Journal: Microbiology Spectrum

Article Title: Prophylactic Administration of a Bacteriophage Cocktail Is Safe and Effective in Reducing Salmonella enterica Serovar Typhimurium Burden in Vivo

doi: 10.1128/Spectrum.00497-21

Figure Lengend Snippet: SalmoFresh delays the burden of Salmonella after challenge in OMM 12 mice. (A) Experimental design. Groups of 4 to 6 OMM 12 mice were orally administered PBS or SalmoFresh (1 × 10 9 PFU) on the indicated days. On day 2, mice were challenged with ST784 (1 × 10 8 CFU) or a PBS control as indicated. (B) Mice were weighed daily. Shown are the percentages of weight loss compared to starting weights of OMM 12 mice over time. (C and D) The amounts of ST784 (CFU) (C) and phages (PFU) (D) in OMM 12 mouse feces were quantitated daily (day 2 corresponds to 3 h after ST784 challenge). (E and F) Intestinal organs were collected on day 6 and homogenized, and the amounts of ST784 (CFU) (E) and phage (PFU) (F) were quantitated. Statistical analyses are described in Materials and Methods and reported in Table S1.

Article Snippet: S. enterica subsp. enterica serovar Typhimurium strain ST784 was obtained from Intralytix, Inc. (Columbia, MD), and routinely cultured in lysogeny broth (LB), on LB agar, or on Drigalski agar (Bio-Rad, Hercules, CA) at 37°C.

Techniques: Control

Journal: Microbiology Spectrum

Article Title: Prophylactic Administration of a Bacteriophage Cocktail Is Safe and Effective in Reducing Salmonella enterica Serovar Typhimurium Burden in Vivo

doi: 10.1128/Spectrum.00497-21

Figure Lengend Snippet: SalmoFresh reduces disease symptoms associated with the burden of Salmonella in OMM 12 mice

Article Snippet: S. enterica subsp. enterica serovar Typhimurium strain ST784 was obtained from Intralytix, Inc. (Columbia, MD), and routinely cultured in lysogeny broth (LB), on LB agar, or on Drigalski agar (Bio-Rad, Hercules, CA) at 37°C.

Techniques:

Journal: Microbiology Spectrum

Article Title: Prophylactic Administration of a Bacteriophage Cocktail Is Safe and Effective in Reducing Salmonella enterica Serovar Typhimurium Burden in Vivo

doi: 10.1128/Spectrum.00497-21

Figure Lengend Snippet: FOP reduces Salmonella burden in vivo . (A) Experimental design. OMM 12 mice were orally administered PBS or FOP (1 × 10 9 PFU) on the indicated days. On day 2, all mice were challenged with strain ST784 (1 × 10 8 CFU). Stars indicate the time points at which samples were taken for the 16S rRNA gene analysis reported in Fig. 3 . (B) Mice were weighed daily. Shown are the percentages of weight loss compared to starting weights of OMM 12 mice over time. (C and D) The amounts of ST784 (CFU) (C) and phages (PFU) (D) in OMM 12 mouse feces were quantitated daily (day 2 corresponds to 3 h after ST784 challenge). (E and F) Intestinal organs were collected on day 6, and the amounts of ST784 (CFU) (E) and phages (PFU) quantitated. Statistical analyses are described in Materials and Methods and reported in Table S2.

Article Snippet: S. enterica subsp. enterica serovar Typhimurium strain ST784 was obtained from Intralytix, Inc. (Columbia, MD), and routinely cultured in lysogeny broth (LB), on LB agar, or on Drigalski agar (Bio-Rad, Hercules, CA) at 37°C.

