salmonella enterica treatment

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Infection is diagnosed when a laboratory test detects Salmonella bacteria in stool, body tissue, or fluids. In some cases, the diarrhea associated with salmonella infection can be so dehydrating as to require prompt medical attention. This reduction in membrane fluidity correlated with the cellular heat shock response, as detected by the change in the induction levels of GroEL and DnaK (40, 43). In addition to the differential expression of proteins following TSP or alkaline pH treatment, changes in membrane fatty acid composition were also observed. Each year, ≈11–21 million cases and 128,000–161,000 typhoid-related deaths occur, making typhoid a continued health concern in many low- and middle-income countries, particularly among populations without access to clean water or improved sanitation ( 1 ). The U.S. Department of Agriculture has approved the use of trisodium phosphate (TSP) as a postchill processing aid to reduce bacteria on raw poultry carcasses (5, 23). In Pediococcus sp., a decrease in the unsaturated to saturated and cyclic fatty acid ratio during transformation from log to stationary phase was accompanied by an increase in thermotolerance (4). A randomized controlled longitudinal field trial was designed to determine the long-term effects of a single dose of ceftiofur or tulathromycin on Salmonella population characteristics in cattle feces and peripheral … These results suggest that serovar Enteritidis cells would respond in a similar fashion when exposed to a sublethal concentration of TSP or its equivalent alkaline pH. The samples were then desalted and concentrated with ZipTips C18 (Millipore Corp., Bedford, Mass.) %%EOF In 1998 to 2002, empirical treatment for childhood sepsis consists of chloramphenicol. Prescribed antibiotics were third generation cephalosporins and fluoroquinolones. Although alkaline adaptation increases thermotolerance in E. coli and salmonellas, it has not been correlated with the expression of heat shock proteins (HSPs) (30). Influence of TSP or alkaline pH pretreatment on thermotolerance and sensitivity to other stresses. Severe cases may require hospitalization and fluids delivered directly into a vein (intravenous). Histori… A spondylitis and a purulent pleurisy were observed and deemed related to the UTI. Mass spectrometric analysis showed the up-regulated proteins to include those involved in the transport of small hydrophilic molecules across the cytoplasmic membrane and those that act as chaperones and aid in the export of newly synthesized proteins by keeping them in open conformation. At an acidic external pH, amino acid decarboxylases generate amines, which are exported and thereby reverse acidification (22, 41, 45, 46, 69). Acids and bases have been shown to have multiple effects on gene expression in E. coli (for a review, see references 18, 48, and 63). Densitometric analysis of these differentially expressed proteins revealed at least a 1.5-fold increase in expression compared to the untreated control. Two-dimensional polyacrylamide gel electrophoresis (PAGE) of total cellular proteins from untreated control cells resolved as many as 232 proteins, of which 22 and 15% were absent in TSP- or alkaline pH-pretreated cells, respectively. This led to the hypothesis that thermotolerance induced by TSP or alkaline pH pretreatment could involve other stress proteins and/or could be due to some physiological changes that occur during TSP or alkaline pH pretreatment. (16) and Portnoy et al. endstream endobj 135 0 obj <> endobj 136 0 obj <> endobj 137 0 obj <>stream Although these previous studies have correlated similar changes in the composition of the membrane fatty acids during heat shock or acid adaptation to the induction of thermotolerance, this is the first report, to our knowledge, linking thermotolerance induced during alkaline adaptation to changes in membrane fatty acid composition. However, the resistance to pH 11.0 was independent of de novo protein synthesis as no significant (P < 0.05) difference in resistance was observed (Table 1) even when protein synthesis was inhibited with chloramphenicol during TSP or alkaline pH pretreatment. Isoelectric focusing was achieved when the total running time yielded 60 kVh at 20°C. Worldwide there are 16 million annual cases of typhoid fever, 1.3 billion cases of … This question is for testing whether or not you are a human visitor and to prevent automated spam submissions. Pretreatment with 1.5% TSP or pH 10.0 significantly (P < 0.05) induced thermotolerance and resistance to TSP and high pH while sensitizing serovar Enteritidis cells to acid and H2O2 (Table 1). While some of the infections can be easily treated, some of the strains have been shown to resist antibiotic treatment and are therefore deadly. Bacteria and culture conditions. 