scholarly journals Resistance-Nodulation-Cell Division (RND) Transporter AcrD Confers Resistance to Egg White in Salmonella enterica Serovar Enteritidis

Foods ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 90
Author(s):  
Xiaojie Qin ◽  
Yanhong Liu ◽  
Xianming Shi
Keyword(s):  
Salmonella Enterica ◽  
Egg White ◽  
Luria Bertani ◽  
Morphological Observation ◽  
Wild Type ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Ability Test ◽  
Significant Difference ◽  
Rnd Transporter ◽  
Adhesion And Invasion

The excellent survival ability of Salmonella enterica serovar Enteritidis (S. Enteritidis) in egg white leads to outbreaks of salmonellosis frequently associated with eggs and egg products. Our previous proteomic study showed that the expression of multidrug efflux RND transporter AcrD in S. Enteritidis was significantly up-regulated (4.06-fold) in response to an egg white environment. In this study, the potential role of AcrD in the resistance of S. Enteritidis to egg white was explored by gene deletion, survival ability test, morphological observation, Caco-2 cell adhesion and invasion. It was found that deletion of acrD had no apparent effect on the growth of S. Enteritidis in Luria-Bertani (LB) broth but resulted in a significant (p < 0.05) decrease in resistance of S. Enteritidis to egg white and a small number of cell lysis. Compared to the wild type, a 2-log population reduction was noticed in the ΔacrD mutant with different initial concentrations after incubation with egg white for 3 days. Furthermore, no significant difference (p > 0.05) in the adhesion and invasion was found between the wild type and ΔacrD mutant in LB broth and egg white, but the invasion ability of the ΔacrD mutant in egg white was significantly (p < 0.05) lower than that in LB broth. This indicates that acrD is involved in virulence in Salmonella. Taken together, these results reveal the importance of AcrD on the resistance of S. Enteritidis to egg white.

scholarly journals Virulence and Metabolic Characteristics of Salmonella enterica Serovar Enteritidis Strains with DifferentsefDVariants in Hens

2012 ◽  
Vol 78 (18) ◽  
pp. 6405-6412 ◽  
Author(s):  
Cesar A. Morales ◽  
Jean Guard ◽  
Roxana Sanchez-Ingunza ◽  
Devendra H. Shah ◽  
Mark Harrison
Keyword(s):  
Salmonella Enterica ◽  
Egg Production ◽  
The Body ◽  
Wild Type ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Blood Calcium ◽  
Content Type ◽  
Metabolic Characteristics ◽  
Wide Range ◽  
Metabolic Properties

ABSTRACTSalmonella entericaserovar Enteritidis is one of a fewSalmonella entericaserotypes that has SEF14 fimbriae encoded by thesefoperon, which consists of 4 cotranscribed genes,sefABCD, regulated bysefR. A parental strain was used to construct asefDmutant and its complement, and all 3 strains were compared for gene expression, metabolic properties, and virulence characteristics in hens. Transcription ofsefDby wild type was suppressed at 42°C and absent for the mutant under conditions where the complemented mutant had 103times higher transcription. Growth of the complemented mutant was restricted in comparison to that of the mutant and wild type. Hens infected with the wild type and mutant showed decreased blood calcium and egg production, but infection with the complemented mutant did not. Thus, the absence ofsefDcorrelated with increased metabolic capacity and enhanced virulence of the pathogen. These results suggest that any contribution thatsefDmakes to egg contamination is either unknown or would be limited to early transmission from the environment to the host. Absence ofsefD, either through mutation or by suppression of transcription at the body temperature of the host, may contribute to the virulence ofSalmonella entericaby facilitating growth on a wide range of metabolites.

scholarly journals Salmonella enterica Serovar Enteritidis Pathogenicity Island 1 Is Not Essential for but Facilitates Rapid Systemic Spread in Chickens

2009 ◽  
Vol 77 (7) ◽  
pp. 2866-2875 ◽  
Author(s):  
Taseen S. Desin ◽  
Po-King S. Lam ◽  
Birgit Koch ◽  
Claudia Mickael ◽  
Emil Berberov ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Systemic Infection ◽  
Poultry Meat ◽  
Wild Type ◽  
Secretion Systems ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Systemic Spread ◽  
Mutant Strains ◽  
Vitro Analysis

