Structured Review

Difco s typhimurium
MET-1 pre-treatment does not prevent S. <t>typhimurium</t> intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.
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1) Product Images from "Administration of defined microbiota is protective in a murine Salmonella infection model"

Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

Journal: Scientific Reports

doi: 10.1038/srep16094

MET-1 pre-treatment does not prevent S. typhimurium intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.
Figure Legend Snippet: MET-1 pre-treatment does not prevent S. typhimurium intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.

Techniques Used: Incubation, Infection

MET-1 pretreatment prevented the disruption of ZO-1 and claudin-1 cellular localization in the cecum caused by S. typhimurium . ( a ) ZO-1 immunofluorescence staining (green) in ceca fixed in 10% formalin (400× magnification). Nuclei were stained using Hoechst (blue). White arrows indicate green staining of ZO-1 protein at the membrane of intestinal epithelial cells. VC = uninfected mice pretreated with vehicle control; MET-1 = uninfected mice pretreated with MET-1; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1. VC + S . Tm. mice had a loss of ZO-1 membrane staining that was attenuated in MET-1 + S . Tm. mice. ( b ) Expression of ZO-1 measured by Western blot analysis (S, Triton X-soluble fraction; I, Triton X-insoluble fraction, sample blot shown in top right of panel). Consistent with data in panel ( a ), densitometric values calculated for the ratio of ZO-1 to β-actin showed that MET-1 + S . Tm. mice had increased expression of ZO-1 in insoluble fractions compared to VC + S . Tm. mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction (*p
Figure Legend Snippet: MET-1 pretreatment prevented the disruption of ZO-1 and claudin-1 cellular localization in the cecum caused by S. typhimurium . ( a ) ZO-1 immunofluorescence staining (green) in ceca fixed in 10% formalin (400× magnification). Nuclei were stained using Hoechst (blue). White arrows indicate green staining of ZO-1 protein at the membrane of intestinal epithelial cells. VC = uninfected mice pretreated with vehicle control; MET-1 = uninfected mice pretreated with MET-1; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1. VC + S . Tm. mice had a loss of ZO-1 membrane staining that was attenuated in MET-1 + S . Tm. mice. ( b ) Expression of ZO-1 measured by Western blot analysis (S, Triton X-soluble fraction; I, Triton X-insoluble fraction, sample blot shown in top right of panel). Consistent with data in panel ( a ), densitometric values calculated for the ratio of ZO-1 to β-actin showed that MET-1 + S . Tm. mice had increased expression of ZO-1 in insoluble fractions compared to VC + S . Tm. mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction (*p

Techniques Used: Immunofluorescence, Staining, Mouse Assay, Infection, Expressing, Western Blot

MET-1 inhibited S. typhimurium in vitro but did not inhibit its growth in vivo . ( a ) MET-1 (1:2, 1:100 and 1:300 dilutions) was incubated with S. typhimurium ( S . Tm.), aliquots were withdrawn at times indicated, plated, counted, and plotted as log 10 CFU. MET-1 inhibited the growth of S. Tm. relative to the S . Tm. control, although MET-1 was unable to completely kill S. Tm. even at a high (1:2) concentration. ( b ) Colons were harvested 48 hours after S. Tm. infection, homogenized in sterile PBS, and plated. Infected mice pretreated with MET-1 (MET-1 + S . Tm.) showed no significant reduction of S. Tm. counts in the colon (p > 0.05) compared to mice pretreated with vehicle control (VC + S . Tm.). S. Tm. was not detected in the colon of uninfected mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction. VC = uninfected mice pretreated with vehicle control, n = 18; MET-1 = uninfected mice pretreated with MET-1, n = 18; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 22; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 21.
Figure Legend Snippet: MET-1 inhibited S. typhimurium in vitro but did not inhibit its growth in vivo . ( a ) MET-1 (1:2, 1:100 and 1:300 dilutions) was incubated with S. typhimurium ( S . Tm.), aliquots were withdrawn at times indicated, plated, counted, and plotted as log 10 CFU. MET-1 inhibited the growth of S. Tm. relative to the S . Tm. control, although MET-1 was unable to completely kill S. Tm. even at a high (1:2) concentration. ( b ) Colons were harvested 48 hours after S. Tm. infection, homogenized in sterile PBS, and plated. Infected mice pretreated with MET-1 (MET-1 + S . Tm.) showed no significant reduction of S. Tm. counts in the colon (p > 0.05) compared to mice pretreated with vehicle control (VC + S . Tm.). S. Tm. was not detected in the colon of uninfected mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction. VC = uninfected mice pretreated with vehicle control, n = 18; MET-1 = uninfected mice pretreated with MET-1, n = 18; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 22; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 21.

Techniques Used: In Vitro, In Vivo, Incubation, Concentration Assay, Infection, Mouse Assay

Pro-inflammatory cytokine levels were reduced in S. typhimurium -infected mice pretreated with MET-1. ( a ) Serum cytokine levels were measured 48 hours post-infection using a Bio-Plex Pro mouse cytokine magnetic bead kit. Of the cytokines measured, TNF-α, IL-1β, IL-12p40, IL-12p70, GM-CSF, eotaxin, MIP-1β, and MCP-1 were all significantly reduced in infected mice pretreated with MET-1 (MET-1 + S . Tm.) compared to infected mice pretreated with vehicle control (VC + S . Tm.). There were no significant differences in cytokine concentrations between uninfected mice pretreated with MET-1 or vehicle control (data not shown). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 7 for each group (*p
Figure Legend Snippet: Pro-inflammatory cytokine levels were reduced in S. typhimurium -infected mice pretreated with MET-1. ( a ) Serum cytokine levels were measured 48 hours post-infection using a Bio-Plex Pro mouse cytokine magnetic bead kit. Of the cytokines measured, TNF-α, IL-1β, IL-12p40, IL-12p70, GM-CSF, eotaxin, MIP-1β, and MCP-1 were all significantly reduced in infected mice pretreated with MET-1 (MET-1 + S . Tm.) compared to infected mice pretreated with vehicle control (VC + S . Tm.). There were no significant differences in cytokine concentrations between uninfected mice pretreated with MET-1 or vehicle control (data not shown). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 7 for each group (*p

Techniques Used: Infection, Mouse Assay

Translocation of S. typhimurium to the spleen was reduced in MET-1 pretreated mice. Spleens were harvested 48 hours after S. typhimurium infection, weighed, and homogenized in 1 mL PBS. Serial dilutions were plated on MacConkey agar plates containing 100 μg/mL streptomycin. Plates were incubated at 37 °C for 24 hrs, and the resulting colonies were counted. Infected mice pretreated with MET-1 (MET-1 + S . Tm., n = 13) had reduced S. typhimurium counts in the spleen (*p
Figure Legend Snippet: Translocation of S. typhimurium to the spleen was reduced in MET-1 pretreated mice. Spleens were harvested 48 hours after S. typhimurium infection, weighed, and homogenized in 1 mL PBS. Serial dilutions were plated on MacConkey agar plates containing 100 μg/mL streptomycin. Plates were incubated at 37 °C for 24 hrs, and the resulting colonies were counted. Infected mice pretreated with MET-1 (MET-1 + S . Tm., n = 13) had reduced S. typhimurium counts in the spleen (*p

Techniques Used: Translocation Assay, Mouse Assay, Infection, Incubation

MET-1 attenuates local neutrophil infiltration induced by S. typhimurium infection in cecum (a) Representative images of myeloperoxidase (MPO) immunofluorescence staining for neutrophils (green) in ceca. The area in the box (left) is displayed at a higher magnification in the panels on the right. Nucleic acid was stained using Hoechst (blue). S. typhimurium -infected mice pretreated with vehicle control (VC + S . Tm.) had more abundant immunostaining in the ceca compared to uninfected controls (VC), and was significantly reduced in the ceca of infected mice pretreated with MET-1 (MET-1 + S . Tm. mice). No MPO staining was noted in secondary antibody control slides for each group (not shown). ( b ) Quantification of MPO immunostaining, shown by the ratio of MPO (green immunofluorescence) staining to number of cells (blue nuclei). Data analyzed using 1-way ANOVA with Bonferroni correction, ***p
Figure Legend Snippet: MET-1 attenuates local neutrophil infiltration induced by S. typhimurium infection in cecum (a) Representative images of myeloperoxidase (MPO) immunofluorescence staining for neutrophils (green) in ceca. The area in the box (left) is displayed at a higher magnification in the panels on the right. Nucleic acid was stained using Hoechst (blue). S. typhimurium -infected mice pretreated with vehicle control (VC + S . Tm.) had more abundant immunostaining in the ceca compared to uninfected controls (VC), and was significantly reduced in the ceca of infected mice pretreated with MET-1 (MET-1 + S . Tm. mice). No MPO staining was noted in secondary antibody control slides for each group (not shown). ( b ) Quantification of MPO immunostaining, shown by the ratio of MPO (green immunofluorescence) staining to number of cells (blue nuclei). Data analyzed using 1-way ANOVA with Bonferroni correction, ***p

Techniques Used: Infection, Immunofluorescence, Staining, Mouse Assay, Immunostaining

MET-1 attenuated systemic markers of disease in S. typhimurium -infected mice. ( a ) MET-1 attenuated weight loss in S. typhimurium infected mice. Following oral infection with S. typhimurium , mice were weighed daily and the percent change in weight from 0 to 48 hours is shown. VC = uninfected mice pretreated with vehicle control, n = 16; MET-1 = uninfected mice pretreated with MET-1, n = 16; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 18; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 18. Data were analyzed with a 2-way ANOVA using Bonferroni correction (*p
Figure Legend Snippet: MET-1 attenuated systemic markers of disease in S. typhimurium -infected mice. ( a ) MET-1 attenuated weight loss in S. typhimurium infected mice. Following oral infection with S. typhimurium , mice were weighed daily and the percent change in weight from 0 to 48 hours is shown. VC = uninfected mice pretreated with vehicle control, n = 16; MET-1 = uninfected mice pretreated with MET-1, n = 16; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 18; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 18. Data were analyzed with a 2-way ANOVA using Bonferroni correction (*p

Techniques Used: Infection, Mouse Assay

MET-1 did not enhance intestinal mucin production in ceca of S. typhimurium - infected mice. ( a ) Alcian blue staining (blue) of ceca fixed in 10% formalin were counterstained with nuclear red fast solution (red). ( b ) Goblet cells were enumerated from 10 random high-powered field views (400× magnification) spanning from muscularis mucosa to surface epithelium on Alcian blue stained sections. Infected mice pretreated with vehicle control (VC + S . Tm.) had less mucin staining than uninfected mice pretreated with vehicle control (VC + S . Tm.). However, there was no statistically significant difference between infected mice pretreated with MET-1 (MET-1 + S . Tm.) and vehicle control (VC + S . Tm.). ( c ) MUC2 immunofluorescence staining (green) in ceca fixed in Methanol-Carnoy’s solution. Nucleic acid was stained using DAPI (blue). Uninfected mice pretreated with VC or MET-1 had similar localization of MUC2 staining that differed from VC + S . Tm. mice. Again, MUC2 staining in MET-1 + S . Tm. mice more closely resembled VC + S . Tm. mice than uninfected controls. Quantification shown in (d). Data were analyzed using a 1-way ANOVA with Bonferroni correction. n = 3, (**p
Figure Legend Snippet: MET-1 did not enhance intestinal mucin production in ceca of S. typhimurium - infected mice. ( a ) Alcian blue staining (blue) of ceca fixed in 10% formalin were counterstained with nuclear red fast solution (red). ( b ) Goblet cells were enumerated from 10 random high-powered field views (400× magnification) spanning from muscularis mucosa to surface epithelium on Alcian blue stained sections. Infected mice pretreated with vehicle control (VC + S . Tm.) had less mucin staining than uninfected mice pretreated with vehicle control (VC + S . Tm.). However, there was no statistically significant difference between infected mice pretreated with MET-1 (MET-1 + S . Tm.) and vehicle control (VC + S . Tm.). ( c ) MUC2 immunofluorescence staining (green) in ceca fixed in Methanol-Carnoy’s solution. Nucleic acid was stained using DAPI (blue). Uninfected mice pretreated with VC or MET-1 had similar localization of MUC2 staining that differed from VC + S . Tm. mice. Again, MUC2 staining in MET-1 + S . Tm. mice more closely resembled VC + S . Tm. mice than uninfected controls. Quantification shown in (d). Data were analyzed using a 1-way ANOVA with Bonferroni correction. n = 3, (**p

Techniques Used: Infection, Mouse Assay, Staining, Immunofluorescence

2) Product Images from "Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice"

Article Title: Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0074963

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display an increased cecal macrophage infiltration. (A) Formalin fixed paraffin embedded cecum tissue was thin sectioned at 2 µm. Macrophages were stained by targeting the F4/80 receptor expressed on mouse macrophages using immunohistochemistry with specific antibodies. Brown color indicates positively stained macrophages. Magnification 400-fold. Bar = 100 µm. (B) Positively stained macrophages were enumerated along a stretch of 50 µm of lamina muscularis for both lamina propria and sub-mucosa (see materials and methods). SIHUMI mice colonized with both A. muciniphila and S. Typhimurium had the highest macrophage infiltration scores compared to the other groups (see Figure. 1). Data are expressed as median with range. n = 5 mice per group. *P

Techniques Used: Mouse Assay, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry

Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P
Figure Legend Snippet: Concomitant presence of A. muciniphila and S. Typhimurium results in increased histopathology scores in SIHUMI mice. (A) Gnotobiotic C3H mice containing 8 defined microbial species (SIHUMI) were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). SIHUMI and SIHUMI-A mice had the lowest histopathology scores (≤4.0) with no signs of inflammation and were therefore taken as baseline (dotted line). Data are expressed as median with range. *P

Techniques Used: Histopathology, Mouse Assay

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display enlarged mLN and elevated S. Typhimurium cell numbers. (A) Mesenteric lymph nodes (mLN) were obtained from four groups of gnotobiotic C3H mice. SIHUMI mice were subsequently inoculated with A. muciniphila or S. Typhimurium or consecutively with both organisms (see Figure 1 ). The mLN tissue was homogenized and DNA was isolated to quantify S. Typhimurium using quantitative PCR with primers targeting the ttr-region of S. Typhimurium. Absolute cell numbers were calculated based on calibration curves with known concentrations of S. Typhimurium. The mLN of SIHUMI-AS mice contained 10-fold higher cell numbers of S. Typhimurium compared to SIHUMI-S mice. Data are expressed as mean±standard error. n = 10 mice per group. *P

Techniques Used: Mouse Assay, Isolation, Real-time Polymerase Chain Reaction

SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.
Figure Legend Snippet: SIHUMI mice colonized with both A. muciniphila and S. Typhimurium display reduced mucus sulphation. Formalin fixed thin sections (4 µm) of cecal tissue of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1) were stained with high iron diamine (HID)/AB at pH-2.5 and subsequently analyzed. Brown color indicates sulphated mucins while blue color indicates sialylated mucins. SIHUMI-AS mice display few sulphated mucins compared to the other mouse groups. Magnification 400×. Bars indicate 100 µm.

Techniques Used: Mouse Assay, Staining

Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).
Figure Legend Snippet: Hypothetical Scheme. The presence of A. muciniphila, leads to the exacerbation of S. Typhimurium-induced intestinal inflammation. We propose that the presence of A. muciniphila causes changes in mucin composition and production, which in turn facilitates the invasion of S. Typhimurium into the host. Increased inflammatory status was characterized by increased pro-inflammatory cytokines, increased macrophage infiltration and invasion of the pathogen into the lymph nodes, reduced number of mucin-filled goblet cells in SIHUMI-AS mice (B) compared to SIHUMI-S mice (A). Our data suggests that in the presence of both A. muciniphila and S. Typhimurium, mucus sulphation is diminished and this may facilitate the access of S. Typhimurium to sialic acid in mucus. Sialic acid may serve as a substrate and adhesion site for S. Typhimurium in the gut [56] , [57] . Increased gene expression of IFN-γ and IP-10 indicate an increased NK-cell recruitment. mLN - mesenteric lymph nodes, NK- Natural killer cells. (↑ increased; ↓ decreased; grey dotted line: assumed processes including lectin-sialic acid binding [56] , M-cells for pathogen transit [43] , [58] , [59] ; black line: supported by data of the present study).

Techniques Used: Mouse Assay, Expressing, Binding Assay

Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .
Figure Legend Snippet: Presence of A. muciniphila renders S. Typhimurium the dominant species in gnotobiotic SIHUMI mice. Cecal contents were collected from gnotobiotic C3H mice, differing in their microbial status: (A) Mice with a defined microbial community of eight bacterial species (SIHUMI), (B) SIHUMI mice additionally colonized with A. muciniphila (SIHUMI-A), (C) SIHUMI mice infected with S. Typhimurium (SIHUMI-S) and (D) SIHUMI mice colonized with A. muciniphila and 10 days later infected with S. Typhimurium (SIHUMI-AS) (see Figure 1 ). Total DNA was extracted and bacterial cell numbers were quantified by qPCR with primers targeting the HSP60 gene of the SIHUMI members, the 16S rRNA gene of A. muciniphila and the ttr-region of S. Typhimurium. Calculation of the cell numbers was based on DNA obtained from cell suspensions containing known cell numbers of the targeted bacterial species (see materials and methods). Presence of A. muciniphila in SIHUMI-AS mice is attributed to an increase in the proportion of S. Typhimurium cells at the expense of other community members showing reduced proportion of SIHUMI members. Ten animals per group were used. The bacterial cell numbers and P-values for the differences between the groups are provided in Table 1 .