Techniques: In Vivo

Journal: Microbiology Spectrum

Article Title: Prophylactic Administration of a Bacteriophage Cocktail Is Safe and Effective in Reducing Salmonella enterica Serovar Typhimurium Burden in Vivo

doi: 10.1128/Spectrum.00497-21

Figure Lengend Snippet: FOP reduces disease symptoms associated with the burden of Salmonella in OMM 12 mice

Article Snippet: S. enterica subsp. enterica serovar Typhimurium strain ST784 was obtained from Intralytix, Inc. (Columbia, MD), and routinely cultured in lysogeny broth (LB), on LB agar, or on Drigalski agar (Bio-Rad, Hercules, CA) at 37°C.

Techniques:

Journal: Microbiology Spectrum

Article Title: Prophylactic Administration of a Bacteriophage Cocktail Is Safe and Effective in Reducing Salmonella enterica Serovar Typhimurium Burden in Vivo

doi: 10.1128/Spectrum.00497-21

Figure Lengend Snippet: Salmonella infection disturbs the microbiota in OMM 12 mice but FOP does not. (A) Shown is a principal-component analysis (PCA) plot of 16S rRNA gene data obtained from fecal pellets taken at days 0 (baseline), 2 (just before the ST784 challenge), and 6 (before sacrifice) from OMM 12 mice that received FOP ( n = 10) or PBS ( n = 7). All animals were infected with strain ST784 on day 2 (see Fig. 2A for the experimental design). (B) The relative abundances of the ST784 challenge strain and OMM 12 mouse origin bacteria from the fecal samples are shown. A comparison of log 2 -fold changes of 16S rRNA gene read abundances between days and conditions with statistical values is given in Table S3. Note that only 10 of the 12 mouse origin bacteria are detectable by either 16S rRNA gene or quantitative PCR (qPCR), as reported previously .

Article Snippet: S. enterica subsp. enterica serovar Typhimurium strain ST784 was obtained from Intralytix, Inc. (Columbia, MD), and routinely cultured in lysogeny broth (LB), on LB agar, or on Drigalski agar (Bio-Rad, Hercules, CA) at 37°C.

Techniques: Infection, Bacteria, Comparison, Real-time Polymerase Chain Reaction

Journal: Microbiology Spectrum

Article Title: Prophylactic Administration of a Bacteriophage Cocktail Is Safe and Effective in Reducing Salmonella enterica Serovar Typhimurium Burden in Vivo

doi: 10.1128/Spectrum.00497-21

Figure Lengend Snippet: Dose dependence of effect of FOP on ST784 burden in OMM 12 mice. (A) Experimental design. OMM 12 mice ( n = 4 to 6) were orally administered the indicated doses of FOP (10 7 PFU, white squares; 10 8 PFU, gray squares; 10 9 PFU, black squares) on the indicated days. On day 2, all mice were challenged with strain ST784 (1 × 10 8 CFU). (B) Mice were weighed daily. Shown are the percentages of weight loss compared to starting weights of OMM 12 mice over time. (C and D) The amounts of ST784 (CFU) (C) and phages (PFU) (D) in OMM 12 mouse feces were quantitated daily (day 2 corresponds to 3 h after ST784 challenge). (E and F) Intestinal organs were collected on day 6, and the amounts of ST784 (CFU) (E) and phages (PFU) quantitated. Statistical analyses are described in Materials and Methods and reported in Table S4.

Article Snippet: S. enterica subsp. enterica serovar Typhimurium strain ST784 was obtained from Intralytix, Inc. (Columbia, MD), and routinely cultured in lysogeny broth (LB), on LB agar, or on Drigalski agar (Bio-Rad, Hercules, CA) at 37°C.

Techniques:

Journal: PLoS ONE

Article Title: Response of Medicago truncatula Seedlings to Colonization by Salmonella enterica and Escherichia coli O157:H7

doi: 10.1371/journal.pone.0087970

Figure Lengend Snippet: Confocal laser-scanning microscopic image of Medicago roots that were colonized with cocktails of S. enterica (A) or E. coli O157:H7 (B) containing GFP as the reporter. The plants were inoculated with 1×10 4 CFU/plant, and 10 days post-inoculation the roots were fixed with 4% paraformaldehyde and observed under the microscope. Scale bars represent 100 µm. (C) The growth of S. enterica and E. coli O157:H7 cocktails in Luria-Bertani (LB) and Fåhraeus medium at 25°C for 72 hours. The normal growth of these strains was observed in LB medium, whereas no growth was detected in Fåhraeus medium. (D) The growth of cocktails of S. enterica and E. coli O157:H7 in 2-week-old root exudates (RE) of Medicago. Both of these cocktails grew in the 2-week-old root exudates.

