder phenotype can be induced by antibiotics, ascertaining an essential role for the gut microbiota in Salmonella transmission. Consequently, modulation of the gut microbiota and modification of the intestinal microenvironment could assist in preventing animal colonization by the pathogen. Our study demonstrated that a manipulation of the chicken gut microbiota by the administration of an attenuated Salmonella strain or prebiotic galacto-oligosaccharides (GOS) can promote resistance to Salmonella colonization via increases of beneficial microorganisms that translate into a less hospitable gut microenvironment.

scholarly journals Whole-Genome Sequence of Salmonella enterica subsp. enterica Serovar Typhimurium Strain WG49 and Escherichia coli Strain WG5 Used in South Africa for Phage Detection in Water Samples

2018 ◽  
Vol 6 (26) ◽  
Author(s):  
L. Bothma ◽  
D. Gonzalez-Ibeas ◽  
C. Mienie ◽  
C. C. Bezuidenhout ◽  
R. Adeleke
Keyword(s):  
Escherichia Coli ◽  
South Africa ◽  
Genome Sequence ◽  
Water Samples ◽  
Salmonella Enterica ◽  
Coli Strain ◽  
Whole Genome Sequence ◽  
Whole Genome ◽  
Content Type ◽  
Serovar Typhimurium

Salmonella enterica subsp. enterica serovar Typhimurium WG49 is widely used for enumeration of F-specific RNA (F-RNA) coliphages in water.

scholarly journals Survival of Host-Associated Bacteroidales Cells and Their Relationship with Enterococcus spp., Campylobacter jejuni, Salmonella enterica Serovar Typhimurium, and Adenovirus in Freshwater Microcosms as Measured by Propidium Monoazide-Quantitative PCR

2011 ◽  
Vol 78 (4) ◽  
pp. 922-932 ◽  
Author(s):  
Sungwoo Bae ◽  
Stefan Wuertz
Keyword(s):  
Quantitative Pcr ◽  
Campylobacter Jejuni ◽  
Salmonella Enterica ◽  
Sunlight Exposure ◽  
Propidium Monoazide ◽  
Content Type ◽  
Significant Difference ◽  
Serovar Typhimurium ◽  
Dark Conditions ◽  
Realistic Information

ABSTRACTThe ideal host-associated genetic fecal marker would be capable of predicting the presence of specific pathogens of concern. Flowthrough freshwater microcosms containing mixed feces and inocula of the pathogensCampylobacter jejuni,Salmonella entericaserovar Typhimurium, and adenovirus were placed at ambient temperature in the presence and absence of diurnal sunlight. The totalEnterococcusDNA increased during the early periods (23 h) under sunlight exposure, even though cultivableEnterococcusand DNA in intact cells, as measured by propidium monoazide (PMA), decreased with first-order kinetics during the entire period. We found a significant difference in the decay of host-associatedBacteroidalescells between sunlight exposure and dark conditions (Pvalue < 0.05), whereas the persistence of host-associatedBacteroidalesDNA was comparable. The 2-log reduction times of adenovirus were 72 h for sunlight exposure and 99 h for dark conditions with similar decay rate constants (Pvalue = 0.13). The persistences of fecalBacteroidalescells andCampylobactercells exposed to sunlight were similar, and host-associatedBacteroidalesDNA and waterborne pathogen DNA were degraded at comparable rates (Pvalues > 0.05). Overall, the ratio of quantitative PCR (qPCR) cycle threshold (CT) values with and without PMA treatment was indicative of the time elapsed since inoculation of the microcosm with (i) fecal material from different animal sources based on host-associatedBacteroidalesand (ii) pure cultures of bacterial pathogens. The use of both PMA-qPCR and qPCR may yield more realistic information about recent sources of fecal contamination and result in improved prediction of waterborne pathogens and assessment of health risk.

scholarly journals The Acyl Homoserine Lactone Receptor, SdiA, of Escherichia coli and Salmonella enterica Serovar Typhimurium Does Not Respond to Indole

2012 ◽  
Vol 78 (15) ◽  
pp. 5424-5431 ◽  
Author(s):  
Anice Sabag-Daigle ◽  
Jitesh A. Soares ◽  
Jenée N. Smith ◽  
Mohamed E. Elmasry ◽  
Brian M. M. Ahmer
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Homoserine Lactone ◽  
Acyl Homoserine Lactone ◽  
Content Type ◽  
E Coli ◽  
Serovar Typhimurium ◽  
High Concentrations

ABSTRACTIn this study, we tested the hypothesis that the SdiA proteins ofEscherichia coliandSalmonella entericaserovar Typhimurium respond to indole. While indole was found to have effects on gene expression and biofilm formation, these effects were notsdiAdependent. However, high concentrations of indole did inhibitN-acyl-l-homoserine lactone (AHL) sensing by SdiA. We conclude that SdiA does not respond to indole but indole can inhibit SdiA activity inE. coliandSalmonella.

scholarly journals Effects of Norspermidine and Spermidine on Biofilm Formation by Potentially Pathogenic Escherichia coli and Salmonella enterica Wild-Type Strains