Techniques Used: Mouse Assay, Infection, Real-time Polymerase Chain Reaction

Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P
Figure Legend Snippet: Presence of both A. muciniphila and S. Typhimurium is accompanied by increased pro-inflammatory cytokines. (A) Cecal mRNA levels of IFN-γ, IP-10, TNF-α, IL-12, IL-6, IL-17 and IL-18 in gnotobiotic SIHUMI mice were measured. mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS (see Figure. 1). The mRNA was converted to cDNA for quantitative real-time PCR measurement (see materials and methods). Inoculation of the gnotobiotic SIHUMI mice with A. muciniphila followed by S. Typhimurium infection (SIHUMI-AS) caused an increase in mRNA levels of pro-inflammatory cytokines except IL-18. Data are expressed as mean±standard error. n = 6 per group. Star indicates statistically significant differences ( *P

Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Infection

SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P
Figure Legend Snippet: SIHUMI mice with both A. muciniphila and S. Typhimurium display increased MUC2 mRNA levels (A) and reduced numbers of mucin filled goblet cells (B and C). (A) mRNA was extracted from cecum mucosa of mice belonging to either one of four groups: SIHUMI, SIHUMI-A, SIHUMI-S and SIHUMI-AS. MUC2 mRNA from cecum mucosa was converted to cDNA and expression levels were quantified using real-time PCR (see materials and methods). SIHUMI-A and SIHUMI-AS mice showed significantly higher MUC2 gene expression compared to the other two groups, harboring no A. muciniphila . Data are expressed as mean±standard error. n = 6 per group. *P

Techniques Used: Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.
Figure Legend Snippet: Design of the animal experiment. Fourty C3H mice associated with a defined microbial community of 8 bacterial species (SIHUMI) were allocated to four different groups (10 mice per group). Each mouse was associated with 8 bacterial species (SIHUMI). Twelve weeks-old SIHUMI mice were subsequently associated with A. muciniphila (SIHUMI-A) or S. Typhimurium (SIHUMI-S) or with both A. muciniphila and S. Typhimurium (SIHUMI-AS). SIHUMI mice received only sterile medium. Times of association, infection and killing are as indicated. ‡ - killed.

Techniques Used: Mouse Assay, Infection

3) Product Images from "Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium"

Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

Journal: mBio

doi: 10.1128/mBio.01040-18

Virulence of Δ zntA Δ zitB mutant S. Typhimurium is attenuated in NO·-producing mice. Solid lines represent the median competitive index (CI) for each organ. The dotted line represents the expected CI if neither strain has a competitive advantage. (A) In wild-type C3H/HeOuJ mice, a Δ zntA Δ zitB mutant has a significant competitive disadvantage compared to wild type ( P = 0.002 by Wilcoxon signed-rank test to a hypothetical median of 1 for both spleen and liver). (B) In C3H/HeOuJ mice that cannot produce NO· due to treatment with 500 µg ml −1 l - N 6 -(1-iminoethyl)lysine dihydrochloride ( l -NIL), the mutant no longer has a statistically significant disadvantage compared to wild type, and the CIs are significantly different (*, P = 0.007 for spleen and P
Figure Legend Snippet: Virulence of Δ zntA Δ zitB mutant S. Typhimurium is attenuated in NO·-producing mice. Solid lines represent the median competitive index (CI) for each organ. The dotted line represents the expected CI if neither strain has a competitive advantage. (A) In wild-type C3H/HeOuJ mice, a Δ zntA Δ zitB mutant has a significant competitive disadvantage compared to wild type ( P = 0.002 by Wilcoxon signed-rank test to a hypothetical median of 1 for both spleen and liver). (B) In C3H/HeOuJ mice that cannot produce NO· due to treatment with 500 µg ml −1 l - N 6 -(1-iminoethyl)lysine dihydrochloride ( l -NIL), the mutant no longer has a statistically significant disadvantage compared to wild type, and the CIs are significantly different (*, P = 0.007 for spleen and P

Techniques Used: Mutagenesis, Mouse Assay

Expression of zinc transport systems in S. Typhimurium. qPCR data are presented as a positive fold change of treated compared to untreated cells with zinc efflux systems shown in blue and zinc acquisition systems shown in red. The solid line indicates a fold change of 1 to delineate between upregulation ( > 1) and downregulation (
Figure Legend Snippet: Expression of zinc transport systems in S. Typhimurium. qPCR data are presented as a positive fold change of treated compared to untreated cells with zinc efflux systems shown in blue and zinc acquisition systems shown in red. The solid line indicates a fold change of 1 to delineate between upregulation ( > 1) and downregulation (

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

ICP-MS analysis of total cellular zinc content in S. Typhimurium following NO· treatment and transcriptional monitoring of NO· sensed by cells. (A) S. Typhimurium cells at an OD 600 of ≈1 were treated with 2 mM diethylamine NONOate (DEANO), and total cellular zinc was measured at various times posttreatment. By 5 min posttreatment, total cellular zinc had fallen significantly compared to untreated cells, suggesting that zinc is effluxed from the cell following NO· treatment. Zinc levels gradually recovered to baseline levels over the course of 60 min. Statistical significance was determined by unpaired two-tailed t test; * indicates P values of
Figure Legend Snippet: ICP-MS analysis of total cellular zinc content in S. Typhimurium following NO· treatment and transcriptional monitoring of NO· sensed by cells. (A) S. Typhimurium cells at an OD 600 of ≈1 were treated with 2 mM diethylamine NONOate (DEANO), and total cellular zinc was measured at various times posttreatment. By 5 min posttreatment, total cellular zinc had fallen significantly compared to untreated cells, suggesting that zinc is effluxed from the cell following NO· treatment. Zinc levels gradually recovered to baseline levels over the course of 60 min. Statistical significance was determined by unpaired two-tailed t test; * indicates P values of

Techniques Used: Mass Spectrometry, Two Tailed Test

Free intracellular zinc levels increase in a Δ zntA Δ zitB S. Typhimurium mutant during macrophage infection in response to NO· production. (A) Changes in NO· production are shown at the time of infection (0 h) and 13 h postinfection in the presence or absence of the NOS inhibitor l -NMMA. IFN-γ-primed murine macrophages infected with either wild-type (blue) or Δ zntA Δ zitB (green) S. Typhimurium produced significant levels of NO· after 13 h in the absence of the NOS inhibitor l -NMMA ( P
Figure Legend Snippet: Free intracellular zinc levels increase in a Δ zntA Δ zitB S. Typhimurium mutant during macrophage infection in response to NO· production. (A) Changes in NO· production are shown at the time of infection (0 h) and 13 h postinfection in the presence or absence of the NOS inhibitor l -NMMA. IFN-γ-primed murine macrophages infected with either wild-type (blue) or Δ zntA Δ zitB (green) S. Typhimurium produced significant levels of NO· after 13 h in the absence of the NOS inhibitor l -NMMA ( P

Techniques Used: Mutagenesis, Infection, Produced

ZntA and ZitB are the primary zinc efflux transporters in S. Typhimurium. (A) A Δ zntA mutant (pink) was impaired for growth, represented by a delayed exit from lag phase, in both 0.125 mM ZnSO 4 and 0.25 mM ZnSO 4 , while a Δ zntB mutant (aqua) and a Δ zitB mutant (light green) exhibited growth comparable to wild type (blue) under these conditions. (B) Significance of the growth defect for the Δ zntA mutant in panel A was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. (C) A Δ zntB Δ zitB double mutant (purple) exhibited growth comparable to wild type (blue) when exposed to elevated zinc concentrations. A Δ zntA Δ zntB mutant (red) behaved similarly to a Δ zntA mutant, whereas a Δ zntA Δ zitB mutant (green) displayed a more severe growth delay at 0.125 mM ZnSO 4 and was unable to grow at 0.25 mM ZnSO 4 . A Δ zntA Δ zntB Δ zitB triple mutant (orange) exhibited growth characteristics identical to those of a Δ zntA Δ zitB double mutant. (D) Significance of the growth defects for the mutants in panel C was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. Statistical significance of the growth defects was determined by unpaired two-tailed t test, and an asterisk indicates P values of
Figure Legend Snippet: ZntA and ZitB are the primary zinc efflux transporters in S. Typhimurium. (A) A Δ zntA mutant (pink) was impaired for growth, represented by a delayed exit from lag phase, in both 0.125 mM ZnSO 4 and 0.25 mM ZnSO 4 , while a Δ zntB mutant (aqua) and a Δ zitB mutant (light green) exhibited growth comparable to wild type (blue) under these conditions. (B) Significance of the growth defect for the Δ zntA mutant in panel A was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. (C) A Δ zntB Δ zitB double mutant (purple) exhibited growth comparable to wild type (blue) when exposed to elevated zinc concentrations. A Δ zntA Δ zntB mutant (red) behaved similarly to a Δ zntA mutant, whereas a Δ zntA Δ zitB mutant (green) displayed a more severe growth delay at 0.125 mM ZnSO 4 and was unable to grow at 0.25 mM ZnSO 4 . A Δ zntA Δ zntB Δ zitB triple mutant (orange) exhibited growth characteristics identical to those of a Δ zntA Δ zitB double mutant. (D) Significance of the growth defects for the mutants in panel C was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. Statistical significance of the growth defects was determined by unpaired two-tailed t test, and an asterisk indicates P values of

Techniques Used: Mutagenesis, Two Tailed Test

A model of zinc homeostasis in Salmonella Typhimurium. Under conditions of zinc deficiency, zinc is not available to bind to the Zur repressor, leading to expression of the ZnuABC zinc acquisition system. ZupT, whose regulation is uncharacterized, has also been shown to contribute to zinc acquisition. When zinc is abundant, Zur bound to zinc represses ZnuABC expression. In addition, free cytoplasmic zinc binds to the transcriptional activator ZntR to induce expression of the ZntA zinc efflux system. Zinc efflux in S. Typhimurium is also mediated by ZitB. Under conditions of nitrosative stress, S -nitrosylation of cysteine ligands in zinc metalloproteins leads to mobilization of free intracellular zinc. The zinc efflux activities of ZntA and ZitB are required for the resistance of S. Typhimurium to nitrosative stress.
Figure Legend Snippet: A model of zinc homeostasis in Salmonella Typhimurium. Under conditions of zinc deficiency, zinc is not available to bind to the Zur repressor, leading to expression of the ZnuABC zinc acquisition system. ZupT, whose regulation is uncharacterized, has also been shown to contribute to zinc acquisition. When zinc is abundant, Zur bound to zinc represses ZnuABC expression. In addition, free cytoplasmic zinc binds to the transcriptional activator ZntR to induce expression of the ZntA zinc efflux system. Zinc efflux in S. Typhimurium is also mediated by ZitB. Under conditions of nitrosative stress, S -nitrosylation of cysteine ligands in zinc metalloproteins leads to mobilization of free intracellular zinc. The zinc efflux activities of ZntA and ZitB are required for the resistance of S. Typhimurium to nitrosative stress.

Techniques Used: Expressing

Zinc efflux by ZntA and ZitB is required for S. Typhimurium resistance to nitrosative stress in vitro . (A) Single zinc efflux mutants (pink, aqua, and light green) and a Δ zntA Δ zntB double mutant (red) were no more sensitive to NO· generated by the donor spermine NONOate (SperNO) than wild-type cells (blue). However, a Δ zntA Δ zitB double mutant (green) exhibited significantly delayed growth, indicating enhanced NO· sensitivity (*, P
Figure Legend Snippet: Zinc efflux by ZntA and ZitB is required for S. Typhimurium resistance to nitrosative stress in vitro . (A) Single zinc efflux mutants (pink, aqua, and light green) and a Δ zntA Δ zntB double mutant (red) were no more sensitive to NO· generated by the donor spermine NONOate (SperNO) than wild-type cells (blue). However, a Δ zntA Δ zitB double mutant (green) exhibited significantly delayed growth, indicating enhanced NO· sensitivity (*, P

Techniques Used: In Vitro, Mutagenesis, Generated

Classification of proteins in the S. Typhimurium S -nitrosoproteome. (A) Functional classification of proteins with cysteine residues modified by NO· treatment. A total of 141 modified proteins were identified, and classification is shown as a percentage of the total. (B) Of the proteins modified by NO·, 15 (~10%) were found to be zinc metalloproteins. The metalloproteins are sorted by functional category and listed in numerical order by gene identifier in S. Typhimurium strain 14028s.
Figure Legend Snippet: Classification of proteins in the S. Typhimurium S -nitrosoproteome. (A) Functional classification of proteins with cysteine residues modified by NO· treatment. A total of 141 modified proteins were identified, and classification is shown as a percentage of the total. (B) Of the proteins modified by NO·, 15 (~10%) were found to be zinc metalloproteins. The metalloproteins are sorted by functional category and listed in numerical order by gene identifier in S. Typhimurium strain 14028s.

Techniques Used: Functional Assay, Modification

4) Product Images from "Outer membrane vesicles from flagellin-deficient Salmonella enterica serovar Typhimurium induce cross-reactive immunity and provide cross-protection against heterologous Salmonella challenge"

Article Title: Outer membrane vesicles from flagellin-deficient Salmonella enterica serovar Typhimurium induce cross-reactive immunity and provide cross-protection against heterologous Salmonella challenge

Journal: Scientific Reports

doi: 10.1038/srep34776

IgG and secretory IgA (S-IgA) immune responses were analysed in sera from mice immunized with OMVs. The total amount of anti-LPS ( a ) or -OMP ( b ) IgG in the sera obtained from mice immunized intranasally with OMVs, the total amount of anti-LPS ( c ) or -OMP ( d ) IgG in the sera obtained from mice immunized intraperitoneally with OMVs, and the total amount of S-IgA that was specific for LPS ( e ) or for OMPs ( f ) were measured using quantitative ELISA. Each group consisted of 10 (control) or 12 (vaccinated) mice. The mice were immunized with OMVs that were derived from S. Typhimurium and then boosted at week 5. Samples were collected at 4 weeks and 8 weeks after the first immunization. PBS-vaccinated mice served as the control group. The data shown represent the concentration of IgG or S-IgA antibodies in samples obtained from mice and are shown according to standard curves.
Figure Legend Snippet: IgG and secretory IgA (S-IgA) immune responses were analysed in sera from mice immunized with OMVs. The total amount of anti-LPS ( a ) or -OMP ( b ) IgG in the sera obtained from mice immunized intranasally with OMVs, the total amount of anti-LPS ( c ) or -OMP ( d ) IgG in the sera obtained from mice immunized intraperitoneally with OMVs, and the total amount of S-IgA that was specific for LPS ( e ) or for OMPs ( f ) were measured using quantitative ELISA. Each group consisted of 10 (control) or 12 (vaccinated) mice. The mice were immunized with OMVs that were derived from S. Typhimurium and then boosted at week 5. Samples were collected at 4 weeks and 8 weeks after the first immunization. PBS-vaccinated mice served as the control group. The data shown represent the concentration of IgG or S-IgA antibodies in samples obtained from mice and are shown according to standard curves.

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Concentration Assay

Internalization of OMVs by RAW264.7 macrophage cells. OMVs (2 μg/ml final concentration) were incubated with cells for 12 hours. OMVs were stained by 1% (vol/vol) lipophilic fluorophore dialkylcarbocyanine iodide (Dil) (red), and the nuclei (blue) were stained by DAPI. The results were recorded using an AMG EVOS digital inverted multi-functional microscope (AMG) at 100x magnification. Arrows indicate the internalization of OMVs derived from S. Typhimurium by macrophages.
Figure Legend Snippet: Internalization of OMVs by RAW264.7 macrophage cells. OMVs (2 μg/ml final concentration) were incubated with cells for 12 hours. OMVs were stained by 1% (vol/vol) lipophilic fluorophore dialkylcarbocyanine iodide (Dil) (red), and the nuclei (blue) were stained by DAPI. The results were recorded using an AMG EVOS digital inverted multi-functional microscope (AMG) at 100x magnification. Arrows indicate the internalization of OMVs derived from S. Typhimurium by macrophages.

Techniques Used: Concentration Assay, Incubation, Staining, Functional Assay, Microscopy, Derivative Assay

Survival in vaccinated mice after oral challenge with wild-type S. Typhimurium. Intranasal ( a ) and intraperitoneal ( b ) immunization with OMVs derived from S. Typhimurium flagellin-deficient mutants provided protection against oral challenge with wild-type S . Typhimurium in BALB/c mice. In total, 10 (control) or 12 (vaccinated) mice per group were immunized twice at 4-week intervals with the indicated OMVs. The mice were challenged with 10 9 CFU of S. Typhimurium at 5 weeks after the boost immunization. Mortality was monitored for 3 weeks after challenge. The numbers in parentheses refer to the number of surviving mice and the total number of mice per group. All vaccine groups were significantly different from the PBS control group ( P
Figure Legend Snippet: Survival in vaccinated mice after oral challenge with wild-type S. Typhimurium. Intranasal ( a ) and intraperitoneal ( b ) immunization with OMVs derived from S. Typhimurium flagellin-deficient mutants provided protection against oral challenge with wild-type S . Typhimurium in BALB/c mice. In total, 10 (control) or 12 (vaccinated) mice per group were immunized twice at 4-week intervals with the indicated OMVs. The mice were challenged with 10 9 CFU of S. Typhimurium at 5 weeks after the boost immunization. Mortality was monitored for 3 weeks after challenge. The numbers in parentheses refer to the number of surviving mice and the total number of mice per group. All vaccine groups were significantly different from the PBS control group ( P

Techniques Used: Mouse Assay, Derivative Assay

Cross-reactivity of OMVs derived from flagellin-deficient S. Typhimurium mutant strain. Cross-reactivity of IgG in sera obtained from intranasally ( a ) or intraperitoneally ( b ) immunized mice against OMPs from other serotypes of Salmonella , including S. Choleraesuis and S. Enteritidis, to analyse OMV-induced cross-protection. Each vaccinated group consisted of 12 mice, and the PBS group consisted of 10 mice. The cross-reactivity data represent the exact concentration of total IgG antibodies in the sera, as quantified using the corresponding standard curve using individual sera obtained from mice immunized intranasally or intraperitoneally with OMVs derived from S. Typhimurium. The error bars represent variations between triplicate wells. Competitive ELISA to determine the cross-reactivity of OMVs derived from flagellin-deficient mutant against heterologous Salmonella. OMPs isolated from S. Typhimurium were incubated into plates as coating antigen. OMPs from S. Choleraesuis, S. Enteritidis or S. Typhimurium (control) as the competitive antigen diluted from 1/10 to 1/ 7,290 were incubated in wells. The sera were obtained from mice (n = 10 or 12) immunized intranasally ( c ) or intraperitoneally ( d ) with OMVs at 8 weeks after the first immunization. The error bars represent variations from triplicate wells. P
Figure Legend Snippet: Cross-reactivity of OMVs derived from flagellin-deficient S. Typhimurium mutant strain. Cross-reactivity of IgG in sera obtained from intranasally ( a ) or intraperitoneally ( b ) immunized mice against OMPs from other serotypes of Salmonella , including S. Choleraesuis and S. Enteritidis, to analyse OMV-induced cross-protection. Each vaccinated group consisted of 12 mice, and the PBS group consisted of 10 mice. The cross-reactivity data represent the exact concentration of total IgG antibodies in the sera, as quantified using the corresponding standard curve using individual sera obtained from mice immunized intranasally or intraperitoneally with OMVs derived from S. Typhimurium. The error bars represent variations between triplicate wells. Competitive ELISA to determine the cross-reactivity of OMVs derived from flagellin-deficient mutant against heterologous Salmonella. OMPs isolated from S. Typhimurium were incubated into plates as coating antigen. OMPs from S. Choleraesuis, S. Enteritidis or S. Typhimurium (control) as the competitive antigen diluted from 1/10 to 1/ 7,290 were incubated in wells. The sera were obtained from mice (n = 10 or 12) immunized intranasally ( c ) or intraperitoneally ( d ) with OMVs at 8 weeks after the first immunization. The error bars represent variations from triplicate wells. P