Article Snippet: These results indicate that both of the enteric pathogens S. enterica and E. coli O157:H7 are able to enter through the epidermal layer of Medicago roots and colonize the outer cortex very efficiently , .

Techniques: Microscopy

Journal: PLoS ONE

Article Title: Response of Medicago truncatula Seedlings to Colonization by Salmonella enterica and Escherichia coli O157:H7

doi: 10.1371/journal.pone.0087970

Figure Lengend Snippet: Bacterial CFUs recovered from Medicago roots 10 days post-inoculation with cocktails of S. enterica (A) and E. coli O157:H7 (B). The mean value of 5 replicates is represented; error bars represent standard error of the mean. The CFU values were log 10 transformed. (Note: 2 ml of inoculum was used.) (C–E) Longitudinal sections of Medicago roots inoculated with enteric pathogens at 1×10 4 cfu/plant. At 10 days post-inoculation, the roots were fixed with 4% paraformaldehyde, and sections were obtained using a Vibratome®. (C and E) Longitudinal sections of roots inoculated with the S. enterica cocktail. (C) Ep-Epidermis, OC-outer cortex, IC- inner cortex and Va-Vasculature. (D) Longitudinal section of roots inoculated with the E. coli O157:H7 cocktail. Note that both strains ( E. coli O157:H7 expressing either Ds RED or GFP as the visible marker) colonize the same region. Scale bar represents 50 µm for C and 10 µm for D and 20 µm for E.

Article Snippet: These results indicate that both of the enteric pathogens S. enterica and E. coli O157:H7 are able to enter through the epidermal layer of Medicago roots and colonize the outer cortex very efficiently , .

Techniques: Transformation Assay, Expressing, Marker

Journal: PLoS ONE

Article Title: Response of Medicago truncatula Seedlings to Colonization by Salmonella enterica and Escherichia coli O157:H7

doi: 10.1371/journal.pone.0087970

Figure Lengend Snippet: An expression analysis was performed using the Medicago Affymetrix GeneChip® Medicago genome array on 10-day post-inoculated roots inoculated with 2×10 0 CFU/plant (A). The analysis of the microarray data using EB array statistics identified 187 and 168 probe sets that were up-regulated log 2-fold and 58 and 41 probe sets that were down-regulated when treated with S. enterica and E. coli O157, respectively. A total of 71 and 12 probe sets were commonly up-and down-regulated, respectively, in both treatments. An expression analysis of 10-day post-inoculated Medicago plants inoculated with 2×10 0 CFU/plant of a cocktail of either S. enterica (B) or E. coli O157:H7 (C). The relative expression of kinase Medtr2g036460.1 in the plants inoculated with S. enterica (B) and E. coli O157:H7 (C). Error bars represent the standard error of the mean from three biological replicates, and * indicates the significance at the 5% level with a p value of 0.03 and 0.04 for S. enterica and E. coli O157:H7 infection, respectively.

Article Snippet: These results indicate that both of the enteric pathogens S. enterica and E. coli O157:H7 are able to enter through the epidermal layer of Medicago roots and colonize the outer cortex very efficiently , .