ABSTRACT Salmonella enterica subsp. enterica serovar Enteritidis is a leading cause of human food-borne illness that is mainly associated with the consumption of contaminated poultry meat and eggs. To cause infection, S. Enteritidis is known to use two type III secretion systems, which are encoded on two salmonella pathogenicity islands, SPI-1 and SPI-2, the first of which is thought to play a major role in invasion and bacterial uptake. In order to study the role of SPI-1 in the colonization of chicken, we constructed deletion mutants affecting the complete SPI-1 region (40 kb) and the invG gene. Both ΔSPI-1 and ΔinvG mutant strains were impaired in the secretion of SipD, a SPI-1 effector protein. In vitro analysis using polarized human intestinal epithelial cells (Caco-2) revealed that both mutant strains were less invasive than the wild-type strain. A similar observation was made when chicken cecal and small intestinal explants were coinfected with the wild-type and ΔSPI-1 mutant strains. Oral challenge of 1-week-old chicken with the wild-type or ΔSPI-1 strains demonstrated that there was no difference in chicken cecal colonization. However, systemic infection of the liver and spleen was delayed in birds that were challenged with the ΔSPI-1 strain. These data demonstrate that SPI-1 facilitates systemic infection but is not essential for invasion and systemic spread of the organism in chickens.

scholarly journals The Global Regulator ArcA Controls Resistance to Reactive Nitrogen and Oxygen Intermediates in Salmonella enterica Serovar Enteritidis

2002 ◽  
Vol 70 (2) ◽  
pp. 451-461 ◽  
Author(s):  
Sangwei Lu ◽  
Patrick B. Killoran ◽  
Ferric C. Fang ◽  
Lee W. Riley
Keyword(s):  
Hydrogen Peroxide ◽  
Salmonella Enterica ◽  
Wide Spectrum ◽  
Clinical Isolates ◽  
Reactive Nitrogen ◽  
Wild Type ◽  
Global Regulator ◽  
Oxygen Intermediates ◽  
Coding Sequence ◽  
Significant Difference

ABSTRACT Salmonella enterica serovar Enteritidis is a major cause of food-borne diseases associated with consumption of shell eggs. Clinical isolates of S. enterica serovar Enteritidis exhibit a wide spectrum of virulence in mice. A highly virulent isolate (SE2472) was previously shown to be more resistant in vitro than other clinical isolates to acidified sodium nitrite (ASN), a generator of reactive nitrogen and oxygen intermediates (RNI/ROI). SE2472 is also more resistant to S-nitrosoglutathione (GSNO) and hydrogen peroxide (H2O2) than an ASN-susceptible isolate of S. enterica serovar Enteritidis (SE8743). To investigate the molecular basis for the RNI/ROI resistance of S. enterica serovar Enteritidis, we transformed a genomic DNA library of SE2472 into SE8743. A plasmid clone conferred upon SE8743 enhanced resistance to ASN, GSNO, and H2O2. The DNA insert in the clone encoded ArcA, a global regulator. An arcA mutant of SE2472 was constructed and was found to be more susceptible to GSNO and hydrogen peroxide but not more susceptible to ASN than wild-type SE2472. The susceptibility of the arcA mutant to GSNO and H2O2 was complemented by a plasmid harboring arcA. The coding sequence of the arcA gene in SE2472 and the coding sequence of the arcA gene in SE8743 were identical, suggesting that the difference in resistance to RNI/ROI maybe due to the activity of genes regulated by ArcA. No significant difference in virulence between the wild type and the arcA mutant of SE2472 was observed in mice. These observations show that arcA is essential for resistance of S. enterica serovar Enteritidis to nitrosative and oxidative stress. However, additional genetic loci may contribute to the resistance to RNI/ROI and unusually high virulence for mice of SE2472.