134 0 obj <> endobj These results suggest that the cytoplasmic membrane could play a significant role in the induction of thermotolerance and resistance to other stresses following TSP or alkaline pH treatment. Adaptation of Salmonella or E. coli to alkaline conditions is accompanied by induced thermotolerance, increased resistance to bile salts, and increased resistance to high pH (17, 24, 32, 71). The responses of Salmonella enterica serovar Enteritidis to a sublethal dose of trisodium phosphate (TSP) and its equivalent alkaline pH made with NaOH were examined. Salmonella enterica serovar Typhi is a gram-negative, rod-shaped facultative anaerobe that only infects humans. Table 1 shows the D values of 1.5% TSP- or pH 10.0-pretreated cells of S. enterica serovar Enteritidis challenged with either 55°C, 2.5% TSP, pH 11.0 (pH equivalent to a treatment solution containing 2.5% TSP), pH 2.6, or 30 mM H2O2. The gels were scanned as mentioned earlier, and densitometric analysis of differentially expressed proteins was performed with an IS-1000 Digital Imaging System (Alpha Innotech Corp., San Leandro, Calif.). The responses of Salmonella enterica serovar Enteritidis to a sublethal dose of trisodium phosphate (TSP) and its equivalent alkaline pH made with NaOH were examined. One of the most important consequences of membrane fatty acid changes in microorganisms is modulation of the activity of intrinsic proteins that perform functions such as ion pumping and nutrient uptake (57). While this provides a possible explanation for S. enterica serovar Enteritidis survival in 1.5% TSP or pH 10.0 treatment solutions, pretreatment with TSP or alkaline pH resulted in the down-regulation of a translational elongation factor (Table 2) involved in the binding of aminoacyl-tRNA to the A site of ribosomes during protein synthesis. 1C) or down-regulated (Fig. ��^`�� U �ȰǀS����[�A�P����l'�L//��9s�ݿ0�;R��s�*�_v,xA�m%�{��5n_u���h+�{��n_�y!���FY�x�CI���9N��������CF��� N�=[��FY�ݳ]������M>mͿs���-�s�5~7���C�� @K���L�wt40T�ht�(�Ё���ʠ��@�a��@�H ۋA�g:�bypؤ13p?�o`�e0��b�� ^�,����8�A'�%�����|�L�t4�Uf~�Ր� �� �����1k��uL)��a7� 1989年以降に鶏卵汚染のSalmonella(S.) Enteritidisが急増し、現在もその傾向は続いています。 この細菌は家畜や野生動物、爬虫類等に保菌されていることが多く、下水、河川や沼にも生息しています。したがって動物の糞便や汚染水等からさまざまな食材を汚染し、洗浄の不完全、加熱不足や他への二次汚染などが原因となり食中毒が発生します。 畑作栽培では土壌や水からの汚染が避けられませんし、水耕栽培するスプラウト(発芽野菜)では汚染された種子や、栄養として与える使用水の汚染が原因とな … The FAME peaks were automatically integrated by Hewlett Packard 3365 ChemStation software, and FAMEs were identified with the MIDI Microbial Identification Software (Sherlock TSBA Library version 3.80; Microbial ID, Inc., Newark, Del.). For an infection to occur, the intracellular growth of Salmonella in macrophages is crucial. Infection usually causes relatively mild symptoms but complications, including lack of fluid in the body (dehydration), can occur in some cases. Antibiotic use in cattle can select for multidrug-resistant Salmonella enterica, which is considered a serious threat by the U.S. Centers for Disease Control and Prevention. The induction of thermotolerance following alkaline adaptation has previously been shown to be dependent on protein synthesis (33), whereas resistance to high pH appeared to be due to a phenotypic change (25, 32) during alkaline adaptation. Protein inhibition studies with chloramphenicol revealed that thermotolerance, unlike resistance to high pH, was dependent on de novo protein synthesis. The enterica subspecies of Salmonella enterica has at least 2,610 different serotypes. Among more than 2,000 serotypes, Salmonella enterica serotype Choleraesuis shows the highest predilection to cause systemic infections in humans. D values of pretreated S. enterica serovar Enteritidis ATCC 4931 challenged with different lethal stresses. Treatment of inoculated hides with 1,000 ppm SDIC for 5 min resulted in reduction of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes at … The observation of up-regulation of proteins involved in amino acid biosynthesis during TSP or alkaline pH treatment is in agreement with observations of Slonczewski and coworkers (7, 68). Only protein spots that appeared in four out of six gels were selected for analysis. 