Techniques: Expressing, Microarray, Infection

Journal: PLoS ONE

Article Title: Response of Medicago truncatula Seedlings to Colonization by Salmonella enterica and Escherichia coli O157:H7

doi: 10.1371/journal.pone.0087970

Figure Lengend Snippet: Most of the differentially-expressed probe sets involved in response to inoculation with cocktails of S. enterica and E. coli O157:H7 play a role in signaling, secondary metabolites, hormone signaling, cell wall modifications, and pathogenesis-related (PR) proteins (A). Gene enrichment analysis using the AgriGO software platform indicates the functional categories over- and under-represented against the reference (B). (Note: the data for the enrichment analysis were normalized against the reference).

Article Snippet: These results indicate that both of the enteric pathogens S. enterica and E. coli O157:H7 are able to enter through the epidermal layer of Medicago roots and colonize the outer cortex very efficiently , .

Techniques: Software, Functional Assay

Journal: PLoS ONE

Article Title: Response of Medicago truncatula Seedlings to Colonization by Salmonella enterica and Escherichia coli O157:H7

doi: 10.1371/journal.pone.0087970

Figure Lengend Snippet: List of the probe sets related to the biotic stress response that were differentially expressed in both E. coli O157:H7- and S. enterica -inoculated plants.

Article Snippet: These results indicate that both of the enteric pathogens S. enterica and E. coli O157:H7 are able to enter through the epidermal layer of Medicago roots and colonize the outer cortex very efficiently , .

Techniques:

Journal: PLoS ONE

Article Title: Response of Medicago truncatula Seedlings to Colonization by Salmonella enterica and Escherichia coli O157:H7

doi: 10.1371/journal.pone.0087970

Figure Lengend Snippet: List of the probe sets related to the biotic stress response that were differentially expressed in both E. coli O157:H7- and S. enterica -inoculated plants.

Article Snippet: These results indicate that both of the enteric pathogens S. enterica and E. coli O157:H7 are able to enter through the epidermal layer of Medicago roots and colonize the outer cortex very efficiently , .

Techniques:

Journal: Applied and Environmental Microbiology

Article Title: Temperature-Sensitive Salmonella enterica Serovar Enteritidis PT13a Expressing Essential Proteins of Psychrophilic Bacteria

doi: 10.1128/AEM.01953-15

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: S. enterica subsp. enterica serovar Typhimurium DT104 , Wild-type S . Typhimurium , Zoetis strain collection.

Techniques: Plasmid Preparation, Clone Assay, Variant Assay, Transformation Assay

Journal: BMC Microbiology

Article Title: Molecular beacon-based real-time PCR detection of primary isolates of Salmonella Typhimurium and Salmonella Enteritidis in environmental and clinical samples

doi: 10.1186/1471-2180-9-97

Figure Lengend Snippet: Salmonella enterica serovars a

Article Snippet: The reference S. enterica serovars listed in Table were obtained from the Community Reference Laboratory for Salmonella at the National Institute for Public Health and the Environment (RIVM, Bilthoven, the Netherlands).

Techniques:

Journal: BMC Microbiology

Article Title: Molecular beacon-based real-time PCR detection of primary isolates of Salmonella Typhimurium and Salmonella Enteritidis in environmental and clinical samples

doi: 10.1186/1471-2180-9-97

Figure Lengend Snippet: Schematic real-time PCR results for the second step reaction . Representative real-time PCR results as established by the second step multiplex reaction (described in Materials and Methods). The plots show average normalised linear amplification of representative samples shown for demonstration of typical results obtained from S . Typhimurium, S . Enteritidis and other Salmonella samples. With DNA from S . Typhimurium strains, only fliC -specific, HEX-labelled molecular beacons hybridise to the amplicons, generating pink fluorescence, whereas the prot6E -specific, TET-labelled molecular beacons retain their stem-and-loop structure and cannot produce an orange fluorescent signal. With DNA from S . Enteritidis strains, the prot6E -specific, TET-labelled molecular beacons hybridise to their target amplicons and produce an orange fluorescent signal, whereas the fliC -specific, HEX-labelled molecular beacons remain dark. With DNA from other Salmonella serotypes, no target amplicons are detected and both molecular beacons remain dark. The dashed line on the plots represents the normalised threshold for detection of fluorescence, the baseline above which fluorescence increases significantly on amplification and detection of the target sequence.