Techniques Used: Derivative Assay, Mutagenesis, Mouse Assay, Concentration Assay, Competitive ELISA, Isolation, Incubation

Characterization, visualization and cytotoxicity of OMVs derived from S. Typhimurium and non-flagellin mutant strain. ( a ) Cryo-EM imaging of OMVs. OMVs derived from the flagellin-deficient S. Typhimurium were visualized using cryo-EM. The red arrows indicate the visible OMVs. ( b ) In total, 10 μg of OMVs from each sample was subjected to 12% SDS-PAGE and stained with GelCode TM Blue Stain. The major OMPs, including OmpA, OmpC/F and OmpD, are marked on the left, and the flagellar FliC and FljB proteins are labelled on the right. ( c ) LPS profiles from OMVs. LPS obtained from OMVs was visualized using silver staining after the samples were separated using 12% SDS-PAGE. ( d ) Quantification of LPS levels in OMVs. The same amount of OMVs (50 μg) was measured using a Kdo (3-deoxy-D-manno-octulosonic acid) analysis. S. Typhimurium LPS was used as the standard. ( e ) The cytotoxicity of OMVs derived from S. Typhimurium and the flagellin-deficient mutant in RAW264.7 macrophage cells. Cells were incubated with the corresponding OMVs at the indicated dose. Cell viability was determined by measuring the fluorescence in the supernatants using a Multitox-Fluor Multiplex Cytotoxicity Assay. Supernatants from cells without OMVs and cell lysis solution were treated to induce cell lysis, and these products were used as the negative and positive controls, respectively. Two-way ANOVA was performed to determine the significance of differences.
Figure Legend Snippet: Characterization, visualization and cytotoxicity of OMVs derived from S. Typhimurium and non-flagellin mutant strain. ( a ) Cryo-EM imaging of OMVs. OMVs derived from the flagellin-deficient S. Typhimurium were visualized using cryo-EM. The red arrows indicate the visible OMVs. ( b ) In total, 10 μg of OMVs from each sample was subjected to 12% SDS-PAGE and stained with GelCode TM Blue Stain. The major OMPs, including OmpA, OmpC/F and OmpD, are marked on the left, and the flagellar FliC and FljB proteins are labelled on the right. ( c ) LPS profiles from OMVs. LPS obtained from OMVs was visualized using silver staining after the samples were separated using 12% SDS-PAGE. ( d ) Quantification of LPS levels in OMVs. The same amount of OMVs (50 μg) was measured using a Kdo (3-deoxy-D-manno-octulosonic acid) analysis. S. Typhimurium LPS was used as the standard. ( e ) The cytotoxicity of OMVs derived from S. Typhimurium and the flagellin-deficient mutant in RAW264.7 macrophage cells. Cells were incubated with the corresponding OMVs at the indicated dose. Cell viability was determined by measuring the fluorescence in the supernatants using a Multitox-Fluor Multiplex Cytotoxicity Assay. Supernatants from cells without OMVs and cell lysis solution were treated to induce cell lysis, and these products were used as the negative and positive controls, respectively. Two-way ANOVA was performed to determine the significance of differences.

Techniques Used: Derivative Assay, Mutagenesis, Imaging, SDS Page, Staining, Silver Staining, Incubation, Fluorescence, Multiplex Assay, Cytotoxicity Assay, Lysis

Cross-protection of OMVs derived from flagellin-deficient S. Typhimurium mutant strain. Immunized mice were challenged orally with 10 7 CFU (~100-fold LD 50 ) or 10 7 CFU (~100-fold LD 50 ) of wild-type S. Choleraesuis or S. Enteritidis, respectively. Mortality was monitored for 3 weeks after challenge. ( a ) Survival was followed in mice that were intranasally immunized using OMVs and subsequently submitted to by S. Choleraesuis challenge. ( b ) Survival was followed in mice that were intranasally immunized with OMVs and subsequently submitted to S. Enteritidis challenge. ( c ) Survival was followed in mice that were intraperitoneally immunized with OMVs and then submitted S. Choleraesuis challenge. ( d ) Survival was followed in mice that were intraperitoneally immunized with OMVs and then submitted to S. Enteritidis challenge. All vaccine groups were significantly different from the PBS control group ( P
Figure Legend Snippet: Cross-protection of OMVs derived from flagellin-deficient S. Typhimurium mutant strain. Immunized mice were challenged orally with 10 7 CFU (~100-fold LD 50 ) or 10 7 CFU (~100-fold LD 50 ) of wild-type S. Choleraesuis or S. Enteritidis, respectively. Mortality was monitored for 3 weeks after challenge. ( a ) Survival was followed in mice that were intranasally immunized using OMVs and subsequently submitted to by S. Choleraesuis challenge. ( b ) Survival was followed in mice that were intranasally immunized with OMVs and subsequently submitted to S. Enteritidis challenge. ( c ) Survival was followed in mice that were intraperitoneally immunized with OMVs and then submitted S. Choleraesuis challenge. ( d ) Survival was followed in mice that were intraperitoneally immunized with OMVs and then submitted to S. Enteritidis challenge. All vaccine groups were significantly different from the PBS control group ( P

Techniques Used: Derivative Assay, Mutagenesis, Mouse Assay

5) Product Images from "Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿"

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿

Journal: Infection and Immunity

doi: 10.1128/IAI.00581-10

Time course of the S. Typhimurium binding to host cells. HeLa cells were infected with the indicated S. Typhimurium strains at an MOI of 62.5 for the indicated times followed by analysis of S. Typhimurium binding.
Figure Legend Snippet: Time course of the S. Typhimurium binding to host cells. HeLa cells were infected with the indicated S. Typhimurium strains at an MOI of 62.5 for the indicated times followed by analysis of S. Typhimurium binding.

Techniques Used: Binding Assay, Infection

Automated analysis of S. Typhimurium binding. (A) Image analysis strategy to quantify S. Typhimurium binding. HeLa cells were incubated with the Δ4 strain (MOI = 62.5) for 10 min at 37°C. (Upper left panel) Bacteria were stained by an anti- S. Typhimurium antibody (green), nuclei were stained with DAPI (gray), and the actin cytoskeleton was stained with TRITC-phalloidin (red). The following panels demonstrate image analysis by the open source program CellProfiler and custom algorithms. (Middle left panel) Recognition of nuclei. (Lower left panel) Expansion of the nuclear area to estimate the dimensions of cells. (Upper right panel) Overlay of the cell borders over the actin stain for illustration (the actin stain was not used for the image analysis). (Middle right panel) Detection of spots in the Salmonella channel using a threshold that is calculated by comparing infected and noninfected wells. (Lower right panel) Spots and cells are superimposed. Spots are allocated to the cell with the greatest overlap. Cells containing at least one spot are scored as having bound bacteria (red outlines), and cells without S. Typhimurium are scored as noninfected (blue outlines). Only a small part of one image is shown. In a typical well, 10,000 cells are identified within the four acquired images. The automated analysis for this well yielded a ratio of infected cells of 49.6%. Scale bar, 100 μm. (B) S. Typhimurium binding to cells via T1-dependent and -independent mechanisms. HeLa cells were infected with the different S. Typhimurium strains at the indicated MOIs and incubated for 12 min. The medians and standard deviations of five independent experiments are shown. An asterisk (*) indicates a P value below 0.05 (Mann-Whitney U test) comparing the tested data point to the Δ4 strain at the same MOI; an asterisk in parentheses indicates a P value below 0.05 comparing the T1 − Fi − strain to the T1 − strain. Points not marked by an asterisk of the same curve were also tested but did not yield significant P values. (C) Efficient S. Typhimurium binding requires an intact T1 system. HeLa cells were infected as in panel B using either the Δ4 strain or an isogenic strain carrying a combined knockout of sipB , sipC , and sipD (the SipBCD − strain). The T1 − strain and a mutant lacking the three translocases as well as T1 (the T1 − SipBCD − strain) are shown for comparison. An asterisk next to the curve indicates a P value below 0.05 (Wilcoxon signed-rank test) comparing the tested curve (median values) to the Δ4 strain.
Figure Legend Snippet: Automated analysis of S. Typhimurium binding. (A) Image analysis strategy to quantify S. Typhimurium binding. HeLa cells were incubated with the Δ4 strain (MOI = 62.5) for 10 min at 37°C. (Upper left panel) Bacteria were stained by an anti- S. Typhimurium antibody (green), nuclei were stained with DAPI (gray), and the actin cytoskeleton was stained with TRITC-phalloidin (red). The following panels demonstrate image analysis by the open source program CellProfiler and custom algorithms. (Middle left panel) Recognition of nuclei. (Lower left panel) Expansion of the nuclear area to estimate the dimensions of cells. (Upper right panel) Overlay of the cell borders over the actin stain for illustration (the actin stain was not used for the image analysis). (Middle right panel) Detection of spots in the Salmonella channel using a threshold that is calculated by comparing infected and noninfected wells. (Lower right panel) Spots and cells are superimposed. Spots are allocated to the cell with the greatest overlap. Cells containing at least one spot are scored as having bound bacteria (red outlines), and cells without S. Typhimurium are scored as noninfected (blue outlines). Only a small part of one image is shown. In a typical well, 10,000 cells are identified within the four acquired images. The automated analysis for this well yielded a ratio of infected cells of 49.6%. Scale bar, 100 μm. (B) S. Typhimurium binding to cells via T1-dependent and -independent mechanisms. HeLa cells were infected with the different S. Typhimurium strains at the indicated MOIs and incubated for 12 min. The medians and standard deviations of five independent experiments are shown. An asterisk (*) indicates a P value below 0.05 (Mann-Whitney U test) comparing the tested data point to the Δ4 strain at the same MOI; an asterisk in parentheses indicates a P value below 0.05 comparing the T1 − Fi − strain to the T1 − strain. Points not marked by an asterisk of the same curve were also tested but did not yield significant P values. (C) Efficient S. Typhimurium binding requires an intact T1 system. HeLa cells were infected as in panel B using either the Δ4 strain or an isogenic strain carrying a combined knockout of sipB , sipC , and sipD (the SipBCD − strain). The T1 − strain and a mutant lacking the three translocases as well as T1 (the T1 − SipBCD − strain) are shown for comparison. An asterisk next to the curve indicates a P value below 0.05 (Wilcoxon signed-rank test) comparing the tested curve (median values) to the Δ4 strain.

Techniques Used: Binding Assay, Incubation, Staining, Infection, MANN-WHITNEY, Knock-Out, Mutagenesis

Dissociation kinetics of T1- and Fim-mediated binding. The indicated S. Typhimurium strains were incubated with HeLa cells for 12 min. Plates were subsequently washed and incubated for 10 min at 37°C in medium containing α-methyl-mannose. Several rounds of medium exchange and incubation were performed. The appropriate wells were fixed. Finally, binding was measured as described in the text.
Figure Legend Snippet: Dissociation kinetics of T1- and Fim-mediated binding. The indicated S. Typhimurium strains were incubated with HeLa cells for 12 min. Plates were subsequently washed and incubated for 10 min at 37°C in medium containing α-methyl-mannose. Several rounds of medium exchange and incubation were performed. The appropriate wells were fixed. Finally, binding was measured as described in the text.

Techniques Used: Binding Assay, Incubation

Fundamental characteristics of the automated S . Typhimurium binding assay. (A and B) Increasing numbers of HeLa cells were seeded and infected with the indicated S . Typhimurium strain for 12 min. S . Typhimurium binding was determined as described in the text and plotted as a function of the number of nuclei detected within the respective well. The data represent the medians of three independent experiments and the standard deviations. (C) Comparison of S . Typhimurium binding to HeLa cells and “empty” areas of the same well. The spot density of an empty area of a well was calculated and expressed as a fraction of the spot density of the cell area of the same well. The data represent the medians and standard deviations of 87 to 90 wells at various MOIs from three independent experiments.
Figure Legend Snippet: Fundamental characteristics of the automated S . Typhimurium binding assay. (A and B) Increasing numbers of HeLa cells were seeded and infected with the indicated S . Typhimurium strain for 12 min. S . Typhimurium binding was determined as described in the text and plotted as a function of the number of nuclei detected within the respective well. The data represent the medians of three independent experiments and the standard deviations. (C) Comparison of S . Typhimurium binding to HeLa cells and “empty” areas of the same well. The spot density of an empty area of a well was calculated and expressed as a fraction of the spot density of the cell area of the same well. The data represent the medians and standard deviations of 87 to 90 wells at various MOIs from three independent experiments.

Techniques Used: Binding Assay, Infection

Role of type I fimbriae for S. Typhimurium binding to HeLa cells. HeLa cells were infected with the indicated S. Typhimurium strains for 12 min, and binding was analyzed. (A) Deletion mutants of type I fimbriae, shdA , long polar fimbriae, plasmid-encoded fimbriae, and thin aggregative fimbriae were analyzed in the background of the T1 − mutant. (B) Deletion mutants of the fim operon were generated in the background of the T1 − strain ( fimA , fimD , and fimH ), the Δ4 strain ( fimD , fimA , and fimH ), and the wild-type strain ( fimD) , respectively, and tested for HeLa cell binding. An asterisk next to the curve indicates a P value below 0.05 (Wilcoxon signed-rank test) comparing the indicated curve (median values) to the T1 − strain.
Figure Legend Snippet: Role of type I fimbriae for S. Typhimurium binding to HeLa cells. HeLa cells were infected with the indicated S. Typhimurium strains for 12 min, and binding was analyzed. (A) Deletion mutants of type I fimbriae, shdA , long polar fimbriae, plasmid-encoded fimbriae, and thin aggregative fimbriae were analyzed in the background of the T1 − mutant. (B) Deletion mutants of the fim operon were generated in the background of the T1 − strain ( fimA , fimD , and fimH ), the Δ4 strain ( fimD , fimA , and fimH ), and the wild-type strain ( fimD) , respectively, and tested for HeLa cell binding. An asterisk next to the curve indicates a P value below 0.05 (Wilcoxon signed-rank test) comparing the indicated curve (median values) to the T1 − strain.

Techniques Used: Binding Assay, Infection, Plasmid Preparation, Mutagenesis, Generated

Bound S. Typhimurium can proceed with cellular invasion if ruffling is triggered in trans . (Left panel) Scheme of the helper assay. S. Typhimurium carrying plasmid pM975 for intracellular GFP production was allowed to bind, followed by washing and addition of the helper strain. The helper strain induces ruffling and internalization of both the previously bound S. Typhimurium and the helper bacteria. S. Typhimurium was allowed to express gfp during another 4 h in medium containing gentamicin for killing extracellular S. Typhimurium. Binding of the Δ4 strain and the T1 − strain (black symbols) was evaluated by anti-LPS staining and automated microscopy. (Middle panel) Microscopy images showing induction of gfp expression by the Δ4 strain (pM975) with the wild-type strain as the helper and no gfp expression with the Δ4 strain as a helper strain. Scale bar, 100 μm. (Right panel) Quantification of binding at an MOI of 125 for various time points (black dashed lines) and invasion (red lines) of the indicated combinations of noninvasive S. Typhimurium and helpers. The helper was tested at various concentrations, and the maximum invasion for each concentration was plotted. For curves marked by two asterisks, a P value of less than 0.001 was obtained when invasion in the presence of the wt strain as a helper was compared to the Δ4 strain or the T1 − strain. For these analyses, all values of the three biological replicas were used in a paired test (Wilcoxon signed-rank test).
Figure Legend Snippet: Bound S. Typhimurium can proceed with cellular invasion if ruffling is triggered in trans . (Left panel) Scheme of the helper assay. S. Typhimurium carrying plasmid pM975 for intracellular GFP production was allowed to bind, followed by washing and addition of the helper strain. The helper strain induces ruffling and internalization of both the previously bound S. Typhimurium and the helper bacteria. S. Typhimurium was allowed to express gfp during another 4 h in medium containing gentamicin for killing extracellular S. Typhimurium. Binding of the Δ4 strain and the T1 − strain (black symbols) was evaluated by anti-LPS staining and automated microscopy. (Middle panel) Microscopy images showing induction of gfp expression by the Δ4 strain (pM975) with the wild-type strain as the helper and no gfp expression with the Δ4 strain as a helper strain. Scale bar, 100 μm. (Right panel) Quantification of binding at an MOI of 125 for various time points (black dashed lines) and invasion (red lines) of the indicated combinations of noninvasive S. Typhimurium and helpers. The helper was tested at various concentrations, and the maximum invasion for each concentration was plotted. For curves marked by two asterisks, a P value of less than 0.001 was obtained when invasion in the presence of the wt strain as a helper was compared to the Δ4 strain or the T1 − strain. For these analyses, all values of the three biological replicas were used in a paired test (Wilcoxon signed-rank test).