Cells in shearable and nonshearable regions of Salmonella enterica serovar Enteritidis biofilms are morphologically and physiologically distinct

2009 ◽  
Vol 55 (8) ◽  
pp. 955-966 ◽  
Author(s):  
Anil K. Mangalappalli-Illathu ◽  
John R. Lawrence ◽  
Darren R. Korber
Keyword(s):  
Salmonella Enterica ◽  
Antimicrobial Agents ◽  
Protein Translation ◽  
Physiological Status ◽  
Planktonic Cells ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Significant Difference ◽  
Shock Response ◽  
Enhanced Expression ◽  
In Cells

Cellular morphology, exopolymer chemistry, and protein expression of shearable and nonshearable fractions of Salmonella enterica serovar Enteritidis biofilms were examined. Biofilms were grown at a laminar flow velocity of 0.07 cm·s–1 for ~120 h, resulting in biofilms with a thickness (mean ± SD) of 43 ± 24 µm. An empirically determined shear-inducing flow (1.33 cm·s–1) was then applied for 5 min, effectively reducing biofilm thickness by ~70% and leaving 13 ± 6 µm of nonshearable material and allowing fractionation of biofilm material into shearable and nonshearable regions. In situ lectin binding analyses revealed that there was no significant difference in the exopolymer glycoconjugate composition of the shearable and nonshearable biofilm zones. Length to width indices of cells from nonshearable and shearable biofilm regions as well as planktonic cells from biofilm effluent and continuous culture were determined to be 3.2, 2.3, 2.2, and 1.7, respectively, indicating that the cells in the shearable fraction were morphologically more similar to planktonic cells than the cells in the nonshearable biofilm fraction. Enhanced expression of proteins involved in cold shock response, adaptation, and broad regulatory functions (CspA, GrcA, and Hns, respectively) in cells from the shearable region as well as protein translation and modification and enhanced expression of protein involved in heat shock response and chaperonin function (DnaK) in cells from the nonshearable region revealed that the physiological status of cells in the two biofilm regions was distinct. This was also reflected in the different morphologies of cells from the two biofilm zones. Stratified patterns of cell metabolism and morphology in biofilms, obtained using shear-induced biofilm fractionation, may yield important information of how cells of deeply embedded biofilm bacteria survive imposed conditions of stress such as treatment with antimicrobial agents or antibiotics.

scholarly journals Role of aspartate ammonia-lyase in Pasteurella multocida

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Zui Wang ◽  
Li Li ◽  
Peng Liu ◽  
Chen Wang ◽  
Qin Lu ◽  
...  
Keyword(s):  
Mutant Strain ◽  
Bacterial Growth ◽  
Type Strain ◽  
Pasteurella Multocida ◽  
Wild Type Strain ◽  
Luria Bertani ◽  
Economic Losses ◽  
Wild Type ◽  
Mueller Hinton ◽  
Significant Difference

Abstract Background Pasteurella multocida is responsible for a highly infectious and contagious disease in birds, leading to heavy economic losses in the chicken industry. However, the pathogenesis of this disease is poorly understood. We recently identified an aspartate ammonia-lyase (aspA) in P. multocida that was significantly upregulated under iron-restricted conditions, the protein of which could effectively protect chicken flocks against P. multocida. However, the functions of this gene remain unclear. In the present study, we constructed aspA mutant strain △aspA::kan and complementary strain C△aspA::kan to investigate the function of aspA in detail. Result Deletion of the aspA gene in P. multocida resulted in a significant reduction in bacterial growth in LB (Luria-Bertani) and MH (Mueller-Hinton) media, which was rescued by supplementation with 20 mM fumarate. The mutant strain △aspA::kan showed significantly growth defects in anaerobic conditions and acid medium, compared with the wild-type strain. Moreover, growth of △aspA::kan was more seriously impaired than that of the wild-type strain under iron-restricted conditions, and this growth recovered after supplementation with iron ions. AspA transcription was negatively regulated by iron conditions, as demonstrated by quantitative reverse transcription-polymerase chain reaction. Although competitive index assay showed the wild-type strain outcompetes the aspA mutant strain and △aspA::kan was significantly more efficient at producing biofilms than the wild-type strain, there was no significant difference in virulence between the mutant and the wild-type strains. Conclusion These results demonstrate that aspA is required for bacterial growth in complex medium, and under anaerobic, acid, and iron-limited conditions.

scholarly journals Dimethyl Adenosine Transferase (KsgA) Deficiency in Salmonella enterica Serovar Enteritidis Confers Susceptibility to High Osmolarity and Virulence Attenuation in Chickens