2015 ◽  
Vol 81 (6) ◽  
pp. 2226-2232 ◽  
Author(s):  
Live L. Nesse ◽  
Kristin Berg ◽  
Lene K. Vestby
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Wild Type ◽  
Content Type ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Serovar Typhimurium ◽  
And Control ◽  
Type Strains

ABSTRACTPolyamines are present in all living cells. In bacteria, polyamines are involved in a variety of functions, including biofilm formation, thus indicating that polyamines may have potential in the control of unwanted biofilm. In the present study, the effects of the polyamines norspermidine and spermidine on biofilms of 10 potentially pathogenic wild-type strains ofEscherichia coliserotype O103:H2,Salmonella entericasubsp.entericaserovar Typhimurium, andS. entericaserovar Agona were investigated. We found that exogenously supplied norspermidine and spermidine did not mediate disassembly of preformed biofilm of any of theE. coliandS. entericastrains. However, the polyamines did affect biofilm production. Interestingly, the two species reacted differently to the polyamines. Both polyamines reduced the amount of biofilm formed byE. colibut tended to increase biofilm formation byS. enterica. Whether the effects observed were due to the polyamines specifically targeting biofilm formation, being toxic for the cells, or maybe a combination of the two, is not known. However, there were no indications that the effect was mediated through binding to exopolysaccharides, as earlier suggested forE. coli. Our results indicate that norspermidine and spermidine do not have potential as inhibitors ofS. entericabiofilm. Furthermore, we found that the commercial polyamines used contributed to the higher pH of the test medium. Failure to acknowledge and control this important phenomenon may lead to misinterpretation of the results.

scholarly journals Chlortetracycline Enhances Tonsil Colonization and Fecal Shedding of Multidrug-ResistantSalmonella entericaSerovar Typhimurium DT104 without Major Alterations to the Porcine Tonsillar and Intestinal Microbiota

2018 ◽  
Vol 85 (4) ◽  
Author(s):  
Devin B. Holman ◽  
Bradley L. Bearson ◽  
Heather K. Allen ◽  
Daniel C. Shippy ◽  
Crystal L. Loving ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
The United States ◽  
Multidrug Resistant ◽  
Foodborne Illness ◽  
Content Type ◽  
Fecal Shedding ◽  
Typhimurium Strain ◽  
Serovar Typhimurium ◽  
Collateral Effects ◽  
The Impact

ABSTRACTSalmonellaspp. are estimated to cause 1.2 million cases of human foodborne illness each year in the United States, and pigs can often be asymptomatically colonized withSalmonellaspp. (>50% of farms). Recent reports state that 18.3% ofSalmonella entericaserovar Typhimurium isolates are resistant to ≥3 antimicrobial classes, and multidrug-resistant (MDR) strains are associated with an increased hospitalization rate and other complications. Chlortetracycline is commonly used in swine production to prevent/treat various diseases; therefore, chlortetracycline treatment of pigs unknowingly colonized with MDRSalmonellamay have collateral effects onSalmonellaspp. (and other gut bacteria). In this study, we determined the effect of in-feed chlortetracycline (400 g/ton) on shedding and colonization of pigs challenged with the MDRS. Typhimurium strain DT104 (n= 11/group). We also assessed the impact on the fecal microbiota over the 12-day experimental period and on the ileum, cecum, and tonsil microbiota at 7 days postinoculation (dpi). In MDRS. Typhimurium-inoculated pigs, chlortetracycline administration significantly increased fecal shedding at 2 dpi (+1.4 log10CFU/g;P < 0.001) and enhanced tonsil colonization (+3.1 log10CFU/g;P < 0.001). There were few major alterations detected in the gut or tonsillar microbiota of pigs treated with MDRS. Typhimurium and/or chlortetracycline. The tonsillar transcriptome was largely unaffected despite increased colonization by MDRS. Typhimurium following inoculation of the chlortetracycline-treated pigs. These results highlight the idea that chlortetracycline administration can enhance shedding and colonization of MDRS. Typhimurium in pigs, which could increase the risk of environmental dissemination of MDRSalmonellastrains.IMPORTANCESalmonellaspp. are an important cause of foodborne illness in North America, and pork products are associated with sporadic cases and outbreaks of human salmonellosis. Isolates ofSalmonellamay be resistant to multiple antibiotics, and infections with multidrug-resistant (MDR)Salmonellaspp. are more difficult to treat, leading to increased hospitalization rates. Swine operations commonly use antimicrobials, such as chlortetracycline, to prevent/treat infections, which may have collateral effects on pig microbial populations. Recently, we demonstrated that chlortetracycline induces the expression of genes associated with pathogenesis and invasion in MDRSalmonella entericaserovar Typhimuriumin vitro. In our current study, we show increased tonsillar colonization and fecal shedding of the MDRS. Typhimurium strain DT104 from pigs administered chlortetracycline. Therefore, pigs unknowingly colonized with multidrug-resistantSalmonellaspp. and receiving chlortetracycline for an unrelated infection may be at a greater risk for disseminating MDRSalmonellaspp. to other pigs and to humans through environmental or pork product contamination.

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