Techniques Used: Plasmid Preparation, Binding Assay, Staining, Microscopy, Expressing, Concentration Assay

6) Product Images from "Salmonella enterica serotype Typhimurium Std fimbriae bind terminal ? (1,2)fucose residues in the cecal mucosa"

Article Title: Salmonella enterica serotype Typhimurium Std fimbriae bind terminal ? (1,2)fucose residues in the cecal mucosa

Journal: Molecular microbiology

doi: 10.1111/j.1365-2958.2008.06566.x

Binding of S. Typhimurium to live Caco-2 cells. (A) S. Typhimurium strains were allowed to adhere to Caco-2 cells at 4°C in the absence (black bars) or presence of Galβ1-4GlcNAc (Gray bars). (B) S. Typhimurium strains were allowed to adhere to Caco-2 cells treated with α(1,2)fucosidase (gray bars) or to untreated Caco-2 cells (black bars) at 4°C. (C) S. Typhimurium strains were allowed to adhere to Caco-2 cells treated with neuraminidase (gray bars) or to untreated Caco-2 cells (black bars) at 4°C. (D) S. Typhimurium strains were allowed to adhere to Caco-2 cells at 4°C in the absence (black bars) or presence of α-L-fucose-PAA (Gray bars). All data are shown as averages of cell associated bacteria ± standard error from four independent experiments. Statistical significance of differences is indicated by brackets.
Figure Legend Snippet: Binding of S. Typhimurium to live Caco-2 cells. (A) S. Typhimurium strains were allowed to adhere to Caco-2 cells at 4°C in the absence (black bars) or presence of Galβ1-4GlcNAc (Gray bars). (B) S. Typhimurium strains were allowed to adhere to Caco-2 cells treated with α(1,2)fucosidase (gray bars) or to untreated Caco-2 cells (black bars) at 4°C. (C) S. Typhimurium strains were allowed to adhere to Caco-2 cells treated with neuraminidase (gray bars) or to untreated Caco-2 cells (black bars) at 4°C. (D) S. Typhimurium strains were allowed to adhere to Caco-2 cells at 4°C in the absence (black bars) or presence of α-L-fucose-PAA (Gray bars). All data are shown as averages of cell associated bacteria ± standard error from four independent experiments. Statistical significance of differences is indicated by brackets.

Techniques Used: Binding Assay

Binding of S. Typhimurium (A) and S. Typhi (B) to live human colonic epithelial cells. (A) S. Typhimurium strains were allowed to adhere to Caco-2 cells treated with peptide N-glycosidase F (gray bars) or to untreated Caco-2 cells (black bars) at 4°C to prevent bacterial invasion. Gentamicin treatment of bacteria attached to Caco-2 cells at 4°C was performed as a control (open bars). (B) S. Typhi strains were allowed to adhere to Caco-2 cells (black bars) or T84 cells (gray bars) at 4°C to prevent bacterial invasion. All data are shown as averages of cell associated bacteria ± standard error from four independent experiments. Statistical significance of differences is indicated by brackets. (C) Expression of StdA detected by Western blot in S. Typhi with anti-StdA serum.
Figure Legend Snippet: Binding of S. Typhimurium (A) and S. Typhi (B) to live human colonic epithelial cells. (A) S. Typhimurium strains were allowed to adhere to Caco-2 cells treated with peptide N-glycosidase F (gray bars) or to untreated Caco-2 cells (black bars) at 4°C to prevent bacterial invasion. Gentamicin treatment of bacteria attached to Caco-2 cells at 4°C was performed as a control (open bars). (B) S. Typhi strains were allowed to adhere to Caco-2 cells (black bars) or T84 cells (gray bars) at 4°C to prevent bacterial invasion. All data are shown as averages of cell associated bacteria ± standard error from four independent experiments. Statistical significance of differences is indicated by brackets. (C) Expression of StdA detected by Western blot in S. Typhi with anti-StdA serum.

Techniques Used: Binding Assay, Expressing, Western Blot

Binding of S. Typhimurium to live Caco-2 cells in the presence of soluble carbohydrates. (A) S. Typhimurium strains were allowed to adhere to Caco-2 cells at 4°C in the absence (black bars) or presence of soluble carbohydrates (Gray bars), including H type 2 or D mannose. All data are shown as averages of cell associated bacteria ± standard error from four independent experiments. Statistical significance of differences is indicated by brackets. (B) Expression of StdA detected by Western blot in cultures used in the adherence assays shown above.
Figure Legend Snippet: Binding of S. Typhimurium to live Caco-2 cells in the presence of soluble carbohydrates. (A) S. Typhimurium strains were allowed to adhere to Caco-2 cells at 4°C in the absence (black bars) or presence of soluble carbohydrates (Gray bars), including H type 2 or D mannose. All data are shown as averages of cell associated bacteria ± standard error from four independent experiments. Statistical significance of differences is indicated by brackets. (B) Expression of StdA detected by Western blot in cultures used in the adherence assays shown above.

Techniques Used: Binding Assay, Expressing, Western Blot

Binding of S. Typhimurium to formalin fixed Caco-2 cells. (A) Expression of StdA detected by Western blot using rabbit anti-StdA serum. (B) Expression of StdA on the surface of S. Typhimurium was detected by flow cytometry with rabbit anti-StdA Alexa Flour 647 conjugate (y axes). (C) Visualization of bacterial attachment to formalin fixed Caco-2 cells using Giemsa staining. (D) Binding of the S. Typhimurium fim mutant (open circles) and the Std fimbriated S. Typhimurium fim rosE mutant (closed circles) to formalin fixed Caco-2 cells was detected using rabbit anti-O4 serum and goat anti-rabbit alkaline phosphatase conjugate. Data are shown as average from three independent experiments ± standard error.
Figure Legend Snippet: Binding of S. Typhimurium to formalin fixed Caco-2 cells. (A) Expression of StdA detected by Western blot using rabbit anti-StdA serum. (B) Expression of StdA on the surface of S. Typhimurium was detected by flow cytometry with rabbit anti-StdA Alexa Flour 647 conjugate (y axes). (C) Visualization of bacterial attachment to formalin fixed Caco-2 cells using Giemsa staining. (D) Binding of the S. Typhimurium fim mutant (open circles) and the Std fimbriated S. Typhimurium fim rosE mutant (closed circles) to formalin fixed Caco-2 cells was detected using rabbit anti-O4 serum and goat anti-rabbit alkaline phosphatase conjugate. Data are shown as average from three independent experiments ± standard error.

Techniques Used: Binding Assay, Expressing, Western Blot, Flow Cytometry, Cytometry, Staining, Mutagenesis

7) Product Images from "Real-time imaging of type III secretion: Salmonella SipA injection into host cells"

Article Title: Real-time imaging of type III secretion: Salmonella SipA injection into host cells

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0503407102

Live imaging of SipA injection into host cells. ( A ) ( Left ) Time series showing GFP-InvB recruitment (arrows in GFP channel) after docking of S . typhimurium M1304 (arrows in phase contrast) to GFP-InvB-expressing Cos7 cells. White circles highlight bacteria that did not dock. t , time of recording. Bacteria were added at t = 240 sec. ( Center ) For t = 778 sec, all 5 z sections are shown for the region of interest. ( Right ) After recording ( t = 24 min), colocalization of GFP-InvB with SipA was verified. ( B ) Time between bacterial docking and first detection of GFP-InvB accumulation was analyzed from time-lapse recordings as shown in A . n , number of individual injection events analyzed in the indicated number of independent experiments. ( C ) Individual time courses of GFP-InvB recruitment. * , The number of transported SipA molecules was estimated as described in Supporting Materials and Methods after calibration of the imaging system. The time of bacterial docking to the cell was set to t = 0 (arrow) in the 10 curves obtained from six independent experiments. Dotted lines indicate the estimated average (21 sec -1 ), minimal (7 sec -1 ), and maximal (60 sec -1 ) rates of SipA injection observed. ( Inset ) Assignment of test (white) and background (gray) regions used for background correction and calculating GFP-InvB recruitment (see Supporting Materials and Methods ). Identical results were obtained with SipA M45 and native SipA (data not shown). ( D ) ( Lower ) SipA depletion from the bacterial cytosol was verified by fixing bacteria during ( Left ) or after ( Right ) completion of GFP-InvB recruitment. ( Upper ) Bacteria were lysozyme-permeabilized, and LPS (red; bacterial surface) and SipA (blue) in the bacterial and host cell cytosol were immunostained (see Materials and Methods ). GFP-InvB is shown in green. Images show confocal xyz sections of three-dimensional image reconstructions.
Figure Legend Snippet: Live imaging of SipA injection into host cells. ( A ) ( Left ) Time series showing GFP-InvB recruitment (arrows in GFP channel) after docking of S . typhimurium M1304 (arrows in phase contrast) to GFP-InvB-expressing Cos7 cells. White circles highlight bacteria that did not dock. t , time of recording. Bacteria were added at t = 240 sec. ( Center ) For t = 778 sec, all 5 z sections are shown for the region of interest. ( Right ) After recording ( t = 24 min), colocalization of GFP-InvB with SipA was verified. ( B ) Time between bacterial docking and first detection of GFP-InvB accumulation was analyzed from time-lapse recordings as shown in A . n , number of individual injection events analyzed in the indicated number of independent experiments. ( C ) Individual time courses of GFP-InvB recruitment. * , The number of transported SipA molecules was estimated as described in Supporting Materials and Methods after calibration of the imaging system. The time of bacterial docking to the cell was set to t = 0 (arrow) in the 10 curves obtained from six independent experiments. Dotted lines indicate the estimated average (21 sec -1 ), minimal (7 sec -1 ), and maximal (60 sec -1 ) rates of SipA injection observed. ( Inset ) Assignment of test (white) and background (gray) regions used for background correction and calculating GFP-InvB recruitment (see Supporting Materials and Methods ). Identical results were obtained with SipA M45 and native SipA (data not shown). ( D ) ( Lower ) SipA depletion from the bacterial cytosol was verified by fixing bacteria during ( Left ) or after ( Right ) completion of GFP-InvB recruitment. ( Upper ) Bacteria were lysozyme-permeabilized, and LPS (red; bacterial surface) and SipA (blue) in the bacterial and host cell cytosol were immunostained (see Materials and Methods ). GFP-InvB is shown in green. Images show confocal xyz sections of three-dimensional image reconstructions.

Techniques Used: Imaging, Injection, Expressing, Size-exclusion Chromatography

SipA expression by S . typhimurium and SipA localization in infected Cos7 cells. ( A ) Detection of SipA in the cytosol of individual bacteria from S . typhimurium cultures used for infection experiments (see Materials and Methods ). S . typhimurium M1300, M1304, and M1301 and control bacteria were immobilized and permeabilized by lysozyme treatment, and SipA in the bacterial cytosol was immunostained (see Fig. 5). SipA levels in ≥500 bacteria from three independent experiments were evaluated for each strain. ( B ) Average SipA levels of the cultures from A were determined by quantitative Western blotting and plating (see Fig. 5 A ). Data are expressed as the average number of SipA molecules per sipA -expressing bacterium. ( A and B ) Control indicates the background signal from bacteria expressing SipA without the M45 epítope tag. Error bars indicate standard deviation. ( C ) Localization of delivered SipA (red) in Cos7 cells infected for 30 min with wild-type S . typhimurium (M1300) or with the noninvasive strain M1304. Bacteria were stained before (green) and after (blue) permeabilization of the Cos7 cells to distinguish intracellular (blue) and extracellular (cyan in merge) bacteria. ( D ) Three-dimensional reconstruction of confocal image stacks showing SipA (red) delivered by individual bacteria (green; anti-LPS) of S . typhimurium strain M1300 or M1304 into a Cos7 cell 10 min after infection.
Figure Legend Snippet: SipA expression by S . typhimurium and SipA localization in infected Cos7 cells. ( A ) Detection of SipA in the cytosol of individual bacteria from S . typhimurium cultures used for infection experiments (see Materials and Methods ). S . typhimurium M1300, M1304, and M1301 and control bacteria were immobilized and permeabilized by lysozyme treatment, and SipA in the bacterial cytosol was immunostained (see Fig. 5). SipA levels in ≥500 bacteria from three independent experiments were evaluated for each strain. ( B ) Average SipA levels of the cultures from A were determined by quantitative Western blotting and plating (see Fig. 5 A ). Data are expressed as the average number of SipA molecules per sipA -expressing bacterium. ( A and B ) Control indicates the background signal from bacteria expressing SipA without the M45 epítope tag. Error bars indicate standard deviation. ( C ) Localization of delivered SipA (red) in Cos7 cells infected for 30 min with wild-type S . typhimurium (M1300) or with the noninvasive strain M1304. Bacteria were stained before (green) and after (blue) permeabilization of the Cos7 cells to distinguish intracellular (blue) and extracellular (cyan in merge) bacteria. ( D ) Three-dimensional reconstruction of confocal image stacks showing SipA (red) delivered by individual bacteria (green; anti-LPS) of S . typhimurium strain M1300 or M1304 into a Cos7 cell 10 min after infection.

Techniques Used: Expressing, Infection, Western Blot, Standard Deviation, Staining

GFP-InvB is recruited to SipA foci at the site of bacteria-host cell contact. ( A ) Schematic of the GFP-InvB recruitment assay for studying SipA delivery into host cells via the Salmonella SPI-1 TTSS. ( B and C ) Distribution of GFP-InvB (green) in uninfected Cos7 cells ( B ) and cells infected with S. typhimurium strain M712 ( C ), which lacks SipA. Bacteria were stained for LPS (blue). ( D ) GFP-InvB is recruited to foci of delivered SipA (magenta) after infection of pGFP-InvB-transfected cells with S. typhimurium strain M1304 (blue) for 30 min. SipA and recruited GFP-InvB colocalize with F-actin (red). ( E ) Specificity of GFP-InvB recruitment was verified by infecting Cos7 cells coexpressing pGFP-InvB and pmRFP-1 with M1304 (blue). GFP-InvB but not mRFP-1 (red) was recruited to SipA foci (magenta). ( F ) Bacteria lacking a functional SPI-1 TTSS (M1301) do not induce GFP-InvB recruitment.
Figure Legend Snippet: GFP-InvB is recruited to SipA foci at the site of bacteria-host cell contact. ( A ) Schematic of the GFP-InvB recruitment assay for studying SipA delivery into host cells via the Salmonella SPI-1 TTSS. ( B and C ) Distribution of GFP-InvB (green) in uninfected Cos7 cells ( B ) and cells infected with S. typhimurium strain M712 ( C ), which lacks SipA. Bacteria were stained for LPS (blue). ( D ) GFP-InvB is recruited to foci of delivered SipA (magenta) after infection of pGFP-InvB-transfected cells with S. typhimurium strain M1304 (blue) for 30 min. SipA and recruited GFP-InvB colocalize with F-actin (red). ( E ) Specificity of GFP-InvB recruitment was verified by infecting Cos7 cells coexpressing pGFP-InvB and pmRFP-1 with M1304 (blue). GFP-InvB but not mRFP-1 (red) was recruited to SipA foci (magenta). ( F ) Bacteria lacking a functional SPI-1 TTSS (M1301) do not induce GFP-InvB recruitment.

Techniques Used: Infection, Staining, Transfection, Functional Assay

Transport by the SPI-1 TTSS depletes intrabacterial SipA and SopE pools. ( A ) Strategy for detecting effector protein injection into host cells by monitoring the presence of SipA in the bacterial cytosol. ( B ) Representative images of S . typhimurium M1304 in the early, intermediate, or late phase of SipA injection into Cos7 cells. Cells were fixed, permeabilized with lysozyme, and LPS (blue) and SipA (red) were immunostained (see Materials and Methods ). ( C ) Time course of SipA depletion during the infection of Cos7 cells with S . typhimurium M1300 (wild type; black circles), M1304 (noninvasive; white circles), and M1301 (SPI-1 TTSS-disrupted; crosses). ( D ) Parallel detection of SopE and SipA depletion from the bacterial cytosol. Cos7 cells were infected for 30 min with wild-type S . typhimurium M1222, fixed, permeabilized with lysozyme, and immunostained for SopE (green) and SipA (red). The arrows point to a group of bacteria that had completed the SipA and SopE injection. Arrowheads indicate bacterium still harboring SopE and SipA in the cytosol. ( E ) Depletion of SopE and SipA from the bacterial cytosol. Infection was monitored by phase-contrast time-lapse microscopy. Cells were fixed and stained as in D . For each bacterium, the graph shows the time between docking and fixation, the location of SipA (as in B ), and the presence/absence of SopE in the bacterial cytosol. Lines connect SipA and SopE data from the same bacterium.
Figure Legend Snippet: Transport by the SPI-1 TTSS depletes intrabacterial SipA and SopE pools. ( A ) Strategy for detecting effector protein injection into host cells by monitoring the presence of SipA in the bacterial cytosol. ( B ) Representative images of S . typhimurium M1304 in the early, intermediate, or late phase of SipA injection into Cos7 cells. Cells were fixed, permeabilized with lysozyme, and LPS (blue) and SipA (red) were immunostained (see Materials and Methods ). ( C ) Time course of SipA depletion during the infection of Cos7 cells with S . typhimurium M1300 (wild type; black circles), M1304 (noninvasive; white circles), and M1301 (SPI-1 TTSS-disrupted; crosses). ( D ) Parallel detection of SopE and SipA depletion from the bacterial cytosol. Cos7 cells were infected for 30 min with wild-type S . typhimurium M1222, fixed, permeabilized with lysozyme, and immunostained for SopE (green) and SipA (red). The arrows point to a group of bacteria that had completed the SipA and SopE injection. Arrowheads indicate bacterium still harboring SopE and SipA in the cytosol. ( E ) Depletion of SopE and SipA from the bacterial cytosol. Infection was monitored by phase-contrast time-lapse microscopy. Cells were fixed and stained as in D . For each bacterium, the graph shows the time between docking and fixation, the location of SipA (as in B ), and the presence/absence of SopE in the bacterial cytosol. Lines connect SipA and SopE data from the same bacterium.