2013 ◽  
Vol 79 (24) ◽  
pp. 7857-7866 ◽  
Author(s):  
Kim Lam Chiok ◽  
Tarek Addwebi ◽  
Jean Guard ◽  
Devendra H. Shah
Keyword(s):  
Salmonella Enterica ◽  
Cold Sensitivity ◽  
Growth Defect ◽  
Phenotype Microarray ◽  
Wild Type ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Content Type ◽  
High Osmolarity ◽  
Dna Mismatch ◽  
Virulence Attenuation

ABSTRACTDimethyl adenosine transferase (KsgA) performs diverse roles in bacteria, including ribosomal maturation and DNA mismatch repair, and synthesis of KsgA is responsive to antibiotics and cold temperature. We previously showed that aksgAmutation inSalmonella entericaserovar Enteritidis results in impaired invasiveness in human and avian epithelial cells. In this study, we tested the virulence of aksgAmutant (theksgA::Tn5mutant) ofS. Enteritidis in orally challenged 1-day-old chickens. TheksgA::Tn5mutant showed significantly reduced intestinal colonization and organ invasiveness in chickens compared to those of the wild-type (WT) parent. Phenotype microarray (PM) was employed to compare theksgA::Tn5mutant and its isogenic wild-type strain for 920 phenotypes at 28°C, 37°C, and 42°C. At chicken body temperature (42°C), theksgA::Tn5mutant showed significantly reduced respiratory activity with respect to a number of carbon, nitrogen, phosphate, sulfur, and peptide nitrogen nutrients. The greatest differences were observed in the osmolyte panel at concentrations of ≥6% NaCl at 37°C and 42°C. In contrast, no major differences were observed at 28°C. In independent growth assays, theksgA::Tn5mutant displayed a severe growth defect in high-osmolarity (6.5% NaCl) conditions in nutrient-rich (LB) and nutrient-limiting (M9 minimum salts) media at 42°C. Moreover, theksgA::Tn5mutant showed significantly reduced tolerance to oxidative stress, but its survival within macrophages was not impaired. UnlikeEscherichia coli, theksgA::Tn5mutant did not display a cold-sensitivity phenotype; however, it showed resistance to kasugamycin and increased susceptibility to chloramphenicol. To the best of our knowledge, this is the first report showing the role ofksgAinS. Enteritidis virulence in chickens, tolerance to high osmolarity, and altered susceptibility to kasugamycin and chloramphenicol.

Reduction of Salmonella enterica Serovar Enteritidis on the Surface of Raw Shelled Almonds by Exposure to Steam

2006 ◽  
Vol 69 (3) ◽  
pp. 591-595 ◽  
Author(s):  
SUN-YOUNG LEE ◽  
SE-WOOK OH ◽  
HYUN-JUNG CHUNG ◽  
JOSE I. REYES-DE-CORCUERA ◽  
JOSEPH R. POWERS ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enteritidis ◽  
Treatment Time ◽  
Steam Treatment ◽  
Population Estimates ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Varietal Differences ◽  
Significant Difference ◽  
Sublethal Injury ◽  
Over Time

This study was conducted to investigate the effect of steam treatment on the reduction of Salmonella enterica serovar Enteritidis on the surface of raw almonds. Two cultivars, ‘Nonpareil’ and ‘Mission’, were studied. Salmonella Enteritidis was inoculated on the surface of raw almonds, which were then treated with steam (93°C ± 1°C) for 5, 15, 25, 35, 45, 55, and 65 s. After steam treatment, samples were plated on xylose lysine desoxycholate (XLD) and overlay (OV) XLD as a selective and nonselective agar for Salmonella, respectively, to investigate the extent of sublethal injury in Salmonella. Steam treatment of raw almonds effectively reduced Salmonella Enteritidis, and the effect was pronounced with increasing treatment time. After 65 s of steam treatment, reductions in Salmonella Enteritidis populations were 5.7 log and 5.8 log for ‘Nonpareil’ and 4.0 log and 4.1 log for ‘Mission’ when enumerated on XLD and OV XLD, respectively. There was no significant difference in population estimates determined with XLD and OV XLD over time (P &gt; 0.05). The effect of the steam treatment was significantly different between two almond cultivars. Salmonella inoculated onto ‘Mission’ was more resistant to the steam treatment than that on ‘Nonpareil’, indicating that varietal differences must be considered in the application of steam for the disinfection of raw almonds. The present investigation revealed the potential usefulness of steam treatments for the control of pathogens in raw almonds.