Article Snippet: The reference S. enterica serovars listed in Table were obtained from the Community Reference Laboratory for Salmonella at the National Institute for Public Health and the Environment (RIVM, Bilthoven, the Netherlands).

Techniques: Real-time Polymerase Chain Reaction, Multiplex Assay, Amplification, Fluorescence, Sequencing

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Active Quantum Biomaterials‐Enhanced Microrobots for Food Safety

doi: 10.1002/smll.202404248

Figure Lengend Snippet: Self‐propelled graphene‐based QBEMRs as active biocarriers for on‐the‐fly detection of S. enterica endotoxin. The scheme shows the QBEMRs with the attached Rhodamine‐labeled affinity peptide before ( OFF in solution) and after ( ON in solution) navigation in an S. enterica endotoxin‐contaminated solution. This initiated the release and interaction of the affinity peptide with the S. enterica endotoxin, resulting in fluorescence recovery in the solution ( ON stage). (For relative charge symbols, see Figure ).

Article Snippet: Citric acid (cat. 251275), nickel tetrahydrate (cat. 262277), nickel chloride hexahydrate (cat. N6136), boric acid (cat. 15665), chloroplatinic acid hydrate (cat. 520896), SDS (cat. 71727), PEG (cat. 89510), Tween 20 (cat. P9416), hydrogen peroxide (30%, cat. 216763) (H 2 O 2 ), Triton X‐100, and endotoxins from S. enterica Typhimurium (cat. L6143) and S. enterica Enteritidis (cat. L7770) were purchased from Merck (Madrid, Spain).

Techniques: Labeling, Fluorescence

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Active Quantum Biomaterials‐Enhanced Microrobots for Food Safety

doi: 10.1002/smll.202404248

Figure Lengend Snippet: A) Z‐potential measurements of GQDs and peptide@QBEMRs biocarriers before and after S. enterica endotoxin interactions: a) QBEMRs; b) RhO‐labeled affinity peptide; c) peptide@QBEMRs; d) S. typhimurium endotoxins; e) peptide@QBEMRs interacting with S. typhimurium endotoxins. B) Histogram illustrating fluorescence intensity controls for (a) RhO‐labeled affinity peptide, b) peptide@QBEMRs, c) peptide@ QBEMRs interacting with S. typhimurium endotoxins, d) S. typhimurium endotoxins. Experimental conditions: λ ex = 560 nm; [peptide, S. enterica endotoxin] = 50 µg mL −1 ; incubation time: 10 min; recovery time: 5 min; 1% PEG; 9% v/v in H 2 O 2 . Error bars represent the standard deviation ( n = 3).

Article Snippet: Citric acid (cat. 251275), nickel tetrahydrate (cat. 262277), nickel chloride hexahydrate (cat. N6136), boric acid (cat. 15665), chloroplatinic acid hydrate (cat. 520896), SDS (cat. 71727), PEG (cat. 89510), Tween 20 (cat. P9416), hydrogen peroxide (30%, cat. 216763) (H 2 O 2 ), Triton X‐100, and endotoxins from S. enterica Typhimurium (cat. L6143) and S. enterica Enteritidis (cat. L7770) were purchased from Merck (Madrid, Spain).