Techniques Used: Injection, Infection, Time-lapse Microscopy, Staining

8) Product Images from "A novel CsrA titration mechanism regulates fimbrial gene expression in Salmonella typhimurium"

Article Title: A novel CsrA titration mechanism regulates fimbrial gene expression in Salmonella typhimurium

Journal: The EMBO Journal

doi: 10.1038/emboj.2013.206

CsrA regulates expression of PefA. ( A ) Expression of FimA was detected in cell lysates of the indicated S. typhimurium strains using western blot. ( B ) Quantification of FimA levels in western blots ( N =3) by densitometry. ( C ) Relative expression of fimA
Figure Legend Snippet: CsrA regulates expression of PefA. ( A ) Expression of FimA was detected in cell lysates of the indicated S. typhimurium strains using western blot. ( B ) Quantification of FimA levels in western blots ( N =3) by densitometry. ( C ) Relative expression of fimA

Techniques Used: Expressing, Western Blot

SirA represses expression of PefA via downregulation of CsrB and CsrC. Expression of CsrB RNA ( A ) and CsrC RNA ( B ) was quantified by real-time PCR using RNA isolated from S. typhimurium wild type (wt) and the sirA mutant ( sirA ). Bars represent the average
Figure Legend Snippet: SirA represses expression of PefA via downregulation of CsrB and CsrC. Expression of CsrB RNA ( A ) and CsrC RNA ( B ) was quantified by real-time PCR using RNA isolated from S. typhimurium wild type (wt) and the sirA mutant ( sirA ). Bars represent the average

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Isolation, Mutagenesis

The CsrA-binding site in the 5′-UTR of the pefA transcript limits expression to host environments. Streptomycin pretreated mice were inoculated with the indicated S. typhimurium mutants and RNA was isolated from the inoculum cultures (inoculum)
Figure Legend Snippet: The CsrA-binding site in the 5′-UTR of the pefA transcript limits expression to host environments. Streptomycin pretreated mice were inoculated with the indicated S. typhimurium mutants and RNA was isolated from the inoculum cultures (inoculum)

Techniques Used: Binding Assay, Expressing, Mouse Assay, Isolation

Proposed model for the hierarchical control of fimbrial expression in S. typhimurium mediated by CsrA. The exceptionally high transcript levels of the fimAICDHF 5′-UTR can antagonize the regulatory effects of CsrA by binding and sequestering this
Figure Legend Snippet: Proposed model for the hierarchical control of fimbrial expression in S. typhimurium mediated by CsrA. The exceptionally high transcript levels of the fimAICDHF 5′-UTR can antagonize the regulatory effects of CsrA by binding and sequestering this

Techniques Used: Expressing, Binding Assay

9) Product Images from "Plasticity in structure and interactions is critical for the action of indolicidin, an antibacterial peptide of innate immune origin"

Article Title: Plasticity in structure and interactions is critical for the action of indolicidin, an antibacterial peptide of innate immune origin

Journal: Protein Science : A Publication of the Protein Society

doi:

The D-analogs of indolicidin and retro-indolicidin show similar functional behavior as the corresponding L-peptides. Comparison of the antibacterial activities against Salmonella typhimurium of the two peptides is plotted in a dose-dependent manner.
Figure Legend Snippet: The D-analogs of indolicidin and retro-indolicidin show similar functional behavior as the corresponding L-peptides. Comparison of the antibacterial activities against Salmonella typhimurium of the two peptides is plotted in a dose-dependent manner.

Techniques Used: Functional Assay

Comparison of indolicidin with retro-indolicidin. ( A ) Antibacterial activity against Salmonella typhimurium . ( B ) Dose-dependent effect of MgCl 2 on the antibacterial activity of the peptides against S. typhimurium . ( C ) Competitive displacement of dansyl polymyxin B in a dose-dependent manner by different peptides for endotoxin binding.
Figure Legend Snippet: Comparison of indolicidin with retro-indolicidin. ( A ) Antibacterial activity against Salmonella typhimurium . ( B ) Dose-dependent effect of MgCl 2 on the antibacterial activity of the peptides against S. typhimurium . ( C ) Competitive displacement of dansyl polymyxin B in a dose-dependent manner by different peptides for endotoxin binding.

Techniques Used: Activity Assay, Binding Assay

Comparison of tritrypticin analog Sym11 with indolicidin. ( A ) Antibacterial activity against Samonella typhimurium DVWYPYPYASGS is an unrelated peptide used as control. ( B ) Dose-dependent effect of MgCl 2 on the antibacterial activity of the peptides against S. typhimurium . ( C ) Competitive displacement of dansyl polymyxin B in a dose-dependent manner by different peptides for endotoxin binding. ( D ) Sequence comparisons involving Sym11, indolicidin (N-indo) and retro-indolicidin (R-indo).
Figure Legend Snippet: Comparison of tritrypticin analog Sym11 with indolicidin. ( A ) Antibacterial activity against Samonella typhimurium DVWYPYPYASGS is an unrelated peptide used as control. ( B ) Dose-dependent effect of MgCl 2 on the antibacterial activity of the peptides against S. typhimurium . ( C ) Competitive displacement of dansyl polymyxin B in a dose-dependent manner by different peptides for endotoxin binding. ( D ) Sequence comparisons involving Sym11, indolicidin (N-indo) and retro-indolicidin (R-indo).

Techniques Used: Activity Assay, Binding Assay, Sequencing

10) Product Images from "Repulsion and Metabolic Switches in the Collective Behavior of Bacterial Colonies"

Article Title: Repulsion and Metabolic Switches in the Collective Behavior of Bacterial Colonies

Journal: Biophysical Journal

doi: 10.1016/j.bpj.2009.04.018

The contours of arrest experimentally observed for two and four colonies of S. typhimurium and the theoretical prediction superimposed as full lines.
Figure Legend Snippet: The contours of arrest experimentally observed for two and four colonies of S. typhimurium and the theoretical prediction superimposed as full lines.

Techniques Used:

Dependency of the lag time on the initial bacterial density and nutrient concentration. ( a ) Distance traveled by the bacterial front in S. typhimurium colonies. ( Inset ) Data have been collapsed onto a single curve by a horizontal shift of −1 h
Figure Legend Snippet: Dependency of the lag time on the initial bacterial density and nutrient concentration. ( a ) Distance traveled by the bacterial front in S. typhimurium colonies. ( Inset ) Data have been collapsed onto a single curve by a horizontal shift of −1 h

Techniques Used: Concentration Assay

Repulsion versus merging of bacterial colonies. ( Upper row ) S. typhimurium at 30°C in medium A. ( a ) Repulsion with 0.4% glucose and 0.6% Eiken agar, after 24 h. ( b ) Merging with 0.05% glucose and 0.4% Eiken agar after 24 h. ( c ) Repulsion in P.
Figure Legend Snippet: Repulsion versus merging of bacterial colonies. ( Upper row ) S. typhimurium at 30°C in medium A. ( a ) Repulsion with 0.4% glucose and 0.6% Eiken agar, after 24 h. ( b ) Merging with 0.05% glucose and 0.4% Eiken agar after 24 h. ( c ) Repulsion in P.

Techniques Used:

11) Product Images from "Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry"

Article Title: Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.02397

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P

Techniques Used:

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P

Techniques Used:

12) Product Images from "Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry"

Article Title: Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.02397

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P

Techniques Used:

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P

Techniques Used:

13) Product Images from "Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry"

Article Title: Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.02397

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on cherry tomato achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗∗ P

Techniques Used:

Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P
Figure Legend Snippet: Reductions in aerobic bacteria (A) , E. coli (B) , mold (C) , yeast (D) , and S. Typhimurium (E) on strawberry achieved by different sanitizing methods. Data are expressed as mean ± SD ( n = 3). ∗ P

Techniques Used:

14) Product Images from "Antibacterial and Antioxidant Activity of Essential Oil Terpenes against Pathogenic and Spoilage-Forming Bacteria and Cell Structure-Activity Relationships Evaluated by SEM Microscopy"

Article Title: Antibacterial and Antioxidant Activity of Essential Oil Terpenes against Pathogenic and Spoilage-Forming Bacteria and Cell Structure-Activity Relationships Evaluated by SEM Microscopy

Journal: Molecules

doi: 10.3390/molecules191117773

Release of cellular material at 260-nm and 280-nm for E. coli O157:H7 ( A ); S . Typhimurium ( B ); and S. aureus ( C ) cells treated with combinations at FIC values. (T: α-Terpineol, L: Linalool, E: Eucalyptol, add: additive; syn: synergy).
Figure Legend Snippet: Release of cellular material at 260-nm and 280-nm for E. coli O157:H7 ( A ); S . Typhimurium ( B ); and S. aureus ( C ) cells treated with combinations at FIC values. (T: α-Terpineol, L: Linalool, E: Eucalyptol, add: additive; syn: synergy).

Techniques Used:

Inhibition of cells treated with ( A ) eucalyptol, S . Typhimurium; ( B ) linalool, S . Typhimurium; ( C ) eucalyptol, E. coli O157:H7; ( D ) linalool, E. coli O157:H7; ( E ) α-terpineol, S. aureus ; ( F ) eucalyptol, S. aureus ; ( G ) linalool, S. aureus ; ( H ) α-terpineol, S . Typhimurium. (T: α-Terpineol, L: Linalool, E: Eucalyptol).
Figure Legend Snippet: Inhibition of cells treated with ( A ) eucalyptol, S . Typhimurium; ( B ) linalool, S . Typhimurium; ( C ) eucalyptol, E. coli O157:H7; ( D ) linalool, E. coli O157:H7; ( E ) α-terpineol, S. aureus ; ( F ) eucalyptol, S. aureus ; ( G ) linalool, S. aureus ; ( H ) α-terpineol, S . Typhimurium. (T: α-Terpineol, L: Linalool, E: Eucalyptol).

Techniques Used: Inhibition

15) Product Images from "Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)"

Article Title: Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.00006-19

Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on sample surfaces or in washing solution (tap water [TW] plus 0.1% Tween 20) treated with the Spindle in
Figure Legend Snippet: Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on sample surfaces or in washing solution (tap water [TW] plus 0.1% Tween 20) treated with the Spindle in

Techniques Used:

Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on the sample surface or in washing solution (tap water [TW]) treated with the Spindle in combination with
Figure Legend Snippet: Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on the sample surface or in washing solution (tap water [TW]) treated with the Spindle in combination with

Techniques Used:

16) Product Images from "Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)"

Article Title: Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.00006-19

Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on sample surfaces or in washing solution (tap water [TW] plus 0.1% Tween 20) treated with the Spindle in
Figure Legend Snippet: Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on sample surfaces or in washing solution (tap water [TW] plus 0.1% Tween 20) treated with the Spindle in

Techniques Used:

Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on the sample surface or in washing solution (tap water [TW]) treated with the Spindle in combination with
Figure Legend Snippet: Surviving populations (log CFU/sample or log CFU/250 ml) of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on the sample surface or in washing solution (tap water [TW]) treated with the Spindle in combination with

Techniques Used:

17) Product Images from "Evaluation of Near-Infrared Pasteurization in Controlling Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Ready-To-Eat Sliced Ham"

Article Title: Evaluation of Near-Infrared Pasteurization in Controlling Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Ready-To-Eat Sliced Ham

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.00942-12

Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes inside two contiguous ham slices treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.
Figure Legend Snippet: Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes inside two contiguous ham slices treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.

Techniques Used:

Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes on ham slice surfaces treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.
Figure Legend Snippet: Survival curves of Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes on ham slice surfaces treated with NIR or conventional convective heating. The error bars indicate standard deviations calculated from triplicates.

Techniques Used:

18) Product Images from "InvF Is Required for Expression of Genes Encoding Proteins Secreted by the SPI1 Type III Secretion Apparatus in Salmonella typhimurium"

Article Title: InvF Is Required for Expression of Genes Encoding Proteins Secreted by the SPI1 Type III Secretion Apparatus in Salmonella typhimurium

Journal: Journal of Bacteriology

doi:

Supernatant proteins from wild-type and Δ invF strains. Lane 1, supernatant proteins from the secretion defective spaS mutant SVM514; lanes 2 to 7, S. typhimurium SL1344 containing pWSK130, pHD9 ( invF + ), and p hilA (lanes 2 to 4) and the invF mutant SVM579 containing the same plasmids in the same order (lanes 5 to 7); lanes 8 and 9, supernatant proteins from SVM579 strains containing the vector pVLT33 (lane 8) and the IPTG-inducible sigDE clone pHH37 (lane 9). Positions of molecular weight standards are indicated in kilodaltons on the left; previously identified secreted proteins are indicated on the right. Question marks denote proteins that have not been confirmed by immunoblot analysis. Proteins were prepared and analyzed as described in Materials and Methods.
Figure Legend Snippet: Supernatant proteins from wild-type and Δ invF strains. Lane 1, supernatant proteins from the secretion defective spaS mutant SVM514; lanes 2 to 7, S. typhimurium SL1344 containing pWSK130, pHD9 ( invF + ), and p hilA (lanes 2 to 4) and the invF mutant SVM579 containing the same plasmids in the same order (lanes 5 to 7); lanes 8 and 9, supernatant proteins from SVM579 strains containing the vector pVLT33 (lane 8) and the IPTG-inducible sigDE clone pHH37 (lane 9). Positions of molecular weight standards are indicated in kilodaltons on the left; previously identified secreted proteins are indicated on the right. Question marks denote proteins that have not been confirmed by immunoblot analysis. Proteins were prepared and analyzed as described in Materials and Methods.

Techniques Used: Mutagenesis, Plasmid Preparation, Molecular Weight

Model for the regulation of invasion/virulence gene expression in S. typhimurium . The direction of transcription for each gene cluster is indicated by closed arrows; open arrows represent putative transcripts of the inv-spa and sip/ssp genes. Question marks indicate either unidentified regulatory factors or unclear relationships between the designated regulator and the noted promoter.
Figure Legend Snippet: Model for the regulation of invasion/virulence gene expression in S. typhimurium . The direction of transcription for each gene cluster is indicated by closed arrows; open arrows represent putative transcripts of the inv-spa and sip/ssp genes. Question marks indicate either unidentified regulatory factors or unclear relationships between the designated regulator and the noted promoter.

Techniques Used: Expressing

19) Product Images from "Contribution of Salmonella typhimurium Virulence Factors to Diarrheal Disease in Calves"

Article Title: Contribution of Salmonella typhimurium Virulence Factors to Diarrheal Disease in Calves

Journal: Infection and Immunity

doi:

Histopathology of Peyer’s patches and ileum from perinatal calves inoculated per os with 10 10 CFU of the indicated strains of S. typhimurium per animal, photographed at a 40× magnification. Shown are hematoxylin-and-eosin-stained sections of Peyer’s patch (left column) and terminal ileum (right column). Short arrows indicate areas of marked lymphoid depletion; long arrows indicate zones of variable degrees of fibrinopurulent necrotizing ileitis at the mucosal surface.
Figure Legend Snippet: Histopathology of Peyer’s patches and ileum from perinatal calves inoculated per os with 10 10 CFU of the indicated strains of S. typhimurium per animal, photographed at a 40× magnification. Shown are hematoxylin-and-eosin-stained sections of Peyer’s patch (left column) and terminal ileum (right column). Short arrows indicate areas of marked lymphoid depletion; long arrows indicate zones of variable degrees of fibrinopurulent necrotizing ileitis at the mucosal surface.

Techniques Used: Histopathology, Staining

Representative examples of the gross pathology of Peyer’s patch and mucosa of the terminal ileum of calves inoculated orally with 10 10 CFU of different S. typhimurium strains. (A) Severe acute fibrinopurulent necrotizing enteritis with segmental or continuous pseudomembrane formation of calves inoculated with wild type (IR715), strain STN272 ( spvR ), or strain SVM255 ( sigD ). (B) Moderate to marked subacute fibrinopurulent enteritis often confined to Peyer’s patches of calves inoculated with STN119 ( spiB ) or STN166 ( rfaJ ). (C) Normal Peyer’s patch and ileal mucosa of calves inoculated with STN61 ( hilA ), STN162 ( prgH ), or CL1509 ( aroA ) or of uninoculated controls. Bar = 1 cm.
Figure Legend Snippet: Representative examples of the gross pathology of Peyer’s patch and mucosa of the terminal ileum of calves inoculated orally with 10 10 CFU of different S. typhimurium strains. (A) Severe acute fibrinopurulent necrotizing enteritis with segmental or continuous pseudomembrane formation of calves inoculated with wild type (IR715), strain STN272 ( spvR ), or strain SVM255 ( sigD ). (B) Moderate to marked subacute fibrinopurulent enteritis often confined to Peyer’s patches of calves inoculated with STN119 ( spiB ) or STN166 ( rfaJ ). (C) Normal Peyer’s patch and ileal mucosa of calves inoculated with STN61 ( hilA ), STN162 ( prgH ), or CL1509 ( aroA ) or of uninoculated controls. Bar = 1 cm.

Techniques Used:

20) Product Images from "In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella"

Article Title: In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1093/ecam/nep105

Importance of PGPE against S. typhimurium infection.
Figure Legend Snippet: Importance of PGPE against S. typhimurium infection.

Techniques Used: Infection

21) Product Images from "Cytotoxic T cell adjuvant effects of three Salmonella enterica flagellins"

Article Title: Cytotoxic T cell adjuvant effects of three Salmonella enterica flagellins

Journal: Brazilian Journal of Microbiology

doi: 10.1590/S1517-838220080001000011

Extraction of native flagellins from Salmonella enterica strains. Protein samples were sorted in polyacrylamide gels were stained with Comassie Blue (A) orcon in Western blots (B) with anti-flagellin antibodies. Samples: MW- molecular weight markers; lane 1: FliC i flagellin harvested from S . Typhimurium; lane 2: FljB flagellin harvested from S . Typhimurium; lane 3: FliC d flagellin harvested from S . Dublin SL5930 strain.
Figure Legend Snippet: Extraction of native flagellins from Salmonella enterica strains. Protein samples were sorted in polyacrylamide gels were stained with Comassie Blue (A) orcon in Western blots (B) with anti-flagellin antibodies. Samples: MW- molecular weight markers; lane 1: FliC i flagellin harvested from S . Typhimurium; lane 2: FljB flagellin harvested from S . Typhimurium; lane 3: FliC d flagellin harvested from S . Dublin SL5930 strain.