1A), respectively, by pH 10.0 pretreatment. Cell pellets were resuspended in 0.5 ml of 50 mM HEPES buffer (pH 7.4) and, while kept on ice, sonicated at the same settings as mentioned previously. TFA (0.1%) was then aspirated five times through the ZipTips to remove salts, and the sample was then eluted directly onto the MALDI plate with 75% acetonitrile containing 5 mg of α-cyano-4-hydroxycinnamic acid per ml. Newport has been ranked among the top five serovars responsible for persistent infections, globally. The organism was isolated from the sputum, the protected specimen brush material of bronchial secretions, and the stool. Growth conditions such as the composition of the growth medium (2, 3, 35, 60), the growth phase (age) of the cells (35, 42, 50, 75), the incubation temperature at which the bacteria were cultured (2, 35, 55, 56, 60, 70), and the pH (10, 56, 60) markedly affect the composition of the membrane lipid. This would mean that protein synthesis in TSP- or alkaline pH-pretreated cells would be inhibited compared to that of untreated controls, thus explaining the observed reduction in the number of proteins expressed during these pretreatments. In the control treatment from the previous study (59), stationary-phase cells were held in fresh TSB for 1 h and no significant increase in cell numbers was observed. Levels of multidrug resistance have become alarmingly high. Differentially expressed proteins were then detected and quantified from the stored images with PDQuest 2-D Analysis Software (Bio-Rad). The sonicated cells were then centrifuged at 16,000 × g for 2 min at 4°C to remove cell debris. The prevalence of Salmonella enterica serotype Paratyphi A infection is increasing, and multidrug resistance is a well-recognized problem. 0 1A), 22% were not present in 1.5% TSP-treated cells (Fig. Conversely, alkaline adaptation of S. enterica serovar Enteritidis or E. coli sensitizes the cells to acid stress (51) and vice versa (53). The cytoplasmic membrane, being the boundary between the cytoplasm and the external environment, is the first site in the bacterial cell that is exposed to various stresses. However, the relationship between alkaline stress and the heat shock response is not very clear. Pretreatment of S. enterica serovar Enteritidis cells with 1.5% TSP or pH 10.0 solutions resulted in a significant increase in thermotolerance, resistance to 2.5% TSP, resistance to high pH, and sensitivity to acid and H 2 O 2. Differential expression of proteins by E. coli and salmonellas due to changes in their growth environment (nutrient-limiting conditions, anaerobiosis) (7, 67) or exposure to various stresses has been reported by a number of workers (1, 8, 20, 30, 39, 67). 1B) or down-regulated (Fig. Following isoelectric focusing, the gels were equilibrated twice for 10 min each time in fresh isoelectric focusing gel equilibration buffer (50 mM Tris-HCl [pH 8.8], 6 M urea, 30% [vol/vol] glycerol, 2.0% [wt/vol] SDS, 65 mM DTT, 0.01% [wt/vol] bromophenol blue). 2D gels for each treatment were run in duplicate three times to confirm the reproducibility of the protein pattern. Salmonella infection is so common, however, that drug-resistant strains are prevalent. In 2005, Salmonella enterica finally gained. Salmonella enterica serovars Typhi, Paratyphi A, and Paratyphi B are the etiologic agents of typhoid and paratyphoid fever. NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. The sublethal dose of TSP used was determined from preliminary experiments. Introduction. For adults who did not respond to initial treatments or were unable to take oral medication, they were prescribed parenteral gentamicin. Analysis of protein spots on 2D gels.Individual gels were scanned on an Epson 1200C scanner with a transparency adapter as 8-bit grayscale 300-dot/in. Cells in the stationary phase of growth were prepared as described previously (59) and used for various treatments. prolonged antibiotic treatment remains a major therapeutic option. A study of data from 37 countries collected between 2001 and 2007 found that the most common serovar of Salmonella isolated from human cases was Enteritidis, found in an average of 43.5% of cases, followed by Typhimurium (17.1% of cases), Newport (3.5%), Infantis (1.8%), Virchow (1.5%), Hadar (1.5%), and Agona (0.8%). The SDS-PAGE profile of these proteins showed differential expression of as many as four proteins compared to the untreated controls (Fig. The MALDI-time of flight mass spectrometer (Voyager DE-STR; Applied Biosystems, Foster City, Calif.) was operated in the positive-ion reflectron mode. RpoS is an alternative sigma factor of bacterial RNA polymerase (34, 72) that mediates the increase in the expression of many genes during the transition to stationary phase (29). The precipitate was collected by centrifugation at 4,500 × g for 15 min (Eppendorf 5804R benchtop centrifuge), resuspended in 2.0 ml of protein-solubilizing solution (9.0 M urea, 4.0% [wt/vol] CHAPS, 100 mM DTT, 2% [vol/vol] pH 4.0 to 7.0 IPG buffer), and held at RT for 1 h. The dye-binding assay of Bradford was then performed to quantify the protein concentration (9). There are two major species that include: 1. Lane 1 is a prestained protein molecular mass ladder. Analysis of total cellular proteins by 1D SDS-PAGE and immunoblotting of total cellular proteins.Total cellular protein extracted by sonication (10 bursts, 10 s each at 60 W, Sonifier Cell Disruptor; Branson Ultrasonic Corp., Danbury, Conn.) of the cells in 50 mM HEPES (pH 7.4) were separated by one-dimensional (1D) SDS-polyacrylamide gel electrophoresis (PAGE) (38). After prolonged antimicrobial chemotherapy and TSP treatment has been shown to significantly reduce the populations of Escherichia coli O157:H7 (15, 73), Salmonella enterica serovar Enteritidis (27), S. enterica serovar Typhimurium (15, 74), Clostridium sporogenes (14, 15), Campylobacter jejuni (58, 66), and Listeria monocytogenes (13, 58, 66) on animal carcasses. ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology. Journal of Microbiology & Biology Education, Microbiology and Molecular Biology Reviews. Treatment of inoculated hides with 1,000 ppm SDIC for 5 min resulted in reduction of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes at … The samples were then aspirated and dispensed five times through the ZipTips to bind the peptides to the ZipTips. The most feared complication of serotype Cholearesuis bacteremia in adults is the development of mycotic aneurysm, which previously was … Our findings highlight the utility of enhanced genomic surveillance for typhoid fever in this region. The concentration of each lethal factor was determined by preliminary experiments in which several concentrations of each lethal factor were tested. The average treatment duration was 20 days. Our results also confirm that the induction of thermotolerance, unlike the resistance to high pH, was dependent on protein synthesis (Table 1). More than 50% of the proteins that were either up- or down-regulated by TSP pretreatment were also up- or down-regulated by alkaline pH pretreatment. In this study stationary-phase cells were used to examine the effect of pretreatment of S. enterica serovar Enteritidis with a sublethal concentration of TSP or alkaline pH on tolerance to lethal stresses, protein synthesis, and membrane fatty acid composition. Cause of Typhi salmonellosis (Typhi fever, paraTyphi A, B and C), Salmonella enterica serotype typhi or paratyphi A, B and C include salmonella to the family of Enterobacteriaceae. h�bbd```b``�"�@$�Q��Q �,�A��l��K`2,^f��ٻ�� `�6�d�"�[A��^�ii �$ ���1�`�g���S������������ ��a In addition to the role of HSPs in the induction of thermotolerance, modifications in membrane fluidity have also been shown to be involved in the adaptation to thermal stress. tory of systemic lupus erythematosus and had been treated with prednisolone and azathioprine. a decrease in the unsaturated to saturated and cyclic fatty acid ratio during the transformation from log- to stationary-phase growth was accompanied by an increase in the D values, suggesting a relationship between thermotolerance and reduced membrane fluidity (4). The invasiveness of Salmonella enterica serovar Typhimurium (S.Typhimurium) is closely associated with the Salmonella pathogenicity island (SPI)‐encoded type Ⅲ secretion system (T3SS), which can directly inject a series of effector proteins into eukaryotic cells to enable bacterial infection. Influence of TSP or alkaline pH pretreatment on thermotolerance and sensitivity to other stresses.In a previous study to elucidate the mechanism of antimicrobial activity of TSP we reported that stationary-phase cells of S. enterica serovar Enteritidis ATCC 4931 can survive exposure to 1.5% TSP or its equivalent alkaline pH of 10.0, made with NaOH, for 1 h (59). ( TFA ) serotypes, Salmonella species have the serologically defined names appended as serovars or serotypes cramps within to!, Paratyphi a infection is so common, however, to our knowledge, there are two major species include! 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The infection included common housekeeping proteins like those involved in amino acid catabolism are induced min... Introductions of Salmonella in macrophages is crucial Africa and associated with Salmonella infection have no symptoms introduced from Europe multiple! Va. ) to ampicillin and chloramphenicol caused epidemics in Latin America may eat than fourfold patients... Not clear and does not seem to involve the induction of HSPs but to!

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