scholarly journals Meat juice contributes to the stability of ethanol adaptation in Salmonella enterica serovar Enteritidis

2021 ◽  
Vol 5 ◽  
Author(s):  
Shoukui He ◽  
Karen Fong ◽  
Siyun Wang ◽  
Xianming Shi
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enteritidis ◽  
Ethanol Tolerance ◽  
Luria Bertani ◽  
Ethanol Stress ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Meat Juice ◽  
Luria Bertani Broth ◽  
Food Borne ◽  
The Stability

Abstract Stability assessment of observed tolerance phenotypes is integral in understanding stress adaptation in food-borne pathogens. Therefore, the current work was carried out to determine whether ethanol adaptation induced by exposure to 5 per cent ethanol for 60 min is a stable phenomenon in Salmonella enterica serovar Enteritidis. The capacity of Salmonella Enteritidis (S. Enteritidis) to maintain the acquired ethanol adaptation in the absence of sublethal ethanol stress was investigated at 37 °C, 25 °C or 4 °C in Luria–Bertani broth and two types of meat juice. It was found that ethanol adaptation was completely reversed within 40 min at 37 °C or within 60 min at 25 °C, but was stable at 4 °C for at least 48 h in the broth assay. Ethanol adaptation was retained in chicken juice during 60-min incubation at 25 °C or 48-h incubation at 4 °C. Moreover, exposure to pork juice stored at either 25 °C or 4 °C significantly (P&lt;0.05) increased the ethanol tolerance of ethanol-adapted cells. Collectively, these findings suggest that ethanol adaptation stability in S. Enteritidis under cold conditions and in meat juices should be taken into account when conducting a comprehensive risk analysis during food processing.

scholarly journals The Homolog of the Gene bstA of the BTP1 Phage from Salmonella enterica Serovar Typhimurium ST313 Is an Antivirulence Gene in Salmonella enterica Serovar Dublin

2017 ◽  
Vol 86 (1) ◽  
Author(s):  
Ana Herrero-Fresno ◽  
Irene Cartas Espinel ◽  
Malene Roed Spiegelhauer ◽  
Priscila Regina Guerra ◽  
Karsten Wiber Andersen ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Type Strain ◽  
Wild Type Strain ◽  
Luria Bertani ◽  
Wild Type ◽  
Content Type ◽  
Serovar Typhimurium ◽  
Cattle Blood

ABSTRACTIn a previous study, a novel virulence gene,bstA, identified in aSalmonella entericaserovar Typhimurium sequence type 313 (ST313) strain was found to be conserved in all publishedSalmonella entericaserovar Dublin genomes. In order to analyze the role of this gene in the host-pathogen interaction inS. Dublin, a mutant where this gene was deleted (S. Dublin ΔbstA) and a mutant which was further genetically complemented withbstA(S. Dublin 3246-C) were constructed and tested in models ofin vitroandin vivoinfection as well as during growth competition assays in M9 medium, Luria-Bertani broth, and cattle blood. In contrast to the results obtained for a strain ofS. Typhimurium ST313, the lack ofbstAwas found to be associated with increased virulence inS. Dublin. Thus,S. Dublin ΔbstAshowed higher levels of uptake than the wild-type strain during infection of mouse and cattle macrophages and higher net replication within human THP-1 cells. Furthermore, during mouse infections,S. Dublin ΔbstAwas more virulent than the wild type following a single intraperitoneal infection and showed an increased competitive index during competitive infection assays. Deletion ofbstAdid not affect either the amount of cytokines released by THP-1 macrophages or the cytotoxicity toward these cells. The histology of the livers and spleens of mice infected with the wild-type strain and theS. Dublin ΔbstAmutant revealed similar levels of inflammation between the two groups. The gene was not important for adherence to or invasion of human epithelial cells and did not influence bacterial growth in rich medium, minimal medium, or cattle blood. In conclusion, a lack ofbstAaffects the pathogenicity ofS. Dublin by decreasing its virulence. Therefore, it might be regarded as an antivirulence gene in this serovar.

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