Techniques Used: Staining, Western Blot, Molecular Weight

22) Product Images from "The role of lipopolysaccharide injected systemically in the reactivation of collagen-induced arthritis in mice"

Article Title: The role of lipopolysaccharide injected systemically in the reactivation of collagen-induced arthritis in mice

Journal: British Journal of Pharmacology

doi: 10.1038/sj.bjp.0703166

Reactivation of CIA by varying types of LPS and lipid A. Mice were immunized with CII on day 0 followed by a booster injection on day 21 as described in Methods. On day 50, saline, 5 μg of LPS from E. coli, S. enteritidis, S. typhimurium , and K. peumoniae , and 2 μg of lipid A from E. coli were i.p. injected. The severity of arthritis was determined on day 50, i.e. immediately before administration of LPS and on day 55. Bars show the mean±s.e.mean of eight mice. * P
Figure Legend Snippet: Reactivation of CIA by varying types of LPS and lipid A. Mice were immunized with CII on day 0 followed by a booster injection on day 21 as described in Methods. On day 50, saline, 5 μg of LPS from E. coli, S. enteritidis, S. typhimurium , and K. peumoniae , and 2 μg of lipid A from E. coli were i.p. injected. The severity of arthritis was determined on day 50, i.e. immediately before administration of LPS and on day 55. Bars show the mean±s.e.mean of eight mice. * P

Techniques Used: Mouse Assay, Injection

23) Product Images from "The role of lipopolysaccharide injected systemically in the reactivation of collagen-induced arthritis in mice"

Article Title: The role of lipopolysaccharide injected systemically in the reactivation of collagen-induced arthritis in mice

Journal: British Journal of Pharmacology

doi: 10.1038/sj.bjp.0703166

Reactivation of CIA by varying types of LPS and lipid A. Mice were immunized with CII on day 0 followed by a booster injection on day 21 as described in Methods. On day 50, saline, 5 μg of LPS from E. coli, S. enteritidis, S. typhimurium , and K. peumoniae , and 2 μg of lipid A from E. coli were i.p. injected. The severity of arthritis was determined on day 50, i.e. immediately before administration of LPS and on day 55. Bars show the mean±s.e.mean of eight mice. * P
Figure Legend Snippet: Reactivation of CIA by varying types of LPS and lipid A. Mice were immunized with CII on day 0 followed by a booster injection on day 21 as described in Methods. On day 50, saline, 5 μg of LPS from E. coli, S. enteritidis, S. typhimurium , and K. peumoniae , and 2 μg of lipid A from E. coli were i.p. injected. The severity of arthritis was determined on day 50, i.e. immediately before administration of LPS and on day 55. Bars show the mean±s.e.mean of eight mice. * P

Techniques Used: Mouse Assay, Injection

24) Product Images from "The role of lipopolysaccharide injected systemically in the reactivation of collagen-induced arthritis in mice"

Article Title: The role of lipopolysaccharide injected systemically in the reactivation of collagen-induced arthritis in mice

Journal: British Journal of Pharmacology

doi: 10.1038/sj.bjp.0703166

Reactivation of CIA by varying types of LPS and lipid A. Mice were immunized with CII on day 0 followed by a booster injection on day 21 as described in Methods. On day 50, saline, 5 μg of LPS from E. coli, S. enteritidis, S. typhimurium , and K. peumoniae , and 2 μg of lipid A from E. coli were i.p. injected. The severity of arthritis was determined on day 50, i.e. immediately before administration of LPS and on day 55. Bars show the mean±s.e.mean of eight mice. * P
Figure Legend Snippet: Reactivation of CIA by varying types of LPS and lipid A. Mice were immunized with CII on day 0 followed by a booster injection on day 21 as described in Methods. On day 50, saline, 5 μg of LPS from E. coli, S. enteritidis, S. typhimurium , and K. peumoniae , and 2 μg of lipid A from E. coli were i.p. injected. The severity of arthritis was determined on day 50, i.e. immediately before administration of LPS and on day 55. Bars show the mean±s.e.mean of eight mice. * P

Techniques Used: Mouse Assay, Injection

25) Product Images from "Effect of atmospheric pressure plasma jet on the foodborne pathogens attached to commercial food containers"

Article Title: Effect of atmospheric pressure plasma jet on the foodborne pathogens attached to commercial food containers

Journal: Journal of Food Science and Technology

doi: 10.1007/s13197-015-2003-0

Salmonella Typhimurium counts (log CFU/cm 2 ) in the biofilms treated with atmospheric pressure plasma jet for 0, 5, and 10 min. a Collagen casing, b Polyethylene terephthalate, c Polypropylene. ( a-c Values with different letters within the same
Figure Legend Snippet: Salmonella Typhimurium counts (log CFU/cm 2 ) in the biofilms treated with atmospheric pressure plasma jet for 0, 5, and 10 min. a Collagen casing, b Polyethylene terephthalate, c Polypropylene. ( a-c Values with different letters within the same

Techniques Used:

Related Articles

Infection:

Article Title: Real-time imaging of type III secretion: Salmonella SipA injection into host cells
Article Snippet: Cells were infected for the indicated time with S. typhimurium at a multiplicity of infection of 25 bacteria per cell, fixed with 4% paraformaldehyde in PBS/4% sucrose for 20 min at 22°C, and permeabilized in 0.1% Triton X-100 for 5 min (see Figs. , , and ). .. S. typhimurium was stained with a rabbit anti- Salmonella O -1,4,5,12 ( ) antiserum (Difco) and goat anti-rabbit-FITC , anti-rabbit-7-amino-4-methylcoumarin-3-acetic acid (see ) or anti-rabbit-rhodamine (see Figs. and ) conjugate.

Article Title: Contribution of Salmonella typhimurium Virulence Factors to Diarrheal Disease in Calves
Article Snippet: A previous report indicates that in calves which survive an S. typhimurium infection diarrhea does not persist beyond the 10th day postinfection ( ). .. Shedding of S. typhimurium was monitored by taking daily fecal swabs, subsequently enriching them in tetrathionate broth (Difco), and plating them on brilliant green agar (BBL).

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿
Article Snippet: After the indicated time of infection, the plates were emptied and washed three times with 50 μl of DMEM, 10% FCS using a Wellmate (Matrix). .. S. Typhimurium was visualized by indirect immunofluorescence using a rabbit anti- S. Typhimurium antibody (Difco) and a fluorescein-5-isothiocyanate (FITC)-labeled goat anti-rabbit secondary antibody (Jackson).

Centrifugation:

Article Title: Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)
Article Snippet: .. A single colony of each strain of E. coli O157:H7, S. Typhimurium, and L. monocytogenes cultured from stocks on tryptic soy agar (TSA) (Difco, Becton, Dickinson, Sparks, MD), were inoculated individually into 15 ml tryptic soy broth (TSB) (Difco), incubated at 37°C for 24 h, and then collected by centrifugation at 4,000 × g for 20 min at 4°C. .. After three washes with 0.2% peptone water (PW) (Bacto, Becton, Dickinson, Sparks, MD), suspended pellets of the three pathogens were combined to constitute a mixed-pathogen-species culture cocktail corresponding to approximately 108 to 109 CFU/ml.

Article Title: Evaluation of Near-Infrared Pasteurization in Controlling Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Ready-To-Eat Sliced Ham
Article Snippet: .. Each strain of S. Typhimurium, E. coli O157:H7, and L. monocytogenes was cultured in 5 ml of TSB at 37°C for 24 h, followed by centrifugation (4,000 × g for 20 min at 4°C) and washing three times with buffered peptone water (BPW; Difco, Sparks, MD). ..

Agglutination:

Article Title: Effects of mannoprotein E1 in liquid diet on inflammatory response and TLR5 expression in the gut of rats infected by Salmonella typhimurium
Article Snippet: .. Colony identification was made its morphological characteristics and agglutination with specific anti-sera to Salmonella poly A and S. typhimurium (Difco). .. Samples from the ileum, jejunum and colon were taken and placed in 10% neutral buffered formalin during 24 hr for Q-PCR and Western-blot assay.

Mutagenesis:

Article Title: Plasticity in structure and interactions is critical for the action of indolicidin, an antibacterial peptide of innate immune origin
Article Snippet: LPS of wild S. typhimurium was obtained from Difco Laboratories. .. The gram-negative bacterial strains S. typhimurium 3261 PNP2 Gro A mutant and E. coli BL21 (αD3) and Streptococcus group A were used for radial diffusion assay.

Article Title: Structure-function analyses involving palindromic analogs of tritrypticin suggest autonomy of anti-endotoxin and antibacterial activities
Article Snippet: Lipopolysaccaride (LPS) of wild S. typhimurium was obtained from Difco Laboratories. .. The Gram-negative bacterial strains S. typhimurium 3261 PNP2 Gro A mutant, E. coli BL21(λD3), P. aeruginosa , and A. tumefaciens were used for radial diffusion assay.

Isolation:

Article Title: Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry
Article Snippet: The E. coli O157:H7 (E8) and S. Typhimurium (S9) strains were isolated from chickens in our laboratory. .. The strains of E. coli O157:H7 and S. Typhimurium were kept at -80°C in Luria-Bertani broth (LB, Difco) containing 20% (v/v) glycerol for long-term preservation and maintained on LB slants at 4°C for temporary use.

Article Title: Changes in the Microbiological Characteristics of Korean Native Cattle (Hanwoo) Beef Exposed to Ultraviolet (UV) Irradiation Prior to Refrigeration
Article Snippet: A different agar medium was used for the selective isolation of different pathogens. .. MacConkey’s agar (Difco) medium was used to inoculate the S. Typhimurium, E. coli O157:H7 was inoculated into the MacConkey’s sorbitol agar (Difco) medium.

Preserving:

Article Title: Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry
Article Snippet: .. The strains of E. coli O157:H7 and S. Typhimurium were kept at -80°C in Luria-Bertani broth (LB, Difco) containing 20% (v/v) glycerol for long-term preservation and maintained on LB slants at 4°C for temporary use. .. For experimental preparation, a loop inoculum was transferred to LB broth and incubated for 24 h at 37°C for each food-borne microbial strain.

Diffusion-based Assay:

Article Title: Plasticity in structure and interactions is critical for the action of indolicidin, an antibacterial peptide of innate immune origin
Article Snippet: LPS of wild S. typhimurium was obtained from Difco Laboratories. .. The gram-negative bacterial strains S. typhimurium 3261 PNP2 Gro A mutant and E. coli BL21 (αD3) and Streptococcus group A were used for radial diffusion assay.

Article Title: Structure-function analyses involving palindromic analogs of tritrypticin suggest autonomy of anti-endotoxin and antibacterial activities
Article Snippet: Lipopolysaccaride (LPS) of wild S. typhimurium was obtained from Difco Laboratories. .. The Gram-negative bacterial strains S. typhimurium 3261 PNP2 Gro A mutant, E. coli BL21(λD3), P. aeruginosa , and A. tumefaciens were used for radial diffusion assay.

Immunohistochemistry:

Article Title: Effects of mannoprotein E1 in liquid diet on inflammatory response and TLR5 expression in the gut of rats infected by Salmonella typhimurium
Article Snippet: Colony identification was made its morphological characteristics and agglutination with specific anti-sera to Salmonella poly A and S. typhimurium (Difco). .. Fixed ileum was embedded in paraffin blocks, from which 5-μm serial sections were cut for immunohistochemical studies.

Microscopy:

Article Title: Real-time imaging of type III secretion: Salmonella SipA injection into host cells
Article Snippet: Immunofluorescence Microscopy. .. S. typhimurium was stained with a rabbit anti- Salmonella O -1,4,5,12 ( ) antiserum (Difco) and goat anti-rabbit-FITC , anti-rabbit-7-amino-4-methylcoumarin-3-acetic acid (see ) or anti-rabbit-rhodamine (see Figs. and ) conjugate.

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿
Article Snippet: Paragraph title: Automated microscopy assay for S . Typhimurium binding. ... S. Typhimurium was visualized by indirect immunofluorescence using a rabbit anti- S. Typhimurium antibody (Difco) and a fluorescein-5-isothiocyanate (FITC)-labeled goat anti-rabbit secondary antibody (Jackson).

Sampling:

Article Title: Effects of mannoprotein E1 in liquid diet on inflammatory response and TLR5 expression in the gut of rats infected by Salmonella typhimurium
Article Snippet: Paragraph title: Tissue Sampling ... Colony identification was made its morphological characteristics and agglutination with specific anti-sera to Salmonella poly A and S. typhimurium (Difco).

Incubation:

Article Title: Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)
Article Snippet: .. A single colony of each strain of E. coli O157:H7, S. Typhimurium, and L. monocytogenes cultured from stocks on tryptic soy agar (TSA) (Difco, Becton, Dickinson, Sparks, MD), were inoculated individually into 15 ml tryptic soy broth (TSB) (Difco), incubated at 37°C for 24 h, and then collected by centrifugation at 4,000 × g for 20 min at 4°C. .. After three washes with 0.2% peptone water (PW) (Bacto, Becton, Dickinson, Sparks, MD), suspended pellets of the three pathogens were combined to constitute a mixed-pathogen-species culture cocktail corresponding to approximately 108 to 109 CFU/ml.

Article Title: Real-time imaging of type III secretion: Salmonella SipA injection into host cells
Article Snippet: For immunostaining of effector proteins in the bacterial cytosol (see Figs. and ), S. typhimurium immobilized on gelatin-coated cover slips (no host cell contact) or infection assays were fixed for 20 min at 22°C in 4% paraformaldehyde/4% sucrose/PBS, incubated in 20% sucrose in PBS for 10 min at 22°C, permeabilized with buffer A (50 mM EDTA/20 mM Tris·HCl/1.8 g/liter glucose/0.1% Triton X-100, pH 8) for 5 min at 22°C, washed three times in buffer B (10 mM EDTA/25 mM Tris·HCl/1.8 g/liter glucose, pH 8) at 22°C, and incubated for 1 h in buffer B plus 5 g/liter lysozyme at 4°C. .. S. typhimurium was stained with a rabbit anti- Salmonella O -1,4,5,12 ( ) antiserum (Difco) and goat anti-rabbit-FITC , anti-rabbit-7-amino-4-methylcoumarin-3-acetic acid (see ) or anti-rabbit-rhodamine (see Figs. and ) conjugate.

Article Title: Evaluation of Sanitizing Methods for Reducing Microbial Contamination on Fresh Strawberry, Cherry Tomato, and Red Bayberry
Article Snippet: The strains of E. coli O157:H7 and S. Typhimurium were kept at -80°C in Luria-Bertani broth (LB, Difco) containing 20% (v/v) glycerol for long-term preservation and maintained on LB slants at 4°C for temporary use. .. For experimental preparation, a loop inoculum was transferred to LB broth and incubated for 24 h at 37°C for each food-borne microbial strain.

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿
Article Snippet: Fixation was performed with 4% paraformaldehyde, 4% sucrose in phosphate buffer saline (PBS), followed by incubation in 20% sucrose in PBS overnight. .. S. Typhimurium was visualized by indirect immunofluorescence using a rabbit anti- S. Typhimurium antibody (Difco) and a fluorescein-5-isothiocyanate (FITC)-labeled goat anti-rabbit secondary antibody (Jackson).

Article Title: Changes in the Microbiological Characteristics of Korean Native Cattle (Hanwoo) Beef Exposed to Ultraviolet (UV) Irradiation Prior to Refrigeration
Article Snippet: MacConkey’s agar (Difco) medium was used to inoculate the S. Typhimurium, E. coli O157:H7 was inoculated into the MacConkey’s sorbitol agar (Difco) medium. .. The inhibitory effect was measured by bacterial colony counting after a 24 h incubation at 37℃.

Transfection:

Article Title: Real-time imaging of type III secretion: Salmonella SipA injection into host cells
Article Snippet: Cos7 cells were seeded onto glass coverslips in DMEM plus 5% FCS (PAA, Linz, Austria), transfected with PolyFect (Qiagen, Valencia, CA), and grown to 70-80% confluency. .. S. typhimurium was stained with a rabbit anti- Salmonella O -1,4,5,12 ( ) antiserum (Difco) and goat anti-rabbit-FITC , anti-rabbit-7-amino-4-methylcoumarin-3-acetic acid (see ) or anti-rabbit-rhodamine (see Figs. and ) conjugate.

Cell Culture:

Article Title: Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.)
Article Snippet: .. A single colony of each strain of E. coli O157:H7, S. Typhimurium, and L. monocytogenes cultured from stocks on tryptic soy agar (TSA) (Difco, Becton, Dickinson, Sparks, MD), were inoculated individually into 15 ml tryptic soy broth (TSB) (Difco), incubated at 37°C for 24 h, and then collected by centrifugation at 4,000 × g for 20 min at 4°C. .. After three washes with 0.2% peptone water (PW) (Bacto, Becton, Dickinson, Sparks, MD), suspended pellets of the three pathogens were combined to constitute a mixed-pathogen-species culture cocktail corresponding to approximately 108 to 109 CFU/ml.

Article Title: Evaluation of Near-Infrared Pasteurization in Controlling Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes in Ready-To-Eat Sliced Ham
Article Snippet: .. Each strain of S. Typhimurium, E. coli O157:H7, and L. monocytogenes was cultured in 5 ml of TSB at 37°C for 24 h, followed by centrifugation (4,000 × g for 20 min at 4°C) and washing three times with buffered peptone water (BPW; Difco, Sparks, MD). ..

Article Title: Commensal Akkermansia muciniphila Exacerbates Gut Inflammation in Salmonella Typhimurium-Infected Gnotobiotic Mice
Article Snippet: .. The SIHUMI members and S. Typhimurium were cultured in yeast casitone fatty acid (YCFA) medium while A. muciniphila was cultured in Columbia broth (Difco). .. All strains were cultured under strictly anoxic conditions using N2: CO2 (80∶20; v∶v) as the gas phase.

Staining:

Article Title: Real-time imaging of type III secretion: Salmonella SipA injection into host cells
Article Snippet: .. S. typhimurium was stained with a rabbit anti- Salmonella O -1,4,5,12 ( ) antiserum (Difco) and goat anti-rabbit-FITC , anti-rabbit-7-amino-4-methylcoumarin-3-acetic acid (see ) or anti-rabbit-rhodamine (see Figs. and ) conjugate. .. SipAM45 and/or SopEHA were stained with monoclonal mouse anti-M45 [kindly provided by P. Hearing (State University of New York, Stony Brook)] or rabbit anti-HA (Santa Cruz Biotechnology) and goat anti-mouse-Cy3 (see Figs. , , and ) or anti-mouse-Cy5 (see Figs. and ) and/or goat α-rabbit-FITC (see ) conjugates (Jackson ImmunoResearch).