Techniques: Labeling, Fluorescence, Incubation, Standard Deviation

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Active Quantum Biomaterials‐Enhanced Microrobots for Food Safety

doi: 10.1002/smll.202404248

Figure Lengend Snippet: QBEMRs active biocarriers toward the optical on‐the‐fly S. enterica endotoxin determination. A) Calibration plot between the logarithmic (log) values of the concentration of the analyte [endotoxin] versus log [Fluorescence Intensity] under motion, with its corresponding error bars ( n = 3 ). B) Histogram illustrating the recovery in the fluorescence intensities achieved by the peptide@QBEMRs toward a fixed concentration of S. enterica endotoxins (50 µg mL −1 ) under motion (9% H 2 O 2 ) or static (no fuel) conditions. C) Histogram depicting the recovery of fluorescence intensities achieved by the peptide@QBEMRs biocarriers toward different targets (i.e. , E. coli , S. enterica , and S. typhimurium : (50 µg mL −1 ) (9% H 2 O 2 ). D) Histogram showing the recovery obtained through the peptide@QBEMRs in various real samples (orange juice, milk) fortified with 15 µg mL −1 endotoxin ( n = 3 ). Experimental conditions: λ ex = 560 nm; incubation time: 10 min; recovery time: 5 min; 1% PEG; 9% v/v in H 2 O 2 .

Article Snippet: Citric acid (cat. 251275), nickel tetrahydrate (cat. 262277), nickel chloride hexahydrate (cat. N6136), boric acid (cat. 15665), chloroplatinic acid hydrate (cat. 520896), SDS (cat. 71727), PEG (cat. 89510), Tween 20 (cat. P9416), hydrogen peroxide (30%, cat. 216763) (H 2 O 2 ), Triton X‐100, and endotoxins from S. enterica Typhimurium (cat. L6143) and S. enterica Enteritidis (cat. L7770) were purchased from Merck (Madrid, Spain).

Techniques: Concentration Assay, Fluorescence, Incubation

Journal: PLoS Neglected Tropical Diseases

Article Title: Vaccination against the digestive enzyme Cathepsin B using a YS1646 Salmonella enterica Typhimurium vector provides almost complete protection against Schistosoma mansoni challenge in a mouse model

doi: 10.1371/journal.pntd.0007490

Figure Lengend Snippet: Baseline serum was collected on day 0 for all mice. Each group consists of either a saline control, EV→IM, PO→PO, IM→IM, PO→IM, and IM→PO for the nirB_SspH1 and/or the SspH1_SspH1 construct. Mice receive 3 oral doses of YS1646 (1x10 9 cfu/dose) or PBS every other day while others receive an intramuscular dose of 20 μg of CatB on day 5. Mice were bled and underwent a second round of vaccination three weeks later before being challenged with 150 S . mansoni cercariae by tail penetration. All animals were sacrificed 6–7 weeks post-infection.

Article Snippet: S . enterica Typhimurium YS1646 (Cedarlane Labs, Burlington, ON) was cultured in Lysogeny broth (LB) media and strains bearing each construct were generated by electroporation (5ms, 3kV: Biorad, Hercules, CA).

Techniques: Construct, Infection

Journal: PLoS Neglected Tropical Diseases

Article Title: Vaccination against the digestive enzyme Cathepsin B using a YS1646 Salmonella enterica Typhimurium vector provides almost complete protection against Schistosoma mansoni challenge in a mouse model

doi: 10.1371/journal.pntd.0007490

Figure Lengend Snippet: A) The plasmids nirB_SspH1, SspH1_SspH1 and SteA_SteA were transformed into Salmonella strain YS1646. Whole bacteria lysates and monomicrobial culture supernatants were examined for the presence of CatB by western blot. B) The mouse macrophage cell line RAW 264.7 cells were infected with transformed YS1646 strains expressing eGFP as a marker for the capacity of promoter-TSSS pairs to support expression of a foreign protein. DAPI nuclear stain is represented in blue and eGFP is shown in green. Scale at 100 μm. C) Mouse macrophage cells line RAW 264.7 cells were infected with selected plasmids from and the presence of CatB protein was determined by western blotting.

Article Snippet: S . enterica Typhimurium YS1646 (Cedarlane Labs, Burlington, ON) was cultured in Lysogeny broth (LB) media and strains bearing each construct were generated by electroporation (5ms, 3kV: Biorad, Hercules, CA).

Techniques: Transformation Assay, Western Blot, Infection, Expressing, Marker, Staining