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿
Article Snippet: S. Typhimurium was visualized by indirect immunofluorescence using a rabbit anti- S. Typhimurium antibody (Difco) and a fluorescein-5-isothiocyanate (FITC)-labeled goat anti-rabbit secondary antibody (Jackson). .. Nuclei were stained with DAPI (4′,6′-diamidino-2-phenylindole) after a 5-min permeabilization step using 0.1% Triton X-100.

Modification:

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿
Article Snippet: HeLa cells were seeded 24 h prior to infection in 96-well μ-Clear plates (half-size; Greiner) at 6,000 cells per well in Dulbecco modified Eagle medium (DMEM), 10% fetal calf serum (FCS). .. S. Typhimurium was visualized by indirect immunofluorescence using a rabbit anti- S. Typhimurium antibody (Difco) and a fluorescein-5-isothiocyanate (FITC)-labeled goat anti-rabbit secondary antibody (Jackson).

Western Blot:

Article Title: Effects of mannoprotein E1 in liquid diet on inflammatory response and TLR5 expression in the gut of rats infected by Salmonella typhimurium
Article Snippet: Colony identification was made its morphological characteristics and agglutination with specific anti-sera to Salmonella poly A and S. typhimurium (Difco). .. Samples from the ileum, jejunum and colon were taken and placed in 10% neutral buffered formalin during 24 hr for Q-PCR and Western-blot assay.

Injection:

Article Title: The role of lipopolysaccharide injected systemically in the reactivation of collagen-induced arthritis in mice
Article Snippet: Varying doses of LPS were dissolved in 100 μl of sterile, pyrogen-free saline and injected i.p. on day 50. .. In some experiments, LPS from S. enteritidis, S. typhimurium (Difco), and K. neumoniae (Sigma Chemical Co., St. Louis, MO, U.S.A.) and lipid A from E. coli K12D31m4 (Funakoshi Co., Tokyo, Japan) were also i.p. administered.

Binding Assay:

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿
Article Snippet: Paragraph title: Automated microscopy assay for S . Typhimurium binding. ... S. Typhimurium was visualized by indirect immunofluorescence using a rabbit anti- S. Typhimurium antibody (Difco) and a fluorescein-5-isothiocyanate (FITC)-labeled goat anti-rabbit secondary antibody (Jackson).

Immunofluorescence:

Article Title: Real-time imaging of type III secretion: Salmonella SipA injection into host cells
Article Snippet: Immunofluorescence Microscopy. .. S. typhimurium was stained with a rabbit anti- Salmonella O -1,4,5,12 ( ) antiserum (Difco) and goat anti-rabbit-FITC , anti-rabbit-7-amino-4-methylcoumarin-3-acetic acid (see ) or anti-rabbit-rhodamine (see Figs. and ) conjugate.

Article Title: Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking ▿
Article Snippet: .. S. Typhimurium was visualized by indirect immunofluorescence using a rabbit anti- S. Typhimurium antibody (Difco) and a fluorescein-5-isothiocyanate (FITC)-labeled goat anti-rabbit secondary antibody (Jackson). .. Nuclei were stained with DAPI (4′,6′-diamidino-2-phenylindole) after a 5-min permeabilization step using 0.1% Triton X-100.

Generated:

Article Title: Benzyl Isothiocyanate, a Major Component from the Roots of Salvadora Persica Is Highly Active against Gram-Negative Bacteria
Article Snippet: H. influenzae was propagated over-night on GC-agar (Acumedia) supplemented with haemoglobin (1%) (BBL™) and isovitalex enrichment (1%) (BBL™) in 5% CO2 atmosphere generated with gas-packs. .. E. coli MC4100, E. coli D21, P. aeruginosa , and S. typhimurium were propagated overnight in air on Luria Agar Base, Miller (Difco™).

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    Difco s typhimurium
    MET-1 pre-treatment does not prevent S. <t>typhimurium</t> intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.
    S Typhimurium, supplied by Difco, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s typhimurium/product/Difco
    Average 90 stars, based on 2 article reviews
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    s typhimurium - by Bioz Stars, 2020-01
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    77
    Difco s enterica serovar typhimurium dt104 strain 11601
    Comparison of nonculturable S. <t>enterica</t> <t>serovar</t> <t>Typhimurium</t> <t>DT104</t> 11601 (∼5 × 10 4 CFU/ml) exposed to an 80°C (56°C internal temperature) upshift for 15 s (left) versus no temperature upshift (right) followed by inoculation on TSA at 37°C for 24 h. Selected colonies from the left plate were tested with Gram stain, TSI agar slants, and API 20E and were confirmed to be Salmonella .
    S Enterica Serovar Typhimurium Dt104 Strain 11601, supplied by Difco, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s enterica serovar typhimurium dt104 strain 11601/product/Difco
    Average 77 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    s enterica serovar typhimurium dt104 strain 11601 - by Bioz Stars, 2020-01
    77/100 stars
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    MET-1 pre-treatment does not prevent S. typhimurium intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: MET-1 pre-treatment does not prevent S. typhimurium intracellular invasion of Caco-2 epithelial cells. Live (or heat-killed) S. typhimurium were added to Caco-2 cell monolayers at an MOI of 100:1 and incubated for 1 hour. Extracellular bacteria were then killed by the addition of gentamicin (100 μg/mL). One hour later, Caco-2 cells were lysed and intracellular bacteria were enumerated using serial dilutions plated on MacConkey agar plates containing 100 μg/mL streptomycin. There were no significant differences between S. typhimurium -infected cells pretreated with MET-1 (MET-1 + Sal) or saline (Saline + Sal) (p > 0.05). No bacterial growth was seen in cell monolayers treated with saline only (Saline), MET-1 only (MET-1), heat-killed S. typhimurium only (Saline + HK Sal) or MET-1 pretreated cell monolayers treated with heat-killed S. typhimurium (MET-1 + HK Sal). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 3, (p > 0.05) NS = not significant.

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: Incubation, Infection

    MET-1 pretreatment prevented the disruption of ZO-1 and claudin-1 cellular localization in the cecum caused by S. typhimurium . ( a ) ZO-1 immunofluorescence staining (green) in ceca fixed in 10% formalin (400× magnification). Nuclei were stained using Hoechst (blue). White arrows indicate green staining of ZO-1 protein at the membrane of intestinal epithelial cells. VC = uninfected mice pretreated with vehicle control; MET-1 = uninfected mice pretreated with MET-1; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1. VC + S . Tm. mice had a loss of ZO-1 membrane staining that was attenuated in MET-1 + S . Tm. mice. ( b ) Expression of ZO-1 measured by Western blot analysis (S, Triton X-soluble fraction; I, Triton X-insoluble fraction, sample blot shown in top right of panel). Consistent with data in panel ( a ), densitometric values calculated for the ratio of ZO-1 to β-actin showed that MET-1 + S . Tm. mice had increased expression of ZO-1 in insoluble fractions compared to VC + S . Tm. mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction (*p

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: MET-1 pretreatment prevented the disruption of ZO-1 and claudin-1 cellular localization in the cecum caused by S. typhimurium . ( a ) ZO-1 immunofluorescence staining (green) in ceca fixed in 10% formalin (400× magnification). Nuclei were stained using Hoechst (blue). White arrows indicate green staining of ZO-1 protein at the membrane of intestinal epithelial cells. VC = uninfected mice pretreated with vehicle control; MET-1 = uninfected mice pretreated with MET-1; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1. VC + S . Tm. mice had a loss of ZO-1 membrane staining that was attenuated in MET-1 + S . Tm. mice. ( b ) Expression of ZO-1 measured by Western blot analysis (S, Triton X-soluble fraction; I, Triton X-insoluble fraction, sample blot shown in top right of panel). Consistent with data in panel ( a ), densitometric values calculated for the ratio of ZO-1 to β-actin showed that MET-1 + S . Tm. mice had increased expression of ZO-1 in insoluble fractions compared to VC + S . Tm. mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction (*p

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: Immunofluorescence, Staining, Mouse Assay, Infection, Expressing, Western Blot

    MET-1 inhibited S. typhimurium in vitro but did not inhibit its growth in vivo . ( a ) MET-1 (1:2, 1:100 and 1:300 dilutions) was incubated with S. typhimurium ( S . Tm.), aliquots were withdrawn at times indicated, plated, counted, and plotted as log 10 CFU. MET-1 inhibited the growth of S. Tm. relative to the S . Tm. control, although MET-1 was unable to completely kill S. Tm. even at a high (1:2) concentration. ( b ) Colons were harvested 48 hours after S. Tm. infection, homogenized in sterile PBS, and plated. Infected mice pretreated with MET-1 (MET-1 + S . Tm.) showed no significant reduction of S. Tm. counts in the colon (p > 0.05) compared to mice pretreated with vehicle control (VC + S . Tm.). S. Tm. was not detected in the colon of uninfected mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction. VC = uninfected mice pretreated with vehicle control, n = 18; MET-1 = uninfected mice pretreated with MET-1, n = 18; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 22; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 21.

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: MET-1 inhibited S. typhimurium in vitro but did not inhibit its growth in vivo . ( a ) MET-1 (1:2, 1:100 and 1:300 dilutions) was incubated with S. typhimurium ( S . Tm.), aliquots were withdrawn at times indicated, plated, counted, and plotted as log 10 CFU. MET-1 inhibited the growth of S. Tm. relative to the S . Tm. control, although MET-1 was unable to completely kill S. Tm. even at a high (1:2) concentration. ( b ) Colons were harvested 48 hours after S. Tm. infection, homogenized in sterile PBS, and plated. Infected mice pretreated with MET-1 (MET-1 + S . Tm.) showed no significant reduction of S. Tm. counts in the colon (p > 0.05) compared to mice pretreated with vehicle control (VC + S . Tm.). S. Tm. was not detected in the colon of uninfected mice. Data were analyzed using a 1-way ANOVA with Bonferroni correction. VC = uninfected mice pretreated with vehicle control, n = 18; MET-1 = uninfected mice pretreated with MET-1, n = 18; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 22; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 21.

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: In Vitro, In Vivo, Incubation, Concentration Assay, Infection, Mouse Assay

    Pro-inflammatory cytokine levels were reduced in S. typhimurium -infected mice pretreated with MET-1. ( a ) Serum cytokine levels were measured 48 hours post-infection using a Bio-Plex Pro mouse cytokine magnetic bead kit. Of the cytokines measured, TNF-α, IL-1β, IL-12p40, IL-12p70, GM-CSF, eotaxin, MIP-1β, and MCP-1 were all significantly reduced in infected mice pretreated with MET-1 (MET-1 + S . Tm.) compared to infected mice pretreated with vehicle control (VC + S . Tm.). There were no significant differences in cytokine concentrations between uninfected mice pretreated with MET-1 or vehicle control (data not shown). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 7 for each group (*p

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: Pro-inflammatory cytokine levels were reduced in S. typhimurium -infected mice pretreated with MET-1. ( a ) Serum cytokine levels were measured 48 hours post-infection using a Bio-Plex Pro mouse cytokine magnetic bead kit. Of the cytokines measured, TNF-α, IL-1β, IL-12p40, IL-12p70, GM-CSF, eotaxin, MIP-1β, and MCP-1 were all significantly reduced in infected mice pretreated with MET-1 (MET-1 + S . Tm.) compared to infected mice pretreated with vehicle control (VC + S . Tm.). There were no significant differences in cytokine concentrations between uninfected mice pretreated with MET-1 or vehicle control (data not shown). Data were analyzed using a 1-way ANOVA with Bonferroni correction, n = 7 for each group (*p

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: Infection, Mouse Assay

    Translocation of S. typhimurium to the spleen was reduced in MET-1 pretreated mice. Spleens were harvested 48 hours after S. typhimurium infection, weighed, and homogenized in 1 mL PBS. Serial dilutions were plated on MacConkey agar plates containing 100 μg/mL streptomycin. Plates were incubated at 37 °C for 24 hrs, and the resulting colonies were counted. Infected mice pretreated with MET-1 (MET-1 + S . Tm., n = 13) had reduced S. typhimurium counts in the spleen (*p

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: Translocation of S. typhimurium to the spleen was reduced in MET-1 pretreated mice. Spleens were harvested 48 hours after S. typhimurium infection, weighed, and homogenized in 1 mL PBS. Serial dilutions were plated on MacConkey agar plates containing 100 μg/mL streptomycin. Plates were incubated at 37 °C for 24 hrs, and the resulting colonies were counted. Infected mice pretreated with MET-1 (MET-1 + S . Tm., n = 13) had reduced S. typhimurium counts in the spleen (*p

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: Translocation Assay, Mouse Assay, Infection, Incubation

    MET-1 attenuates local neutrophil infiltration induced by S. typhimurium infection in cecum (a) Representative images of myeloperoxidase (MPO) immunofluorescence staining for neutrophils (green) in ceca. The area in the box (left) is displayed at a higher magnification in the panels on the right. Nucleic acid was stained using Hoechst (blue). S. typhimurium -infected mice pretreated with vehicle control (VC + S . Tm.) had more abundant immunostaining in the ceca compared to uninfected controls (VC), and was significantly reduced in the ceca of infected mice pretreated with MET-1 (MET-1 + S . Tm. mice). No MPO staining was noted in secondary antibody control slides for each group (not shown). ( b ) Quantification of MPO immunostaining, shown by the ratio of MPO (green immunofluorescence) staining to number of cells (blue nuclei). Data analyzed using 1-way ANOVA with Bonferroni correction, ***p

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: MET-1 attenuates local neutrophil infiltration induced by S. typhimurium infection in cecum (a) Representative images of myeloperoxidase (MPO) immunofluorescence staining for neutrophils (green) in ceca. The area in the box (left) is displayed at a higher magnification in the panels on the right. Nucleic acid was stained using Hoechst (blue). S. typhimurium -infected mice pretreated with vehicle control (VC + S . Tm.) had more abundant immunostaining in the ceca compared to uninfected controls (VC), and was significantly reduced in the ceca of infected mice pretreated with MET-1 (MET-1 + S . Tm. mice). No MPO staining was noted in secondary antibody control slides for each group (not shown). ( b ) Quantification of MPO immunostaining, shown by the ratio of MPO (green immunofluorescence) staining to number of cells (blue nuclei). Data analyzed using 1-way ANOVA with Bonferroni correction, ***p

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: Infection, Immunofluorescence, Staining, Mouse Assay, Immunostaining

    MET-1 attenuated systemic markers of disease in S. typhimurium -infected mice. ( a ) MET-1 attenuated weight loss in S. typhimurium infected mice. Following oral infection with S. typhimurium , mice were weighed daily and the percent change in weight from 0 to 48 hours is shown. VC = uninfected mice pretreated with vehicle control, n = 16; MET-1 = uninfected mice pretreated with MET-1, n = 16; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 18; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 18. Data were analyzed with a 2-way ANOVA using Bonferroni correction (*p

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: MET-1 attenuated systemic markers of disease in S. typhimurium -infected mice. ( a ) MET-1 attenuated weight loss in S. typhimurium infected mice. Following oral infection with S. typhimurium , mice were weighed daily and the percent change in weight from 0 to 48 hours is shown. VC = uninfected mice pretreated with vehicle control, n = 16; MET-1 = uninfected mice pretreated with MET-1, n = 16; VC + S . Tm. = S. typhimurium -infected mice pretreated with vehicle control, n = 18; MET-1 + S . Tm. = S. typhimurium -infected mice pretreated with MET-1, n = 18. Data were analyzed with a 2-way ANOVA using Bonferroni correction (*p

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: Infection, Mouse Assay

    MET-1 did not enhance intestinal mucin production in ceca of S. typhimurium - infected mice. ( a ) Alcian blue staining (blue) of ceca fixed in 10% formalin were counterstained with nuclear red fast solution (red). ( b ) Goblet cells were enumerated from 10 random high-powered field views (400× magnification) spanning from muscularis mucosa to surface epithelium on Alcian blue stained sections. Infected mice pretreated with vehicle control (VC + S . Tm.) had less mucin staining than uninfected mice pretreated with vehicle control (VC + S . Tm.). However, there was no statistically significant difference between infected mice pretreated with MET-1 (MET-1 + S . Tm.) and vehicle control (VC + S . Tm.). ( c ) MUC2 immunofluorescence staining (green) in ceca fixed in Methanol-Carnoy’s solution. Nucleic acid was stained using DAPI (blue). Uninfected mice pretreated with VC or MET-1 had similar localization of MUC2 staining that differed from VC + S . Tm. mice. Again, MUC2 staining in MET-1 + S . Tm. mice more closely resembled VC + S . Tm. mice than uninfected controls. Quantification shown in (d). Data were analyzed using a 1-way ANOVA with Bonferroni correction. n = 3, (**p

    Journal: Scientific Reports

    Article Title: Administration of defined microbiota is protective in a murine Salmonella infection model

    doi: 10.1038/srep16094

    Figure Lengend Snippet: MET-1 did not enhance intestinal mucin production in ceca of S. typhimurium - infected mice. ( a ) Alcian blue staining (blue) of ceca fixed in 10% formalin were counterstained with nuclear red fast solution (red). ( b ) Goblet cells were enumerated from 10 random high-powered field views (400× magnification) spanning from muscularis mucosa to surface epithelium on Alcian blue stained sections. Infected mice pretreated with vehicle control (VC + S . Tm.) had less mucin staining than uninfected mice pretreated with vehicle control (VC + S . Tm.). However, there was no statistically significant difference between infected mice pretreated with MET-1 (MET-1 + S . Tm.) and vehicle control (VC + S . Tm.). ( c ) MUC2 immunofluorescence staining (green) in ceca fixed in Methanol-Carnoy’s solution. Nucleic acid was stained using DAPI (blue). Uninfected mice pretreated with VC or MET-1 had similar localization of MUC2 staining that differed from VC + S . Tm. mice. Again, MUC2 staining in MET-1 + S . Tm. mice more closely resembled VC + S . Tm. mice than uninfected controls. Quantification shown in (d). Data were analyzed using a 1-way ANOVA with Bonferroni correction. n = 3, (**p

    Article Snippet: CFU/mL of S. typhimurium were calculated by serial dilutions plated onto MacConkey agar (Difco Laboratories, Becton, Dickinson and Company, MD, USA) supplemented with streptomycin (100 μg/mL).

    Techniques: Infection, Mouse Assay, Staining, Immunofluorescence

    Virulence of Δ zntA Δ zitB mutant S. Typhimurium is attenuated in NO·-producing mice. Solid lines represent the median competitive index (CI) for each organ. The dotted line represents the expected CI if neither strain has a competitive advantage. (A) In wild-type C3H/HeOuJ mice, a Δ zntA Δ zitB mutant has a significant competitive disadvantage compared to wild type ( P = 0.002 by Wilcoxon signed-rank test to a hypothetical median of 1 for both spleen and liver). (B) In C3H/HeOuJ mice that cannot produce NO· due to treatment with 500 µg ml −1 l - N 6 -(1-iminoethyl)lysine dihydrochloride ( l -NIL), the mutant no longer has a statistically significant disadvantage compared to wild type, and the CIs are significantly different (*, P = 0.007 for spleen and P

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: Virulence of Δ zntA Δ zitB mutant S. Typhimurium is attenuated in NO·-producing mice. Solid lines represent the median competitive index (CI) for each organ. The dotted line represents the expected CI if neither strain has a competitive advantage. (A) In wild-type C3H/HeOuJ mice, a Δ zntA Δ zitB mutant has a significant competitive disadvantage compared to wild type ( P = 0.002 by Wilcoxon signed-rank test to a hypothetical median of 1 for both spleen and liver). (B) In C3H/HeOuJ mice that cannot produce NO· due to treatment with 500 µg ml −1 l - N 6 -(1-iminoethyl)lysine dihydrochloride ( l -NIL), the mutant no longer has a statistically significant disadvantage compared to wild type, and the CIs are significantly different (*, P = 0.007 for spleen and P

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: Mutagenesis, Mouse Assay

    Expression of zinc transport systems in S. Typhimurium. qPCR data are presented as a positive fold change of treated compared to untreated cells with zinc efflux systems shown in blue and zinc acquisition systems shown in red. The solid line indicates a fold change of 1 to delineate between upregulation ( > 1) and downregulation (

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: Expression of zinc transport systems in S. Typhimurium. qPCR data are presented as a positive fold change of treated compared to untreated cells with zinc efflux systems shown in blue and zinc acquisition systems shown in red. The solid line indicates a fold change of 1 to delineate between upregulation ( > 1) and downregulation (

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    ICP-MS analysis of total cellular zinc content in S. Typhimurium following NO· treatment and transcriptional monitoring of NO· sensed by cells. (A) S. Typhimurium cells at an OD 600 of ≈1 were treated with 2 mM diethylamine NONOate (DEANO), and total cellular zinc was measured at various times posttreatment. By 5 min posttreatment, total cellular zinc had fallen significantly compared to untreated cells, suggesting that zinc is effluxed from the cell following NO· treatment. Zinc levels gradually recovered to baseline levels over the course of 60 min. Statistical significance was determined by unpaired two-tailed t test; * indicates P values of

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: ICP-MS analysis of total cellular zinc content in S. Typhimurium following NO· treatment and transcriptional monitoring of NO· sensed by cells. (A) S. Typhimurium cells at an OD 600 of ≈1 were treated with 2 mM diethylamine NONOate (DEANO), and total cellular zinc was measured at various times posttreatment. By 5 min posttreatment, total cellular zinc had fallen significantly compared to untreated cells, suggesting that zinc is effluxed from the cell following NO· treatment. Zinc levels gradually recovered to baseline levels over the course of 60 min. Statistical significance was determined by unpaired two-tailed t test; * indicates P values of

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: Mass Spectrometry, Two Tailed Test

    Free intracellular zinc levels increase in a Δ zntA Δ zitB S. Typhimurium mutant during macrophage infection in response to NO· production. (A) Changes in NO· production are shown at the time of infection (0 h) and 13 h postinfection in the presence or absence of the NOS inhibitor l -NMMA. IFN-γ-primed murine macrophages infected with either wild-type (blue) or Δ zntA Δ zitB (green) S. Typhimurium produced significant levels of NO· after 13 h in the absence of the NOS inhibitor l -NMMA ( P

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: Free intracellular zinc levels increase in a Δ zntA Δ zitB S. Typhimurium mutant during macrophage infection in response to NO· production. (A) Changes in NO· production are shown at the time of infection (0 h) and 13 h postinfection in the presence or absence of the NOS inhibitor l -NMMA. IFN-γ-primed murine macrophages infected with either wild-type (blue) or Δ zntA Δ zitB (green) S. Typhimurium produced significant levels of NO· after 13 h in the absence of the NOS inhibitor l -NMMA ( P

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: Mutagenesis, Infection, Produced

    ZntA and ZitB are the primary zinc efflux transporters in S. Typhimurium. (A) A Δ zntA mutant (pink) was impaired for growth, represented by a delayed exit from lag phase, in both 0.125 mM ZnSO 4 and 0.25 mM ZnSO 4 , while a Δ zntB mutant (aqua) and a Δ zitB mutant (light green) exhibited growth comparable to wild type (blue) under these conditions. (B) Significance of the growth defect for the Δ zntA mutant in panel A was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. (C) A Δ zntB Δ zitB double mutant (purple) exhibited growth comparable to wild type (blue) when exposed to elevated zinc concentrations. A Δ zntA Δ zntB mutant (red) behaved similarly to a Δ zntA mutant, whereas a Δ zntA Δ zitB mutant (green) displayed a more severe growth delay at 0.125 mM ZnSO 4 and was unable to grow at 0.25 mM ZnSO 4 . A Δ zntA Δ zntB Δ zitB triple mutant (orange) exhibited growth characteristics identical to those of a Δ zntA Δ zitB double mutant. (D) Significance of the growth defects for the mutants in panel C was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. Statistical significance of the growth defects was determined by unpaired two-tailed t test, and an asterisk indicates P values of

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: ZntA and ZitB are the primary zinc efflux transporters in S. Typhimurium. (A) A Δ zntA mutant (pink) was impaired for growth, represented by a delayed exit from lag phase, in both 0.125 mM ZnSO 4 and 0.25 mM ZnSO 4 , while a Δ zntB mutant (aqua) and a Δ zitB mutant (light green) exhibited growth comparable to wild type (blue) under these conditions. (B) Significance of the growth defect for the Δ zntA mutant in panel A was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. (C) A Δ zntB Δ zitB double mutant (purple) exhibited growth comparable to wild type (blue) when exposed to elevated zinc concentrations. A Δ zntA Δ zntB mutant (red) behaved similarly to a Δ zntA mutant, whereas a Δ zntA Δ zitB mutant (green) displayed a more severe growth delay at 0.125 mM ZnSO 4 and was unable to grow at 0.25 mM ZnSO 4 . A Δ zntA Δ zntB Δ zitB triple mutant (orange) exhibited growth characteristics identical to those of a Δ zntA Δ zitB double mutant. (D) Significance of the growth defects for the mutants in panel C was determined by calculating the mean time required to reach 50% of the maximum final OD 600 for each strain. Statistical significance of the growth defects was determined by unpaired two-tailed t test, and an asterisk indicates P values of

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: Mutagenesis, Two Tailed Test

    A model of zinc homeostasis in Salmonella Typhimurium. Under conditions of zinc deficiency, zinc is not available to bind to the Zur repressor, leading to expression of the ZnuABC zinc acquisition system. ZupT, whose regulation is uncharacterized, has also been shown to contribute to zinc acquisition. When zinc is abundant, Zur bound to zinc represses ZnuABC expression. In addition, free cytoplasmic zinc binds to the transcriptional activator ZntR to induce expression of the ZntA zinc efflux system. Zinc efflux in S. Typhimurium is also mediated by ZitB. Under conditions of nitrosative stress, S -nitrosylation of cysteine ligands in zinc metalloproteins leads to mobilization of free intracellular zinc. The zinc efflux activities of ZntA and ZitB are required for the resistance of S. Typhimurium to nitrosative stress.

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: A model of zinc homeostasis in Salmonella Typhimurium. Under conditions of zinc deficiency, zinc is not available to bind to the Zur repressor, leading to expression of the ZnuABC zinc acquisition system. ZupT, whose regulation is uncharacterized, has also been shown to contribute to zinc acquisition. When zinc is abundant, Zur bound to zinc represses ZnuABC expression. In addition, free cytoplasmic zinc binds to the transcriptional activator ZntR to induce expression of the ZntA zinc efflux system. Zinc efflux in S. Typhimurium is also mediated by ZitB. Under conditions of nitrosative stress, S -nitrosylation of cysteine ligands in zinc metalloproteins leads to mobilization of free intracellular zinc. The zinc efflux activities of ZntA and ZitB are required for the resistance of S. Typhimurium to nitrosative stress.

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: Expressing

    Zinc efflux by ZntA and ZitB is required for S. Typhimurium resistance to nitrosative stress in vitro . (A) Single zinc efflux mutants (pink, aqua, and light green) and a Δ zntA Δ zntB double mutant (red) were no more sensitive to NO· generated by the donor spermine NONOate (SperNO) than wild-type cells (blue). However, a Δ zntA Δ zitB double mutant (green) exhibited significantly delayed growth, indicating enhanced NO· sensitivity (*, P

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: Zinc efflux by ZntA and ZitB is required for S. Typhimurium resistance to nitrosative stress in vitro . (A) Single zinc efflux mutants (pink, aqua, and light green) and a Δ zntA Δ zntB double mutant (red) were no more sensitive to NO· generated by the donor spermine NONOate (SperNO) than wild-type cells (blue). However, a Δ zntA Δ zitB double mutant (green) exhibited significantly delayed growth, indicating enhanced NO· sensitivity (*, P

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: In Vitro, Mutagenesis, Generated

    Classification of proteins in the S. Typhimurium S -nitrosoproteome. (A) Functional classification of proteins with cysteine residues modified by NO· treatment. A total of 141 modified proteins were identified, and classification is shown as a percentage of the total. (B) Of the proteins modified by NO·, 15 (~10%) were found to be zinc metalloproteins. The metalloproteins are sorted by functional category and listed in numerical order by gene identifier in S. Typhimurium strain 14028s.

    Journal: mBio

    Article Title: Nitric Oxide Disrupts Zinc Homeostasis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/mBio.01040-18

    Figure Lengend Snippet: Classification of proteins in the S. Typhimurium S -nitrosoproteome. (A) Functional classification of proteins with cysteine residues modified by NO· treatment. A total of 141 modified proteins were identified, and classification is shown as a percentage of the total. (B) Of the proteins modified by NO·, 15 (~10%) were found to be zinc metalloproteins. The metalloproteins are sorted by functional category and listed in numerical order by gene identifier in S. Typhimurium strain 14028s.

    Article Snippet: S. Typhimurium was grown aerobically in Luria-Bertani (L B; Difco) medium at 37°C with shaking at 250 rpm.

    Techniques: Functional Assay, Modification

    Comparison of nonculturable S. enterica serovar Typhimurium DT104 11601 (∼5 × 10 4 CFU/ml) exposed to an 80°C (56°C internal temperature) upshift for 15 s (left) versus no temperature upshift (right) followed by inoculation on TSA at 37°C for 24 h. Selected colonies from the left plate were tested with Gram stain, TSI agar slants, and API 20E and were confirmed to be Salmonella .

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Comparison of nonculturable S. enterica serovar Typhimurium DT104 11601 (∼5 × 10 4 CFU/ml) exposed to an 80°C (56°C internal temperature) upshift for 15 s (left) versus no temperature upshift (right) followed by inoculation on TSA at 37°C for 24 h. Selected colonies from the left plate were tested with Gram stain, TSI agar slants, and API 20E and were confirmed to be Salmonella .

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques: Staining

    Comparison of the survival of freshly cultured (gray bars) (∼3 × 10 4 CFU/ml) and nutrient-deficient S. enterica serovar Typhimurium DT104 11601 (∼3 × 10 4 CFU/ml) (black bars) maintained in 7.35 mM BPS microcosms at 21°C for 418 days, after exposure to10 mM H 2 O 2 solution for 100 min. Counts represent the mean of replicate experiments. Points shown on the baseline represent undetectable populations after standard plate count determinations.

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Comparison of the survival of freshly cultured (gray bars) (∼3 × 10 4 CFU/ml) and nutrient-deficient S. enterica serovar Typhimurium DT104 11601 (∼3 × 10 4 CFU/ml) (black bars) maintained in 7.35 mM BPS microcosms at 21°C for 418 days, after exposure to10 mM H 2 O 2 solution for 100 min. Counts represent the mean of replicate experiments. Points shown on the baseline represent undetectable populations after standard plate count determinations.

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques: Cell Culture

    Scanning electron micrographs of S. enterica serovar Typhimurium DT104 11601 exposed to nutrient-limiting conditions for more than 365 days (A) versus cells grown for 24 h in TSB (B). Scale bar, 1 μm.

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Scanning electron micrographs of S. enterica serovar Typhimurium DT104 11601 exposed to nutrient-limiting conditions for more than 365 days (A) versus cells grown for 24 h in TSB (B). Scale bar, 1 μm.

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques:

    Comparison of the recovery of nutrient-deprived S. enterica serovar Typhimurium DT104 11601 previously maintained in microcosms containing 2.3% Instant Ocean at 5°C (○) and 21°C (▴) on catalase-supplemented (broken line) versus nonsupplemented (solid line) TSA plates. Comparable results were obtained with similar microcosms that had been maintained at 37°C for the same period of time. Points shown on the baseline represent undetectable populations after standard plate count determinations.

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Comparison of the recovery of nutrient-deprived S. enterica serovar Typhimurium DT104 11601 previously maintained in microcosms containing 2.3% Instant Ocean at 5°C (○) and 21°C (▴) on catalase-supplemented (broken line) versus nonsupplemented (solid line) TSA plates. Comparable results were obtained with similar microcosms that had been maintained at 37°C for the same period of time. Points shown on the baseline represent undetectable populations after standard plate count determinations.

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques:

    Effect of nutrient limitation on S. enterica serovar Typhimurium DT104 11601 maintained in microcosms at 21°C (full line) containing 7.35 mM (•) and 0.0735 (▪) mM BPS and at 5°C (broken line) in 7.35 (□) and 0.0735 (○) mM BPS over 425 days. Each data point represents a single measurement of the number of CFU per milliliter. Comparable results were obtained with microscosms containing 0.735 mM BPS when the cells were maintained at 5 and 21°C and held for the same period of time (data not shown). Points shown on the baseline represent undetectable populations after standard plate count determinations.

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Effect of nutrient limitation on S. enterica serovar Typhimurium DT104 11601 maintained in microcosms at 21°C (full line) containing 7.35 mM (•) and 0.0735 (▪) mM BPS and at 5°C (broken line) in 7.35 (□) and 0.0735 (○) mM BPS over 425 days. Each data point represents a single measurement of the number of CFU per milliliter. Comparable results were obtained with microscosms containing 0.735 mM BPS when the cells were maintained at 5 and 21°C and held for the same period of time (data not shown). Points shown on the baseline represent undetectable populations after standard plate count determinations.

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques:

    Scanning electron micrograph of representative S. enterica serovar Typhimurium DT104 11601 exhibiting bacillary and coccoid cell morphologies after exposure to nutrient-limiting conditions for more than 365 days. Scale bar, 1 μm.

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Scanning electron micrograph of representative S. enterica serovar Typhimurium DT104 11601 exhibiting bacillary and coccoid cell morphologies after exposure to nutrient-limiting conditions for more than 365 days. Scale bar, 1 μm.

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques:

    Comparison of the survival of a freshly cultured suspension (∼10 4 CFU/ml) (▴) and a nutrient-deficient suspension of S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS microcosms at 21°C (▪) and 5°C (○) for 178 days after exposure to saturated NaCl for 22 to 168 h. Each data point represents a single measurement of the number of CFU per milliliter.

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Comparison of the survival of a freshly cultured suspension (∼10 4 CFU/ml) (▴) and a nutrient-deficient suspension of S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS microcosms at 21°C (▪) and 5°C (○) for 178 days after exposure to saturated NaCl for 22 to 168 h. Each data point represents a single measurement of the number of CFU per milliliter.

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques: Cell Culture

    Resuscitation of nonculturable S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS at 5°C for 273 days when 5.0 ml of a concentration of ∼10 5 CFU/ml was added to 5.0 ml of brain heart infusion broth (final concentration, 5 × 10 4 CFU/ml) and subjected to temperature upshifts to 21°C ( X ), 25°C (▪), and 37°C (•). Cultures maintained at 5°C (♦) without temperature upshift served as controls. Counts represent the mean of replicates. Except for the cultures subjected to a 37°C upshift, the timelines for the other cultures remained on the baseline. The points shown on the baseline represent undetectable populations after standard plate count determinations.

    Journal: Applied and Environmental Microbiology

    Article Title: Induction and Resuscitation of Viable but Nonculturable Salmonella enterica Serovar Typhimurium DT104+

    doi: 10.1128/AEM.69.11.6669-6675.2003

    Figure Lengend Snippet: Resuscitation of nonculturable S. enterica serovar Typhimurium DT104 11601 maintained in 7.35 mM BPS at 5°C for 273 days when 5.0 ml of a concentration of ∼10 5 CFU/ml was added to 5.0 ml of brain heart infusion broth (final concentration, 5 × 10 4 CFU/ml) and subjected to temperature upshifts to 21°C ( X ), 25°C (▪), and 37°C (•). Cultures maintained at 5°C (♦) without temperature upshift served as controls. Counts represent the mean of replicates. Except for the cultures subjected to a 37°C upshift, the timelines for the other cultures remained on the baseline. The points shown on the baseline represent undetectable populations after standard plate count determinations.

    Article Snippet: S. enterica serovar Typhimurium DT104 strain 11601(a kind gift from B. Swaminathan) was cultured on Trypticase soy agar (TSA) (Difco, Detroit, Mich.) and incubated at 37°C.

    Techniques: Concentration Assay